1 00:00:05,680 --> 00:00:09,960 >>WELCOME TO NIH WEDNESDAY 2 00:00:09,960 --> 00:00:25,800 AFTERNOON LECTURE SERIES 3 00:00:25,800 --> 00:00:27,400 IT'S REALLY A GREAT PLEASURE TO 4 00:00:27,400 --> 00:00:30,880 INTRODUCE DR. ANDREA CALIFANO TO 5 00:00:30,880 --> 00:00:31,640 YOU. 6 00:00:31,640 --> 00:00:33,760 I'VE KNOWN ANDREA BECAUSE OF A 7 00:00:33,760 --> 00:00:38,160 JOINT INTEREST IN BOTH GENOMICS 8 00:00:38,160 --> 00:00:42,160 AND EARLY ON HIS INTEREST IN D 9 00:00:42,160 --> 00:00:43,440 CELL LYMPHOMAS, NOW INTERESTED 10 00:00:43,440 --> 00:00:45,200 IN ALL CANCER, AND HE'S GOING TO 11 00:00:45,200 --> 00:00:50,440 BE DISCUSSING SYSTEMS BIOLOGY 12 00:00:50,440 --> 00:00:57,320 NETWORK-BASED METHODS FOR 13 00:00:57,320 --> 00:00:58,720 IDENTIFYING THERAPEUTIC TARGETS. 14 00:00:58,720 --> 00:01:03,960 I THINK HIS FOCUS IS TO USE 15 00:01:03,960 --> 00:01:05,040 THESE GENOMIC CYST TIM'S BIOLOGY 16 00:01:05,040 --> 00:01:07,560 WAYS TO ACTUALLY DO NEW CLINICAL 17 00:01:07,560 --> 00:01:08,560 TRIALS WHICH HE'LL TELL US 18 00:01:08,560 --> 00:01:09,640 ABOUT. 19 00:01:09,640 --> 00:01:15,240 OUR SPEAKER IS EXTREMELY 20 00:01:15,240 --> 00:01:16,440 WELL-KNOWN WITHIN THE CANCER 21 00:01:16,440 --> 00:01:18,080 COMMUNITY AND MORE GENERALLY IN 22 00:01:18,080 --> 00:01:19,120 SYSTEMS BIOLOGY COMMUNITY. 23 00:01:19,120 --> 00:01:23,200 HE IS THE CLYDE AND HELEN WU 24 00:01:23,200 --> 00:01:24,280 PROFESSOR OF CHEMICAL SYSTEMS 25 00:01:24,280 --> 00:01:25,280 BIOLOGY, CHAIR OF THE DEPARTMENT 26 00:01:25,280 --> 00:01:30,800 OF SYSTEMS BIOLOGY AND DIRECTOR 27 00:01:30,800 --> 00:01:37,320 OF THE J.P.SULZBERGER COLUMBIA 28 00:01:37,320 --> 00:01:38,600 GENOME CENTER AT THE COLUMBIA 29 00:01:38,600 --> 00:01:39,600 UNIVERSITY MEDICAL CENTER. 30 00:01:39,600 --> 00:01:40,560 ORIGINALLY A PHYSICIST BY 31 00:01:40,560 --> 00:01:43,920 TRAINING AND HE'S TAKEN IN MY 32 00:01:43,920 --> 00:01:46,280 VIEW QUITE INNOVATIVE APPROACHES 33 00:01:46,280 --> 00:01:51,800 TO IDENTIFYING LIKELY FACTORS 34 00:01:51,800 --> 00:01:53,600 INVOLVED IN CANCER PROGRESSION, 35 00:01:53,600 --> 00:01:54,720 CANCER PHENOTYPES IN GENERAL, 36 00:01:54,720 --> 00:01:56,400 DRUG RESPONSE AND RESISTANCE, 37 00:01:56,400 --> 00:01:58,160 AND MORE RECENTLY I THINK AS 38 00:01:58,160 --> 00:02:03,280 HE'LL TELL YOU ABOUT TODAY, 39 00:02:03,280 --> 00:02:04,760 USING SINGLE CELL DATA FROM 40 00:02:04,760 --> 00:02:06,240 CANCER PATIENTS TO PREDICT 41 00:02:06,240 --> 00:02:09,400 PATHWAY ACTIVATION. 42 00:02:09,400 --> 00:02:11,800 THIS INVOLVES A LOT OF MATH, 43 00:02:11,800 --> 00:02:12,880 WHICH ANDREA IS VERY GOOD AT, 44 00:02:12,880 --> 00:02:16,000 AND IT INVOLVES INTEGRATING INTO 45 00:02:16,000 --> 00:02:21,600 NETWORK MODELS TO MAKE A TOTAL 46 00:02:21,600 --> 00:02:22,840 MODEL OF THE BEHAVIOR OF AT 47 00:02:22,840 --> 00:02:28,720 LEAST PARTS OF THE CANCER CELL. 48 00:02:28,720 --> 00:02:31,200 HE'S SHOWN I THINK HIS MAJOR 49 00:02:31,200 --> 00:02:32,200 CONTRIBUTION IN MY VIEW TO 50 00:02:32,200 --> 00:02:34,000 CANCER BIOLOGY LITERATURE AS THE 51 00:02:34,000 --> 00:02:36,880 IDEA OF THE MASTER REGULATOR. 52 00:02:36,880 --> 00:02:38,560 WHICH IS BASICALLY THESE ARE 53 00:02:38,560 --> 00:02:40,720 NODES WITH -- REGULATORY NODES 54 00:02:40,720 --> 00:02:44,000 WITHIN CANCER CELLS THAT HE CAN 55 00:02:44,000 --> 00:02:48,400 IDENTIFY COMPUTATIONALLY THAT 56 00:02:48,400 --> 00:02:49,240 INTEGRATE A LOT OF SIGNALS AND 57 00:02:49,240 --> 00:02:53,200 THEN HAVE OUTPUT TO MANY 58 00:02:53,200 --> 00:02:55,320 DOWNSTREAM TARGETS AND THAT 59 00:02:55,320 --> 00:02:56,880 THESE ARE CALLED MASTER 60 00:02:56,880 --> 00:02:59,400 REGULATORS THAT IF YOU INTERFERE 61 00:02:59,400 --> 00:03:02,760 WITH THESE, THEN THEY HAVE SUCH 62 00:03:02,760 --> 00:03:04,200 A BROAD EFFECT ON THE CANCER 63 00:03:04,200 --> 00:03:07,240 CELL THAT TYPICALLY THAT INDUCES 64 00:03:07,240 --> 00:03:08,760 CELL DEATH OR A MAJOR CHANGE IN 65 00:03:08,760 --> 00:03:11,920 PHENOTYPE. 66 00:03:11,920 --> 00:03:12,920 AND THIS THEN LEADS OBVIOUSLY TO 67 00:03:12,920 --> 00:03:16,640 A NEW SET OF THERAPEUTIC TARGETS 68 00:03:16,640 --> 00:03:23,760 IN CANCER THAT CAN BE ADDRESSED 69 00:03:23,760 --> 00:03:25,720 BY A NUMBER OF MEANS, 70 00:03:25,720 --> 00:03:26,120 PHARMACOLOGICALLY. 71 00:03:26,120 --> 00:03:28,040 MANY OF THESE ARE TRANSCRIPTION 72 00:03:28,040 --> 00:03:30,040 FACTORS SO THEY MAY REQUIRE 73 00:03:30,040 --> 00:03:33,400 THINGS LIKE PROTACS TO BE DONE 74 00:03:33,400 --> 00:03:34,120 TO BE TARGETED. 75 00:03:34,120 --> 00:03:35,800 SO JUST A LITTLE BIT MORE 76 00:03:35,800 --> 00:03:37,800 DETAIL, HIS UNDERGRADUATE DEGREE 77 00:03:37,800 --> 00:03:39,920 AND PH.D. WAS IN PHYSICS AT THE 78 00:03:39,920 --> 00:03:41,640 UNIVERSITY OF FLORENCE IN ITALY, 79 00:03:41,640 --> 00:03:44,800 HIS PH.D. WAS ON THE STUDY OF 80 00:03:44,800 --> 00:03:48,800 CHAOTIC BEHAVIOR IN HIGH 81 00:03:48,800 --> 00:03:50,200 DIMENSIONAL SYSTEMS, HAVING READ 82 00:03:50,200 --> 00:03:53,000 THE BOOK "CHAOS," THIS DOES NOT 83 00:03:53,000 --> 00:03:56,000 MEAN THAT HIS DISSERTATION WAS 84 00:03:56,000 --> 00:03:57,240 CHAOTIC, BUT IT WAS A TYPE OF 85 00:03:57,240 --> 00:03:59,320 BROAD ANALYSIS OF COMPLEX 86 00:03:59,320 --> 00:04:02,080 BEHAVIOR OF DYNAMICAL SYSTEMS 87 00:04:02,080 --> 00:04:03,240 THAT I THINK IS A GOOD WAY TO 88 00:04:03,240 --> 00:04:06,400 THINK ABOUT HOW BIOLOGICAL 89 00:04:06,400 --> 00:04:10,840 SYSTEMS MIGHT ACTUALLY WORK. 90 00:04:10,840 --> 00:04:13,200 HE THEN MOVED ON TO POSTDOCTORAL 91 00:04:13,200 --> 00:04:15,240 TRAINING IN FLORENCE, MIT AND 92 00:04:15,240 --> 00:04:16,480 FINALLY SPENT QUITE SOME YEARS 93 00:04:16,480 --> 00:04:19,640 AT THE IBM WATSON RESEARCH 94 00:04:19,640 --> 00:04:21,800 CENTER, IN THE 1980s, BECOMING 95 00:04:21,800 --> 00:04:26,840 A STAFF MEMBER THERE, AND THEN 96 00:04:26,840 --> 00:04:31,520 MOVED ON TO COLUMBIA BACK INTO 97 00:04:31,520 --> 00:04:32,800 FULL ACADEMIA, SHALL WE SAY, 98 00:04:32,800 --> 00:04:36,320 ALTHOUGH I THINK HE WAS DOING 99 00:04:36,320 --> 00:04:39,440 ACADEMICS AT THE WATSON RESEARCH 100 00:04:39,440 --> 00:04:40,000 CENTER. 101 00:04:40,000 --> 00:04:42,280 HE MOVED IN 2003 AND EVENTUALLY 102 00:04:42,280 --> 00:04:48,760 BECAME PROFESSOR OF BIOMEDICAL 103 00:04:48,760 --> 00:04:51,040 INFORMATICS, AND NOW AS I 104 00:04:51,040 --> 00:04:55,240 MENTIONED, DIRECTOR OF THE 105 00:04:55,240 --> 00:04:56,120 SALZBERGER COLUMBIA GENOME 106 00:04:56,120 --> 00:04:56,680 CENTER. 107 00:04:56,680 --> 00:04:58,400 HE'S A VERY ENERGETIC GUY IF YOU 108 00:04:58,400 --> 00:05:00,120 GET TO KNOW HIM, AND HE'S 109 00:05:00,120 --> 00:05:01,680 FOUNDED SEVERAL COMPANIES SUCH 110 00:05:01,680 --> 00:05:06,800 AS FIRST GENETIC TRUST AND 111 00:05:06,800 --> 00:05:07,640 THERASIS. 112 00:05:07,640 --> 00:05:09,480 HE'S GOT MANY HONORS, VERY 113 00:05:09,480 --> 00:05:12,200 RIGHTFULLY IN MY OPINION HAS 114 00:05:12,200 --> 00:05:15,360 BEEN AWARDED THE NCI OUTSTANDING 115 00:05:15,360 --> 00:05:16,760 INVESTIGATOR AWARD, HE RECEIVED 116 00:05:16,760 --> 00:05:19,400 THE RUTH SIEGEL AWARD IN 117 00:05:19,400 --> 00:05:20,360 PANCREATIC CANCER RESEARCH, 118 00:05:20,360 --> 00:05:22,400 MEMBER OF THE IBM ACADEMY OF 119 00:05:22,400 --> 00:05:24,480 TECHNOLOGY, AND NOTABLY OF THE 120 00:05:24,480 --> 00:05:26,800 NATIONAL ACADEMY OF MEDICINE. 121 00:05:26,800 --> 00:05:31,600 THE TIE TITLE OF THE TALK IS 122 00:05:31,600 --> 00:05:33,000 ELUCIDATION AND PHARMACOLOGICAL 123 00:05:33,000 --> 00:05:35,880 TARGETING OF NON-ONCOGENE 124 00:05:35,880 --> 00:05:37,520 DEPENDENCIES AT THE SINGLE-CELL 125 00:05:37,520 --> 00:05:37,720 LEVEL. 126 00:05:37,720 --> 00:05:38,480 ANDREA, IT'S A PLEASURE TO HAVE 127 00:05:38,480 --> 00:05:40,040 YOU COME TO THE NIH, AND I'M 128 00:05:40,040 --> 00:05:42,600 ALWAYS EXCITED TO LEARN FROM 129 00:05:42,600 --> 00:05:44,720 YOUR LECTURES, SO WE'RE REALLY 130 00:05:44,720 --> 00:05:46,080 LOOKING FORWARD TO YOUR 131 00:05:46,080 --> 00:05:46,600 PRESENTATION. 132 00:05:46,600 --> 00:05:47,480 THANKS A LOT. 133 00:05:47,480 --> 00:05:49,360 >> THANK YOU FOR THAT WONDERFUL 134 00:05:49,360 --> 00:05:49,680 INTRODUCTION. 135 00:05:49,680 --> 00:05:51,200 I ALSO WANT TO THANK YOU AND 136 00:05:51,200 --> 00:05:52,200 MICHAEL GOTTESMAN FOR INVITING 137 00:05:52,200 --> 00:05:54,680 ME TO GIVE THIS LECTURE. 138 00:05:54,680 --> 00:05:56,960 I THINK THE INVITATION CAME IN 139 00:05:56,960 --> 00:05:58,400 MARCH 2020, AND THEN WE KNOW 140 00:05:58,400 --> 00:06:02,800 WHAT HAPPENED, SO I REALLY AM 141 00:06:02,800 --> 00:06:04,280 GLAD WE'RE FINALLY ABLE TO DO IT 142 00:06:04,280 --> 00:06:05,160 EVEN IF NOT IN PERSON. 143 00:06:05,160 --> 00:06:08,640 THE REASON I HAVE THESE LOGOS 144 00:06:08,640 --> 00:06:11,040 HERE IS BECAUSE MY ENTIRE FIELD 145 00:06:11,040 --> 00:06:16,640 WOULD NOT HAVE DEVELOPED CANCER 146 00:06:16,640 --> 00:06:17,920 SYSTEMS BIOLOGY WITHOUT 147 00:06:17,920 --> 00:06:25,400 TREMENDOUS EFFORTS BY THE NIH 148 00:06:25,400 --> 00:06:27,400 AND NCC AND HOW THE NCI CAN 149 00:06:27,400 --> 00:06:28,520 REALLY FOSTER OF KIND OF STUDIES 150 00:06:28,520 --> 00:06:32,200 THAT I'LL BE DISCUSSING TODAY. 151 00:06:32,200 --> 00:06:35,640 YOU'VE SEEN MY CONFLICT OF 152 00:06:35,640 --> 00:06:36,440 INTEREST ALREADY. 153 00:06:36,440 --> 00:06:38,200 THIS IS THE LATEST COMPANY I 154 00:06:38,200 --> 00:06:41,080 STARTED IN 1916 THAT LICENSED 155 00:06:41,080 --> 00:06:42,840 SOME OF THE ALGORITHMS I'LL BE 156 00:06:42,840 --> 00:06:43,400 DISCUSSING TODAY. 157 00:06:43,400 --> 00:06:44,600 SO IT ALL STARTED FROM WHAT WE 158 00:06:44,600 --> 00:06:46,920 HAVE COME TO CALL THE CANCER 159 00:06:46,920 --> 00:06:51,320 PARADOX. 160 00:06:51,320 --> 00:06:53,400 AND THIS WAS PUBLISHED IN NATURE 161 00:06:53,400 --> 00:06:54,880 REVIEWS, CANCER, AND BASICALLY 162 00:06:54,880 --> 00:06:56,160 THE PARADOX STATES SOMETHING 163 00:06:56,160 --> 00:06:57,520 QUITE OBVIOUS THAT A LOT OF US 164 00:06:57,520 --> 00:07:00,640 HAVE KNOWN FOR AGES, BUT THE 165 00:07:00,640 --> 00:07:01,680 DIFFERENCE HERE THAT WE ACTUALLY 166 00:07:01,680 --> 00:07:04,200 TRY TO FIND POTENTIAL 167 00:07:04,200 --> 00:07:05,080 COMPUTATIONAL SOLUTION TO 168 00:07:05,080 --> 00:07:06,000 ADDRESS THIS PROBLEM. 169 00:07:06,000 --> 00:07:07,600 SO THE PARADOX IS THE FACT THAT 170 00:07:07,600 --> 00:07:08,560 IF YOU LOOK AT THE 171 00:07:08,560 --> 00:07:11,200 TRANSCRIPTIONAL STATE OF CANCER 172 00:07:11,200 --> 00:07:14,320 CELLS, TUMORS BREAK DOWN INTO 173 00:07:14,320 --> 00:07:19,360 SUBTYPES THAT ARE SO TIGHTLY 174 00:07:19,360 --> 00:07:20,800 REGULATED THAT THEY'RE NO MORE 175 00:07:20,800 --> 00:07:21,840 DIFFERENT THAN EQUIVALENT NUMBER 176 00:07:21,840 --> 00:07:23,800 OF SAMPLES TAKEN FROM NORMAL 177 00:07:23,800 --> 00:07:25,400 TISSUES OF THE SAME NUMBER OF 178 00:07:25,400 --> 00:07:25,880 INDIVIDUALS. 179 00:07:25,880 --> 00:07:28,400 SO THE REVIEWERS FROM THAT 180 00:07:28,400 --> 00:07:29,400 PERSPECTIVE DIDN'T BELIEVE THAT 181 00:07:29,400 --> 00:07:30,640 STATEMENT, SO THEY ASKED US TO 182 00:07:30,640 --> 00:07:32,400 GENERATE -- I'M JUST SHOWING YOU 183 00:07:32,400 --> 00:07:35,200 FOUR TUMOR TYPES, AND YOU CAN 184 00:07:35,200 --> 00:07:37,560 SEE OF COURSE BLUE IS TUMOR, RED 185 00:07:37,560 --> 00:07:41,600 IS NORMAL TISSUE, AND YOU CAN 186 00:07:41,600 --> 00:07:43,200 SEE BASICALLY THE DISTRIBUTION 187 00:07:43,200 --> 00:07:47,600 LOOKS VERY SIMILAR. 188 00:07:47,600 --> 00:07:48,280 SO TRANSCRIPTIONALLY, THE CELLS 189 00:07:48,280 --> 00:07:50,040 ARE VERY, VERY SIMILAR. 190 00:07:50,040 --> 00:07:52,560 AND YET AT THE GENETIC LEVEL, 191 00:07:52,560 --> 00:07:54,560 THE MUTATIONS ARE LITERALLY ALL 192 00:07:54,560 --> 00:07:59,840 OVER THE PLACE, SO -- IN TRIPLE 193 00:07:59,840 --> 00:08:01,000 NEGATIVE BREAST CANCER, THERE'S 194 00:08:01,000 --> 00:08:02,000 REALLY NO TWO PATIENTS THAT HAVE 195 00:08:02,000 --> 00:08:04,440 THE SAME SET OF MUTATIONS. 196 00:08:04,440 --> 00:08:05,600 SO WE ARGUED FOR A WHILE ON HOW 197 00:08:05,600 --> 00:08:06,800 WE COULD POTENTIALLY FIND A 198 00:08:06,800 --> 00:08:08,080 SOLUTION TO THIS PROBLEM THAT 199 00:08:08,080 --> 00:08:16,000 WOULD EXPLAIN EX-WHY THAT EXPLAIN WHY THA T IS AND WE 200 00:08:16,000 --> 00:08:18,600 WENT BACK TO -- ONLY ONE 201 00:08:18,600 --> 00:08:19,400 POSSIBLE WAY IN WHICH THIS COULD 202 00:08:19,400 --> 00:08:21,000 BE HAPPENING IS IF ALL THE 203 00:08:21,000 --> 00:08:22,440 MUTATIONS OCCUR UPSTREAM OVER A 204 00:08:22,440 --> 00:08:24,800 PIECE OF MACHINERY THAT IN 205 00:08:24,800 --> 00:08:26,640 CANCER WE CALL THE TUMOR 206 00:08:26,640 --> 00:08:27,720 CHECKPOINT IN FELON CANCER 207 00:08:27,720 --> 00:08:30,640 TISSUE WE CALL THE REGULATORY 208 00:08:30,640 --> 00:08:31,920 CHECKPOINT MODULE, WHERE A VERY 209 00:08:31,920 --> 00:08:35,040 SMALL NUMBER OF PROTEINS WE 210 00:08:35,040 --> 00:08:37,280 DUBBED MASTER REGULATORS, ARE 211 00:08:37,280 --> 00:08:40,160 ACTUALLY INTEGRATING THE 212 00:08:40,160 --> 00:08:42,080 ABERRANT SIGNALS THAT COME FROM 213 00:08:42,080 --> 00:08:46,160 THE MUTATIONS BUT ALSO FROM BOTH 214 00:08:46,160 --> 00:08:49,080 THE DISTAL AND TUMOR ENVIRONMENT 215 00:08:49,080 --> 00:08:49,800 RELATED SIGNALS. 216 00:08:49,800 --> 00:08:51,440 THEN THESE PROTEINS ESSENTIALLY 217 00:08:51,440 --> 00:08:53,800 ARE INCREDIBLY TIGHTLY 218 00:08:53,800 --> 00:08:54,520 OVERREGULATED AND THEREFORE 219 00:08:54,520 --> 00:08:56,400 THEIR STABILITY IS DETERMINED BY 220 00:08:56,400 --> 00:08:58,840 THEIR INTERACTIONS, AND TOGETHER 221 00:08:58,840 --> 00:09:01,040 THEY REGULATE THE 222 00:09:01,040 --> 00:09:01,640 TRANSCRIPTIONAL STATE OF THE 223 00:09:01,640 --> 00:09:02,160 CANCER CELL. 224 00:09:02,160 --> 00:09:03,720 SO THIS PARADIGM WHICH ARE 225 00:09:03,720 --> 00:09:05,800 CALLED AN ONCOTECH TOUR 226 00:09:05,800 --> 00:09:08,200 BASICALLY STATES WHETHER IT'S 227 00:09:08,200 --> 00:09:09,280 REALLY OCCURRING IN CANCER IS 228 00:09:09,280 --> 00:09:10,480 NOT THAT MUCH THE MUTATIONS BUT 229 00:09:10,480 --> 00:09:11,640 RATHER -- THIS IS NOT JUST IN 230 00:09:11,640 --> 00:09:13,800 CANCER BUT ALSO IN COMMON -- BUT 231 00:09:13,800 --> 00:09:15,480 RATHER THE ARCHITECTURE, THE 232 00:09:15,480 --> 00:09:17,520 SIGNALING AND TRANSCRIPTIONAL 233 00:09:17,520 --> 00:09:19,000 ARCHITECTURE THAT IS RESPONSIBLE 234 00:09:19,000 --> 00:09:20,400 FOR THEIR INTEGRATION AND THEIR 235 00:09:20,400 --> 00:09:22,120 TRANSLATION INTO A 236 00:09:22,120 --> 00:09:22,680 TRANSCRIPTIONAL STATE OF THE 237 00:09:22,680 --> 00:09:23,000 CELL. 238 00:09:23,000 --> 00:09:26,040 SO WE HAVE PUBLISHED EXTENSIVELY 239 00:09:26,040 --> 00:09:29,360 ON THESE POINTS AND BASICALLY 240 00:09:29,360 --> 00:09:31,040 USED THE ALGORITHMS TO MAKE 241 00:09:31,040 --> 00:09:31,840 PREDICTION AND GOING TO THE LAB 242 00:09:31,840 --> 00:09:33,400 AND MAKING EXPERIMENTAL 243 00:09:33,400 --> 00:09:35,800 VALIDATION ASSAYS THAT REALLY 244 00:09:35,800 --> 00:09:37,640 DEMONSTRATED THIS PARADIGM IS 245 00:09:37,640 --> 00:09:41,280 ACTUALLY -- HOLDS TOGETHER 246 00:09:41,280 --> 00:09:42,120 PRETTY MUCH EVERY TUMOR TYPE 247 00:09:42,120 --> 00:09:44,080 THAT WE'VE LOOKED AT AT THE 248 00:09:44,080 --> 00:09:46,800 MOLECULAR LEVEL AND SORT OF 249 00:09:46,800 --> 00:09:48,160 MECHANISTIC, AND I THINK I'LL 250 00:09:48,160 --> 00:09:50,000 TRY TO EXPLAIN WHY WE CALL THIS 251 00:09:50,000 --> 00:09:53,640 KIND OF A QUASI MECHANISTIC 252 00:09:53,640 --> 00:09:54,040 MODEL. 253 00:09:54,040 --> 00:09:58,280 THE MORE IMPORTANT THING IS THAT 254 00:09:58,280 --> 00:09:59,400 THESE STUDIES HAVE NOW YIELDED 255 00:09:59,400 --> 00:10:01,400 QUITE A LARGE NUMBER OF CLINICAL 256 00:10:01,400 --> 00:10:03,720 TRIALS AND CLINICAL STUDIES, 257 00:10:03,720 --> 00:10:05,200 CLINICAL END POINTS THAT ARE 258 00:10:05,200 --> 00:10:06,760 CURRENTLY RUNNING, SOME OF THEM 259 00:10:06,760 --> 00:10:08,400 ARE COMPLETED AND I'LL SHOW YOU 260 00:10:08,400 --> 00:10:13,000 THE RESULTS, AND OTHERS WILL 261 00:10:13,000 --> 00:10:14,200 START -- THEY'RE NOT PRESENTED 262 00:10:14,200 --> 00:10:17,040 HERE BUT THEY'LL START IN THE 263 00:10:17,040 --> 00:10:20,640 NEXT FEW MONTHS. 264 00:10:20,640 --> 00:10:21,480 SO ONE OF THE NICE THINGS ABOUT 265 00:10:21,480 --> 00:10:25,040 HAVING A MODEL, HAVING THIS 266 00:10:25,040 --> 00:10:26,640 ONCOTECH TOUR IS WE CAN NOW 267 00:10:26,640 --> 00:10:28,400 START DEVELOPING ALGORITHMS 268 00:10:28,400 --> 00:10:29,920 WHERE WE'RE FIRST GOING TO 269 00:10:29,920 --> 00:10:30,600 RECONSTRUCT THIS ARCHITECTURE 270 00:10:30,600 --> 00:10:31,560 AND WE KNOW IT'S DIFFERENT IN A 271 00:10:31,560 --> 00:10:35,000 WAY THAT IS COMPLETELY 272 00:10:35,000 --> 00:10:36,720 CONTEXT-DEPENDENT SO THEONICS 273 00:10:36,720 --> 00:10:39,200 TEXTURE OF A B CELL IS 274 00:10:39,200 --> 00:10:39,760 COMPLETELY DIFFERENT FROM 275 00:10:39,760 --> 00:10:42,040 THEONICS TECH TOUR OF GLIOMA 276 00:10:42,040 --> 00:10:43,800 CELLS, SO WE USE THIS 277 00:10:43,800 --> 00:10:45,400 METHODOLOGY AND ALL OF THEM HAVE 278 00:10:45,400 --> 00:10:47,320 BEEN EXTENSIVELY VALIDATED 279 00:10:47,320 --> 00:10:48,800 EXPERIMENTALLY, ALL OF THEM HAVE 280 00:10:48,800 --> 00:10:53,480 A 70 TO 80 PERCIVAL DAITION RATE 281 00:10:53,480 --> 00:10:58,200 IN THE EXPERIMENTAL LAB AND THE 282 00:10:58,200 --> 00:10:59,680 FIRST ONE IS AN ALGORITHM THAT 283 00:10:59,680 --> 00:11:00,800 ALLOWS YOU TO TURN A LARGE 284 00:11:00,800 --> 00:11:02,200 NUMBER OF GENE EXPRESSION 285 00:11:02,200 --> 00:11:04,400 PROFILES INTO A FAIRLY ACCURATE 286 00:11:04,400 --> 00:11:06,640 MAP, ABOUT 70 PERCIVAL DAITION 287 00:11:06,640 --> 00:11:10,720 RATE OF THE UNDERLYING 288 00:11:10,720 --> 00:11:11,440 TRANSCRIPTIONAL REGULATION 289 00:11:11,440 --> 00:11:15,800 MECHANICS. 290 00:11:15,800 --> 00:11:18,880 THEN THIS VIPER USES 291 00:11:18,880 --> 00:11:20,120 INTERACTOMES TO MEASURE QUITE 292 00:11:20,120 --> 00:11:21,520 PRECISELY THE TRANSCRIPTIONAL 293 00:11:21,520 --> 00:11:28,800 ACTIVITY OF PROTEINS. 294 00:11:28,800 --> 00:11:31,200 IN THE UPPER PART OF THAT HOUR 295 00:11:31,200 --> 00:11:36,000 GLASS BOTTLE TO IDENTIFY QUITE 296 00:11:36,000 --> 00:11:36,920 EFFECTIVELY ALL THE PROTEINS 297 00:11:36,920 --> 00:11:38,800 THAT HAVE A POTENTIAL MODULATORY 298 00:11:38,800 --> 00:11:40,640 ROLE WHERE BASICALLY THEIR 299 00:11:40,640 --> 00:11:42,240 ACTIVITY OR ABUNDANCE CAN 300 00:11:42,240 --> 00:11:44,080 MODULATE THE ABILITY OF ONE OF 301 00:11:44,080 --> 00:11:47,000 THESE MASTER REGULATORS TO 302 00:11:47,000 --> 00:11:50,200 REGULATE ITS TARGETS AND THEN 303 00:11:50,200 --> 00:11:52,000 FINALLY, FRAPPE WAS THE FIRST IN 304 00:11:52,000 --> 00:11:53,520 DEVELOPING IN COLLABORATION WITH 305 00:11:53,520 --> 00:11:57,360 BARRY TO ACTUALLY PREDICT 306 00:11:57,360 --> 00:11:58,160 PROTEIN-PROTEIN INTERACTION 307 00:11:58,160 --> 00:11:59,360 USING STRUCTURAL BIOLOGY DATA. 308 00:11:59,360 --> 00:12:01,440 LET ME SPEND A COUPLE MINUTES 309 00:12:01,440 --> 00:12:03,120 ON -- BECAUSE IT'S THE WORKFORCE 310 00:12:03,120 --> 00:12:04,320 OF THE LAB AND A LOT OF THE 311 00:12:04,320 --> 00:12:05,760 THINGS I'LL WILL SHOW YOU ARE 312 00:12:05,760 --> 00:12:07,440 BASED ON IT, SO IF I DON'T 313 00:12:07,440 --> 00:12:09,080 EXPLAIN THESE IN A WAY THAT IS 314 00:12:09,080 --> 00:12:09,760 UNDERSTOOD, YOU MAY MISS THE 315 00:12:09,760 --> 00:12:13,920 REST OF THE TALK. 316 00:12:13,920 --> 00:12:17,880 THE WAY IT WORKS IS AKIN TO A 317 00:12:17,880 --> 00:12:23,520 HIGHLY MULTIPLEX GENE REPORTED 318 00:12:23,520 --> 00:12:23,760 ASSAY. 319 00:12:23,760 --> 00:12:27,600 IF YOU USE A -- PROMOTER AND 320 00:12:27,600 --> 00:12:30,640 THEN MEASURE LUMINESCENCE, IF 321 00:12:30,640 --> 00:12:32,360 YOU HAVE LUMINESCENCE AND 322 00:12:32,360 --> 00:12:35,840 RESPONSE TO MAKE -- IF YOU SEE 323 00:12:35,840 --> 00:12:37,920 NONE -- THAT WORKS WELL IF MIC 324 00:12:37,920 --> 00:12:39,880 IS THE ONLY THING THAT MOVES IN 325 00:12:39,880 --> 00:12:42,440 THE CELL, OBVIOUSLY IN RESPONSE 326 00:12:42,440 --> 00:12:44,000 TO OTHER TRANSCRIPTIONAL 327 00:12:44,000 --> 00:12:45,280 REGULATORS, IT WILL NOT WORK 328 00:12:45,280 --> 00:12:47,120 WELL IF EVERYTHING MOVES. 329 00:12:47,120 --> 00:12:49,400 SO WHAT WE TRIED TO DO IS TO 330 00:12:49,400 --> 00:12:51,040 GENERALIZE THIS CONCEPT OF A 331 00:12:51,040 --> 00:12:52,000 GENE REPORTER ASSAY SO THAT WE 332 00:12:52,000 --> 00:12:56,040 NOW CAN USE ABOUT 100 TARGETS OF 333 00:12:56,040 --> 00:12:58,000 EVERY REGULATORY PROTEINS BOTH 334 00:12:58,000 --> 00:13:00,200 POSITIVELY REGULATED AND 335 00:13:00,200 --> 00:13:02,000 REPRESSED AS THE REPORTER ASSAY. 336 00:13:02,000 --> 00:13:03,920 SO FROM THE EXPRESSION OF THESE 337 00:13:03,920 --> 00:13:05,400 TARGETS, ACTUALLY FROM THE 338 00:13:05,400 --> 00:13:06,280 DIFFERENTIAL EXPRESSION OF THESE 339 00:13:06,280 --> 00:13:07,240 TARGETS IN TWO DIFFERENT STATES 340 00:13:07,240 --> 00:13:09,200 OF THE CELL, WE CAN MEASURE THE 341 00:13:09,200 --> 00:13:11,000 DIFFERENTIAL ACTIVITY OF ANY 342 00:13:11,000 --> 00:13:14,440 REGULATORY PROTEINS IN THOSE TWO 343 00:13:14,440 --> 00:13:14,720 COMPLEX. 344 00:13:14,720 --> 00:13:20,440 SO YOU COULD USE TWO ISOGENIC 345 00:13:20,440 --> 00:13:24,440 CONTEXT, ONE WITH -- DRUG 346 00:13:24,440 --> 00:13:27,400 SENSITIVE VERSUS DRUG RESISTANT 347 00:13:27,400 --> 00:13:28,600 AND WE'VE DONE ALL THE STUDIES 348 00:13:28,600 --> 00:13:30,400 AND PUBLISHED THEM AND IN EVERY 349 00:13:30,400 --> 00:13:32,400 SINGLE CASE, WE'VE BEEN ABLE TO 350 00:13:32,400 --> 00:13:33,520 IDENTIFY THE MASTER REGULATORS 351 00:13:33,520 --> 00:13:34,520 AND VALIDATE THEM 352 00:13:34,520 --> 00:13:35,080 EXPERIMENTALLY. 353 00:13:35,080 --> 00:13:37,240 BUT ALSO AS YOU'LL SEE, EVEN IN 354 00:13:37,240 --> 00:13:38,920 VERY EARLY PAPERS, WE SHOW THAT 355 00:13:38,920 --> 00:13:41,560 WE CAN ACTUALLY REPROGRAM THE 356 00:13:41,560 --> 00:13:42,840 STATE OF THE CELL BY TARGETING 357 00:13:42,840 --> 00:13:44,280 THE MASTER REGULATORS BECAUSE 358 00:13:44,280 --> 00:13:46,200 THESE ARE ESSENTIALLY THE 359 00:13:46,200 --> 00:13:49,240 PROTEINS THAT MECHANISTICALLY, I 360 00:13:49,240 --> 00:13:51,400 SHOULD SAY QUASI MECHANISTICALLY 361 00:13:51,400 --> 00:13:52,600 BECAUSE ONLY 70% OF THEIR 362 00:13:52,600 --> 00:13:56,400 TARGETS ARE CORRECT, BUT 363 00:13:56,400 --> 00:13:57,000 MECHANISTICALLY REGULATE THE 364 00:13:57,000 --> 00:13:59,000 STATE OF THE CELL. 365 00:13:59,000 --> 00:14:02,560 SO THERE ARE THREE MAIN HIGH 366 00:14:02,560 --> 00:14:05,600 POTASSIUM THESE THAT 367 00:14:05,600 --> 00:14:07,840 HYPOTHESES THAT STEM FROM THIS 368 00:14:07,840 --> 00:14:08,480 ARCHITECTURE, MAJOR HYPOTHESES 369 00:14:08,480 --> 00:14:10,200 THAT WE HAVE POSTULATED. 370 00:14:10,200 --> 00:14:12,160 THE FIRST ONE IS THAT MASTER 371 00:14:12,160 --> 00:14:16,840 REGULATORS FORM THESE TUMOR 372 00:14:16,840 --> 00:14:18,480 CHECKPOINTS, IN CANCER, WE CALL 373 00:14:18,480 --> 00:14:19,600 THEM REGULATORY CHECKPOINTS WHEN 374 00:14:19,600 --> 00:14:25,000 IT'S NOT IN CANCER, RESPONSIBLE 375 00:14:25,000 --> 00:14:26,600 FOR -- THEY CAN ESSENTIALLY 376 00:14:26,600 --> 00:14:28,360 ADAPT TO ANY PERTURBATION AND 377 00:14:28,360 --> 00:14:29,600 MAINTAIN THE STATE LARGELY 378 00:14:29,600 --> 00:14:30,640 INDEPENDENT OF THESE 379 00:14:30,640 --> 00:14:31,000 PERTURBATIONS. 380 00:14:31,000 --> 00:14:33,120 THESE ARE PAPERS WHERE THAT HAS 381 00:14:33,120 --> 00:14:34,800 BEEN SHOWN, THERE'S MORE -- I'M 382 00:14:34,800 --> 00:14:38,560 JUST SHOWING SOME EXAMPLES. 383 00:14:38,560 --> 00:14:39,280 THE SECOND ONE IS THAT THE 384 00:14:39,280 --> 00:14:41,840 MASTER REGULATORS IN THESE TUMOR 385 00:14:41,840 --> 00:14:46,320 CHECKPOINT MODULES CANNIZE THE 386 00:14:46,320 --> 00:14:48,000 EFFECT OF MOST FUNCTIONAL 387 00:14:48,000 --> 00:14:49,280 MUTATIONAL EVENTS. 388 00:14:49,280 --> 00:14:53,080 THESE PAPERS, PROBABLY BRINGS UP 389 00:14:53,080 --> 00:14:54,760 MEMORIES TO LOU. 390 00:14:54,760 --> 00:14:56,720 THIRD IS MASTER REGULATORS IN 391 00:14:56,720 --> 00:14:58,400 TUMOR CHECKPOINT MODULE 392 00:14:58,400 --> 00:15:00,880 REPRESENT A NEW CLASS OF 393 00:15:00,880 --> 00:15:04,400 PHARMACOLOGICALLY ACTIONABLE 394 00:15:04,400 --> 00:15:05,320 NON-ONCOGENE DEPENDENCIES. 395 00:15:05,320 --> 00:15:07,880 THE PROTEINS THAT ARE -- AND 396 00:15:07,880 --> 00:15:09,040 ALMOST NEVER HARBOR MUTATION 397 00:15:09,040 --> 00:15:11,960 THAT TURN THEM INTO 398 00:15:11,960 --> 00:15:13,320 ESSENTIALLY -- ACTIVE PROTEINS 399 00:15:13,320 --> 00:15:14,080 BECAUSE OF THE MUTATION. 400 00:15:14,080 --> 00:15:16,800 SO LET ME SHOW YOU A CASE STUDY 401 00:15:16,800 --> 00:15:21,560 IN PANCREATIC CANCER. 402 00:15:21,560 --> 00:15:26,320 THIS HAS BEEN LED BY PASS QAL 403 00:15:26,320 --> 00:15:27,360 LAISE IN MY LAB. 404 00:15:27,360 --> 00:15:28,800 TWO OF THE HYPOTHESES ARE 405 00:15:28,800 --> 00:15:31,000 ESSENTIALLY DISCUSSED IN MY DATA 406 00:15:31,000 --> 00:15:32,000 HERE. 407 00:15:32,000 --> 00:15:32,800 PANCREATIC CANCER HAS BEEN 408 00:15:32,800 --> 00:15:36,400 CHALLENGING TO CLASSIFY IN 409 00:15:36,400 --> 00:15:37,800 STRUCTURE BECAUSE IF YOU LOOK AT 410 00:15:37,800 --> 00:15:38,640 THREE DIFFERENT STUDIES DONE 411 00:15:38,640 --> 00:15:42,080 FROM GENE EXPRESSION PROFILES, 412 00:15:42,080 --> 00:15:44,000 THE CLASSES THAT ARE GENERAL 413 00:15:44,000 --> 00:15:45,200 LATED HAVE NO OVERLAP BETWEEN 414 00:15:45,200 --> 00:15:46,080 THE THREE OF THEM, SO YOU CAN 415 00:15:46,080 --> 00:15:49,520 SEE HERE THAT ACCORDING TO FOUR 416 00:15:49,520 --> 00:15:51,400 DIFFERENT STUDIES IN TUMOR AND 417 00:15:51,400 --> 00:15:53,000 STROMA -- OR IN THE TUMOR CELLS, 418 00:15:53,000 --> 00:15:54,360 THERE WERE ZERO OVERLAPPING 419 00:15:54,360 --> 00:15:55,920 GENES THAT WILL DETERMINE THE 420 00:15:55,920 --> 00:15:57,840 IDENTITY OF THE SUBTYPES 421 00:15:57,840 --> 00:15:58,800 IDENTIFIED BY EACH STUDY. 422 00:15:58,800 --> 00:16:00,440 AND WE THOUGHT THAT MAYBE THIS 423 00:16:00,440 --> 00:16:02,560 WAS BECAUSE, ONE, THERE IS 424 00:16:02,560 --> 00:16:05,040 TREMENDOUS STROMAL INFILTRATION, 425 00:16:05,040 --> 00:16:06,040 IN FACT, SOMETIMES THE TUMOR 426 00:16:06,040 --> 00:16:07,840 CELLS ARE ACTUALLY A MINORITY, 427 00:16:07,840 --> 00:16:10,440 OR EVEN A SMALL MINORITY OF ALL 428 00:16:10,440 --> 00:16:12,440 THE CELLS IN THE TUMOR, AND 429 00:16:12,440 --> 00:16:15,040 NUMBER TWO, BECAUSE WITHIN THE 430 00:16:15,040 --> 00:16:16,760 TUMOR ITSELF, THERE MAY BE 431 00:16:16,760 --> 00:16:17,840 MULTIPLE COMPARTMENTS OF CELLS 432 00:16:17,840 --> 00:16:22,680 THAT HAVE STRIKINGLY DISTINCT 433 00:16:22,680 --> 00:16:23,800 MOLECULAR IDENTITIES. 434 00:16:23,800 --> 00:16:26,240 SO TO DO THIS, WE RESORTED TO 435 00:16:26,240 --> 00:16:28,600 STUDY PANCREATIC CANCER AT 436 00:16:28,600 --> 00:16:29,240 SINGLE CELL LEVEL. 437 00:16:29,240 --> 00:16:31,880 I WANT TO SHOW YOU WHAT THE 438 00:16:31,880 --> 00:16:32,880 CHALLENGE IS WHEN YOU GO TO 439 00:16:32,880 --> 00:16:33,280 SINGLE CELLS. 440 00:16:33,280 --> 00:16:37,760 THIS IS DAY TAI DATA PUBLISHED RECENTLY 441 00:16:37,760 --> 00:16:39,240 IN CELL CARCINOMA, IN 442 00:16:39,240 --> 00:16:42,840 COOPERATION WITH CHUCK DRAKE, 443 00:16:42,840 --> 00:16:45,400 PH.D. STUDENTS IN OUR LAB, WHERE 444 00:16:45,400 --> 00:16:47,320 BASICALLY WE TOOK SINGLE CELLS 445 00:16:47,320 --> 00:16:55,560 FROM THESE TUMORS, -- AND THEN 446 00:16:55,560 --> 00:16:57,680 PERFORMED ANALYSES USING EITHER 447 00:16:57,680 --> 00:16:59,800 A CYTEK WHICH USES A PANEL OF 19 448 00:16:59,800 --> 00:17:01,800 MYELOID AND 19 LYMPHOID 449 00:17:01,800 --> 00:17:02,920 ANTIBODY, I'M ONLY SHOWING THE 450 00:17:02,920 --> 00:17:04,160 LYMPHOID HERE, YOU CAN READ THE 451 00:17:04,160 --> 00:17:05,160 PAPER FOR THE OTHER ONES, THEY 452 00:17:05,160 --> 00:17:06,680 BASICALLY TELL THE SAME STORY, 453 00:17:06,680 --> 00:17:08,240 AND THOSE IDENTIFIED FOUR 454 00:17:08,240 --> 00:17:09,000 POPULATIONS THAT YOU WOULD 455 00:17:09,000 --> 00:17:11,120 EXPECT TO FIND IN THESE CELLS. 456 00:17:11,120 --> 00:17:12,480 HOWEVER, IF YOU TAKE THE SAME 457 00:17:12,480 --> 00:17:14,120 CELLS AND SEND THEM TO CHROMIUM 458 00:17:14,120 --> 00:17:15,280 AND YOU LOOK TO THE GENES THAT 459 00:17:15,280 --> 00:17:16,920 ENCODE FOR THESE PROTEINS, YOU 460 00:17:16,920 --> 00:17:18,440 SEE ABSOLUTELY NOTHING. 461 00:17:18,440 --> 00:17:21,200 ESSENTIALLY THERE'S COMPLETE 462 00:17:21,200 --> 00:17:22,600 LOSS OF THE MRNAs THAT ENCODE 463 00:17:22,600 --> 00:17:23,720 FOR THOSE PROTEINS AND, 464 00:17:23,720 --> 00:17:26,200 THEREFORE, YOU CANNOT -- 465 00:17:26,200 --> 00:17:26,640 ANYTHING. 466 00:17:26,640 --> 00:17:28,720 HOWEVER, WHEN YOU NOW RUN VIPER 467 00:17:28,720 --> 00:17:33,200 ON THESE DATA, NOT ONLY YOU CAN 468 00:17:33,200 --> 00:17:37,120 CONSTITUTE BASICALLY THAT WAS 469 00:17:37,120 --> 00:17:42,800 FOUND BY ANTIBODIES, THE 470 00:17:42,800 --> 00:17:45,240 STANDARD VERSUS THE MEAN BECOMES 471 00:17:45,240 --> 00:17:47,400 DRAMATICALLY BETTER WITHIN THE 472 00:17:47,400 --> 00:17:53,600 SUBTYPES, SO CD4 IN THIS CASE, 473 00:17:53,600 --> 00:17:56,640 THE ANTIBODY LEVEL, CD14 FOR 474 00:17:56,640 --> 00:17:57,720 MONOCYTE MACROPHAGES, AGAIN, 475 00:17:57,720 --> 00:18:00,200 VERY NOISY, YOU CAN'T SEE IT IN 476 00:18:00,200 --> 00:18:03,000 THE ANTIBODIES -- FOR THE 477 00:18:03,000 --> 00:18:04,000 CD4 CELLS WHICH IS VERY NOISY 478 00:18:04,000 --> 00:18:05,520 AGAIN. 479 00:18:05,520 --> 00:18:08,960 AND THERE ARE ALSO THINGS LIKE 480 00:18:08,960 --> 00:18:13,880 CD19 WHICH WE DIDN'T DETECT 481 00:18:13,880 --> 00:18:15,400 WELL, WHICH MEANS WE HAVE TO 482 00:18:15,400 --> 00:18:16,320 REFINE -- PROPOSAL WE HAVE FOR 483 00:18:16,320 --> 00:18:18,360 THE NEXT FEW YEARS ADDRESS THAT 484 00:18:18,360 --> 00:18:20,920 QUESTION EXPERIMENTALLY BY 485 00:18:20,920 --> 00:18:23,520 COMBINING -- WITH SINGLE CELL 486 00:18:23,520 --> 00:18:24,080 SILENCING DATA. 487 00:18:24,080 --> 00:18:25,880 SO BASICALLY WE CAN NOW USE 488 00:18:25,880 --> 00:18:28,040 VIPER TO TURN SINGLE CELL DATA 489 00:18:28,040 --> 00:18:30,360 ALMOST LIKE IF IT WERE BULK 490 00:18:30,360 --> 00:18:34,360 TISSUE, SO WE USE DATA FROM A 491 00:18:34,360 --> 00:18:36,120 PRIOR PUBLICATION FROM TUVESON 492 00:18:36,120 --> 00:18:38,920 WHERE WE FOCUS ON THE 493 00:18:38,920 --> 00:18:40,480 FIBROBLASTS, MULTIPLE DIFFERENT 494 00:18:40,480 --> 00:18:45,040 TYPES THAT ARE 495 00:18:45,040 --> 00:18:45,640 IMMUNOSUPPRESSIVE, YOU CAN SEE 496 00:18:45,640 --> 00:18:47,200 HOW SMALL IT IS COMPARED TO ALL 497 00:18:47,200 --> 00:18:52,080 THE CELLS THAT WE IDENTIFIED, WE 498 00:18:52,080 --> 00:18:53,120 VALIDATED THESE WERE ACTUALLY 499 00:18:53,120 --> 00:18:54,800 TUMOR CELLS AND THEN PERFORMED 500 00:18:54,800 --> 00:18:56,000 OUR ANALYSES PERFORMING VIPER 501 00:18:56,000 --> 00:18:57,600 AND CLUSTER, BUT THE CLUSTERING 502 00:18:57,600 --> 00:18:59,720 WAS DONE NOT BY GENE EXPRESSION 503 00:18:59,720 --> 00:19:00,960 PROFILING BUT RATHER BY PROTEIN 504 00:19:00,960 --> 00:19:01,920 ACTIVITY. 505 00:19:01,920 --> 00:19:02,600 WHY IS THAT IMPORTANT? 506 00:19:02,600 --> 00:19:05,080 BECAUSE IF YOU DID THE CLASS -- 507 00:19:05,080 --> 00:19:06,600 EXPRESSION YOU WOULD NOT SEE 508 00:19:06,600 --> 00:19:08,880 THESE THREE POPULATIONS, IN 509 00:19:08,880 --> 00:19:10,440 FACT, FOUR OF THEM, SO PROTEIN 510 00:19:10,440 --> 00:19:13,760 ACTIVITY GENERATED FOUR 511 00:19:13,760 --> 00:19:16,360 SUBTYPES, POPULATION OF SINGLE 512 00:19:16,360 --> 00:19:19,200 CELLS WITH A VERY STRONG SCORE 513 00:19:19,200 --> 00:19:22,200 THAT IS DRAMATICALLY IMPROVING 514 00:19:22,200 --> 00:19:22,640 GENE EXPRESSION. 515 00:19:22,640 --> 00:19:24,400 YOU CAN SEE WE DID ADDITIONAL 516 00:19:24,400 --> 00:19:25,560 ANALYSES, SO THESE ARE THE 517 00:19:25,560 --> 00:19:27,000 DIFFERENT COLORS, BUT YOU CAN 518 00:19:27,000 --> 00:19:29,840 SEE THAT WITHIN THIS CLUSTER OF 519 00:19:29,840 --> 00:19:33,400 GREEN CELLS, THERE WERE ACTUALLY 520 00:19:33,400 --> 00:19:33,840 THREE PEAKS. 521 00:19:33,840 --> 00:19:35,880 SO WE NOW DO THIS PEAK ANALYSIS 522 00:19:35,880 --> 00:19:39,280 BECAUSE ESSENTIALLY FROM THE -- 523 00:19:39,280 --> 00:19:40,320 DISTRIBUTION -- DENSITY OF THE 524 00:19:40,320 --> 00:19:41,720 CELL CORRESPONDS TO A STABLE 525 00:19:41,720 --> 00:19:43,200 STATE, AND THE STABLE STATE ARE 526 00:19:43,200 --> 00:19:45,040 VERY IMPORTANT BECAUSE THE CELLS 527 00:19:45,040 --> 00:19:46,720 ARE NOT UNDER PEAKS. 528 00:19:46,720 --> 00:19:48,480 THEY'RE ESSENTIALLY MOVING VERY 529 00:19:48,480 --> 00:19:51,600 RAPIDLY FROM ONE STABLE -- TO 530 00:19:51,600 --> 00:19:52,680 ANOTHER BUT THEY EVENTUALLY WILL 531 00:19:52,680 --> 00:19:55,960 GET THERE OR THEY HAVE TO 532 00:19:55,960 --> 00:19:58,760 START -- THE ONES IN STABLE 533 00:19:58,760 --> 00:20:00,800 STATE TEND TO PERSIST FOR A LONG 534 00:20:00,800 --> 00:20:01,960 PERIOD OF TIME SO THOSE ARE THE 535 00:20:01,960 --> 00:20:03,520 ONES YOU WANT TO TARGET 536 00:20:03,520 --> 00:20:10,600 PHARMACOLOGICALLY. 537 00:20:10,600 --> 00:20:13,000 DIFFERENTIATE FOR -- STATE AND 538 00:20:13,000 --> 00:20:16,160 THIS IS A VERY INTERESTING -- 539 00:20:16,160 --> 00:20:18,840 PROGENITOR STATE. 540 00:20:18,840 --> 00:20:20,680 WHEN YOU LOOK AT THE -- YOU CAN 541 00:20:20,680 --> 00:20:24,080 SEE YOU HAVE LINEAGE, APS AND 542 00:20:24,080 --> 00:20:24,760 MOS CONVERGING ON THE DIFFERENT 543 00:20:24,760 --> 00:20:26,120 STATE AND THIS IS CONSISTENT 544 00:20:26,120 --> 00:20:32,480 WITH THE VIEW BY -- OF THIS 545 00:20:32,480 --> 00:20:35,840 POPULATION WITH THE APS BEING 546 00:20:35,840 --> 00:20:39,360 THE MOST STEM LIKE AND LIN 547 00:20:39,360 --> 00:20:40,560 AND -- WE LOOK AT THE PRESENCE 548 00:20:40,560 --> 00:20:41,760 OF THESE POPULATION WITHIN 549 00:20:41,760 --> 00:20:42,560 DIFFERENT PATIENTS AND YOU CAN 550 00:20:42,560 --> 00:20:45,000 SEE HERE THAT ESSENTIALLY ALL 551 00:20:45,000 --> 00:20:46,280 THE PATIENT PRESENTED AT LEAST 552 00:20:46,280 --> 00:20:50,400 THREE OF THE FOUR AND IN SOME 553 00:20:50,400 --> 00:20:52,720 CASES ALL FOUR OF THEM, THE APS 554 00:20:52,720 --> 00:20:53,800 ARE MORE RARE, CONSISTENT WITH 555 00:20:53,800 --> 00:20:55,560 THE FACT THAT PROBABLY MORE 556 00:20:55,560 --> 00:20:57,160 STEM-LIKE POPULATIONS OR IN SOME 557 00:20:57,160 --> 00:20:58,200 PATIENTS MAYBE WE HAVEN'T SEEN 558 00:20:58,200 --> 00:21:00,120 THIS BECAUSE WE DIDN'T SEQUENCE 559 00:21:00,120 --> 00:21:00,680 ENOUGH CELLS. 560 00:21:00,680 --> 00:21:02,200 BUT THEN MORE IMPORTANTLY, IF 561 00:21:02,200 --> 00:21:03,400 YOU LOOK AT THE BIOLOGY OF THE 562 00:21:03,400 --> 00:21:04,560 CELLS, YOU CAN SEE THAT THERE 563 00:21:04,560 --> 00:21:07,200 ARE REALLY MARKERS THAT ARE 564 00:21:07,200 --> 00:21:07,600 HIGHLY DISTINCT. 565 00:21:07,600 --> 00:21:08,760 THESE ARE MARKERS THAT YOU CAN 566 00:21:08,760 --> 00:21:12,360 ACTUALLY USE TO -- THE CELLS AND 567 00:21:12,360 --> 00:21:14,400 DIVIDE THE SUBPOPULATION THAT 568 00:21:14,400 --> 00:21:16,400 DISTINGUISH THESE FOUR DIFFERENT 569 00:21:16,400 --> 00:21:18,040 SUBPOPULATIONS, AND JUST AS A 570 00:21:18,040 --> 00:21:20,760 SUMMARY HERE, YOU CAN SEE THAT 571 00:21:20,760 --> 00:21:21,920 THIS POPULATION HERE ARE VERY, 572 00:21:21,920 --> 00:21:23,280 VERY SMALL NUMBER OF CELLS IS 573 00:21:23,280 --> 00:21:26,920 ACTUALLY EXTREMELY DISTINCT AND 574 00:21:26,920 --> 00:21:29,480 THESE ARE -- THREE OF THESE 575 00:21:29,480 --> 00:21:32,000 MARKERS ARE CLASSICAL MARKERS OF 576 00:21:32,000 --> 00:21:36,120 ADM, YOU CAN SEE HERE 577 00:21:36,120 --> 00:21:38,160 REPRESENTED -- AND THESE CELLS 578 00:21:38,160 --> 00:21:40,000 THE ONE CUT WHICH YOU CAN SEE 579 00:21:40,000 --> 00:21:41,400 HERE BASICALLY ACTIVATED ONLY 580 00:21:41,400 --> 00:21:46,400 THE CELLS AND THE SOX 9. 581 00:21:46,400 --> 00:21:48,400 SO THIS ANALYSIS REALLY ALLOWS 582 00:21:48,400 --> 00:21:50,040 YOU TO DISCOVER BIOLOGY THAT 583 00:21:50,040 --> 00:21:51,600 THEN CAN BE VALIDATED THAT WE 584 00:21:51,600 --> 00:21:53,480 MAY HAVE COMPLETELY MISSED IF 585 00:21:53,480 --> 00:21:55,800 YOU WERE TO USE GENE EXPRESSION 586 00:21:55,800 --> 00:21:56,400 PROFILE. 587 00:21:56,400 --> 00:22:01,880 WE THEN WENT TO TWO -- OF SINGLE 588 00:22:01,880 --> 00:22:05,120 CELLS. 589 00:22:05,120 --> 00:22:05,640 CSY. 590 00:22:05,640 --> 00:22:07,240 YOU CAN SEE SINGLE CELLS FROM 591 00:22:07,240 --> 00:22:08,480 THIS DATASET COMPLETELY 592 00:22:08,480 --> 00:22:11,400 VALIDATED THE STRUCTURE IN THE 593 00:22:11,400 --> 00:22:14,080 FOUR CLUSTERS WITH THE 594 00:22:14,080 --> 00:22:16,600 DIFFERENTIATED PROGENITOR CELLS 595 00:22:16,600 --> 00:22:17,440 FORMING THE MAJORITY OF THE 596 00:22:17,440 --> 00:22:21,320 CELLS AND THEN LINEAGE -- 597 00:22:21,320 --> 00:22:22,280 REPRESENTING THE REST. 598 00:22:22,280 --> 00:22:23,680 AND THIS WAS VERY HELPFUL 599 00:22:23,680 --> 00:22:26,240 BECAUSE IN EVERY SINGLE STUDY 600 00:22:26,240 --> 00:22:28,920 WE'VE DONE, WHEN WE DO THESE 601 00:22:28,920 --> 00:22:30,000 ANALYSES AT THE PROTEIN LEVEL AS 602 00:22:30,000 --> 00:22:31,240 HE OH POSE TODAY GENE EXPRESSION 603 00:22:31,240 --> 00:22:33,040 LEVEL WE FIND TREMENDOUS 604 00:22:33,040 --> 00:22:34,440 CONSISTENCY IN DIFFERENT 605 00:22:34,440 --> 00:22:34,720 DATABASES. 606 00:22:34,720 --> 00:22:37,440 SO THESE WERE 35 PATIENTS, 607 00:22:37,440 --> 00:22:38,240 21,000 SINGLE CELLS. 608 00:22:38,240 --> 00:22:42,960 SO THEN WE WENT TO THE BULK 609 00:22:42,960 --> 00:22:45,000 COHORT AND ASK IF WE CAN 610 00:22:45,000 --> 00:22:46,240 RECAPITULATE SOME OF THESE -- 611 00:22:46,240 --> 00:22:48,600 SUBTYPES SO IF YOU ACTUALLY JUST 612 00:22:48,600 --> 00:22:50,800 DO STRATIFICATION BY BULK 613 00:22:50,800 --> 00:22:53,120 PROFILE BASED PROTEIN ACTIVITY, 614 00:22:53,120 --> 00:22:55,320 YOU SEE THAT BOTH ICGC WHICH IS 615 00:22:55,320 --> 00:23:01,040 THE GREEN AND TCGAs 25 616 00:23:01,040 --> 00:23:02,560 ACCORDING TO THESE PROTEINS HERE 617 00:23:02,560 --> 00:23:03,960 AND AGAIN THESE WERE INCREDIBLY 618 00:23:03,960 --> 00:23:05,600 ENRICHED IN THE MASTER 619 00:23:05,600 --> 00:23:09,360 REGULATORS OF THE LINEAGE AND 620 00:23:09,360 --> 00:23:12,600 MORPHOGENIC SUBTYPE. 621 00:23:12,600 --> 00:23:14,000 HOWEVER, THE -- SIMPLY COULD NOT 622 00:23:14,000 --> 00:23:15,240 BE IDENTIFIED IN ANY OF THE 623 00:23:15,240 --> 00:23:15,600 DATASETS. 624 00:23:15,600 --> 00:23:17,320 THIS WAS NOT A DATASET GENERATED 625 00:23:17,320 --> 00:23:20,680 BY US, BY LASER MICRO RESECTION 626 00:23:20,680 --> 00:23:25,760 IN CONNECTION -- 200 PANCREATIC 627 00:23:25,760 --> 00:23:29,400 CANCER PATIENTS, BASICALLY YOU 628 00:23:29,400 --> 00:23:31,400 CAN SEE THE PROTEINS YOU FIND IN 629 00:23:31,400 --> 00:23:34,640 THE DATASETS ARE 630 00:23:34,640 --> 00:23:35,560 DIFFERENTIATING -- ARE 631 00:23:35,560 --> 00:23:36,800 INCREDIBLY OVERLAPPING IN TERMS 632 00:23:36,800 --> 00:23:37,960 OF THEIR MASTER REGULATOR. 633 00:23:37,960 --> 00:23:40,400 SO AGAIN, ALL THE OTHER SINGLE 634 00:23:40,400 --> 00:23:43,960 CELL ACTIVITY MACRO FILE NOW 635 00:23:43,960 --> 00:23:45,160 INSTEAD OF FINDING GENE 636 00:23:45,160 --> 00:23:48,400 EXPRESSION PANELS THAT HAVE NO 637 00:23:48,400 --> 00:23:51,000 OVERLAP, MASTER -- PANELS THAT 638 00:23:51,000 --> 00:23:52,000 ARE VIRTUALLY IDENTICAL ACROSS 639 00:23:52,000 --> 00:23:52,880 THESE TWO DIFFERENT TUMORS. 640 00:23:52,880 --> 00:23:54,600 SO WE WENT TO CELL LINES BECAUSE 641 00:23:54,600 --> 00:23:56,080 YOU NEED CELL LINES TO DO SOME 642 00:23:56,080 --> 00:23:57,120 OF THE EXPERIMENTS SO WE 643 00:23:57,120 --> 00:23:58,560 IDENTIFIED THAT IN A COLLECTION 644 00:23:58,560 --> 00:23:59,880 OF EIGHT DIFFERENT CELL LINES, 645 00:23:59,880 --> 00:24:03,600 WE COULD FIND SINGLE CELL 646 00:24:03,600 --> 00:24:06,960 REPRESENTATIVE LINEAGE, DPS, APS 647 00:24:06,960 --> 00:24:08,160 AND MORPHOGENIC SUBTYPES AND 648 00:24:08,160 --> 00:24:09,560 THIS GIVES YOU THE IDENTITY OF 649 00:24:09,560 --> 00:24:12,320 THE LINES, FOR INSTANCE -- WAS 650 00:24:12,320 --> 00:24:14,200 PREDOMINANTLY LINEAGE AND DPS, 651 00:24:14,200 --> 00:24:16,600 YOU CAN SEE ESSENTIALLY ALONG 652 00:24:16,600 --> 00:24:20,800 THIS AXIS, WHILE KP4, VERY 653 00:24:20,800 --> 00:24:23,120 AGGRESSIVE CELL LINE, WAS MOSTLY 654 00:24:23,120 --> 00:24:26,360 MORPHOGENIC -- STORY, THIS 655 00:24:26,360 --> 00:24:27,880 LINEAGE IN -- THIS IS 656 00:24:27,880 --> 00:24:28,840 MORPHOGENIC IN DPS. 657 00:24:28,840 --> 00:24:32,360 THEN WE DID LINEAGE BARCODE -- 658 00:24:32,360 --> 00:24:36,000 TRACING ASSAYS BECAUSE WE WANTED 659 00:24:36,000 --> 00:24:38,200 TO KNOW WHETHER THE PLOT I 660 00:24:38,200 --> 00:24:40,080 SHOWED YOU BEFORE BASED ON 661 00:24:40,080 --> 00:24:42,040 ANTEROPEE WAS HOLDING -- 662 00:24:42,040 --> 00:24:43,320 DIFFERENTIATION SO WHETHER THE 663 00:24:43,320 --> 00:24:44,480 PLASTICITY WAS GOING TWO WAYS OR 664 00:24:44,480 --> 00:24:45,040 ONLY ONE WAY. 665 00:24:45,040 --> 00:24:49,800 SO THIS IS IN KP4 CELL, ALSO 666 00:24:49,800 --> 00:24:52,720 FOR -- TWO CELLS THAT ARE THEIR 667 00:24:52,720 --> 00:24:54,600 LINEAGE, BASICALLY -- THESE ARE 668 00:24:54,600 --> 00:24:59,400 BARCODES THAT YOU SEE AT TIME 1, 669 00:24:59,400 --> 00:25:00,680 48 HOURS AFTER THE START, AND 670 00:25:00,680 --> 00:25:02,000 ONLY IN THIS POPULATION, AND NOW 671 00:25:02,000 --> 00:25:05,000 YOU CAN SEE THAT TIME .2, WHICH 672 00:25:05,000 --> 00:25:07,960 IS ABOUT 25 DAYS, YOU SEE THAT 673 00:25:07,960 --> 00:25:10,200 THESE BARCODE ARE REPROGRAMMED 674 00:25:10,200 --> 00:25:12,080 TO THE DPS CELLS. 675 00:25:12,080 --> 00:25:14,720 SO OBVIOUSLY WE CAN SEE THERE IS 676 00:25:14,720 --> 00:25:16,640 TREMENDOUS -- BUT WHEN YOU 677 00:25:16,640 --> 00:25:17,640 ACTUALLY LOOK AT THE OPPOSITE, 678 00:25:17,640 --> 00:25:19,400 THESE BARCODES THAT WERE IN THE 679 00:25:19,400 --> 00:25:22,280 DPS CELLS THEN FOUND IN THE 680 00:25:22,280 --> 00:25:23,200 MORPHOGENIC CELLS, YOU ALSO SEE 681 00:25:23,200 --> 00:25:23,600 THAT. 682 00:25:23,600 --> 00:25:25,120 SO THIS SHOWS THESE CELLS ARE 683 00:25:25,120 --> 00:25:32,240 REALLY SPON TAPE SPONTANEOUSLY 684 00:25:32,240 --> 00:25:33,240 REPROGRAMMING -- DIFFERENT 685 00:25:33,240 --> 00:25:33,800 KINETICS. 686 00:25:33,800 --> 00:25:36,400 SO WE THEN MOVE TO SELECTING THE 687 00:25:36,400 --> 00:25:38,480 THREE MOST MORPHOGENIC AND THE 688 00:25:38,480 --> 00:25:42,520 THREE MOST LINEAGE CELLS -- 689 00:25:42,520 --> 00:25:43,840 COLLECTION OF PANCREATIC CANCER 690 00:25:43,840 --> 00:25:45,240 CELLS, YOU CAN SEE THE THREE 691 00:25:45,240 --> 00:25:46,520 THAT ARE MORPHOGENIC AND THE 692 00:25:46,520 --> 00:25:49,160 THREE THAT ARE MORE 693 00:25:49,160 --> 00:25:51,560 LINEAGE-LIKE, AND THEN WE 694 00:25:51,560 --> 00:25:53,600 PERFORMED CRISPR I-BASED 695 00:25:53,600 --> 00:25:55,200 SILENCING WITH THE LIBRARY OF 696 00:25:55,200 --> 00:25:56,840 ALL THE TRANSCRIPTIONAL 697 00:25:56,840 --> 00:25:59,160 REGULATORS AND COTRANSCRIPTION 698 00:25:59,160 --> 00:26:01,080 FACTORS AND NEGATIVE CONTROL, OF 699 00:26:01,080 --> 00:26:02,720 COURSE, AND THEN WE USED THIS 700 00:26:02,720 --> 00:26:04,040 LIBRARY TO LOOK AT TWO THINGS. 701 00:26:04,040 --> 00:26:06,400 THE FIRST ONE WE ASKED, ARE 702 00:26:06,400 --> 00:26:09,600 THERE MASTER REGULATORS THAT -- 703 00:26:09,600 --> 00:26:11,720 THERE IS ACTUALLY 704 00:26:11,720 --> 00:26:12,120 SUBTYPE-SPECIFIC. 705 00:26:12,120 --> 00:26:15,280 SO WE GENERATED TO COMPARE THE 706 00:26:15,280 --> 00:26:17,160 LINEAGE VERSUS MORPHOGENIC CELLS 707 00:26:17,160 --> 00:26:19,600 AND WE ASKED WHETHER THEY WERE 708 00:26:19,600 --> 00:26:21,520 ENRICHED -- KNOCKOUT AND BY 709 00:26:21,520 --> 00:26:23,440 CRISPRI AND YOU CAN SEE STRIKING 710 00:26:23,440 --> 00:26:25,200 ENRICHMENT, 10 TO THE MINUS 6 IN 711 00:26:25,200 --> 00:26:27,000 TERMS OF THE P VALUE OF THE 712 00:26:27,000 --> 00:26:28,400 ENRICHMENT OF THE PREDICTED 713 00:26:28,400 --> 00:26:32,120 MASTER REGULATORS, IN ONE 714 00:26:32,120 --> 00:26:33,160 LINEAGE VERSUS THE OTHER. 715 00:26:33,160 --> 00:26:35,600 THEN WE ACTUALLY ASKED THERE ARE 716 00:26:35,600 --> 00:26:36,840 SOME COMMON DEPENDENCIES THAT 717 00:26:36,840 --> 00:26:38,920 CAN BE FOUND BY, FOR INSTANCE, 718 00:26:38,920 --> 00:26:40,400 COMPARING BOTH MORPHOGENIC CELLS 719 00:26:40,400 --> 00:26:43,360 TO THEIR NORMAL PANCREATIC 720 00:26:43,360 --> 00:26:44,880 TISSUE COUNTERPART IN G TECH, 721 00:26:44,880 --> 00:26:46,800 AND SO THIS IS A SIGNATURE OR 722 00:26:46,800 --> 00:26:49,000 ESSENTIALLY A CANCER PROGRESSION 723 00:26:49,000 --> 00:26:51,800 AND AGAIN THERE WAS REALLY A 724 00:26:51,800 --> 00:26:52,760 STRIKING ENRICHMENT OF THE TOP 725 00:26:52,760 --> 00:26:54,680 MASTER REGULATORS IN GENES THAT 726 00:26:54,680 --> 00:26:58,000 ARE ESSENTIAL IN ALL THE SEQ 727 00:26:58,000 --> 00:26:59,680 CELL LINES AS OPPOSED TO JUST 728 00:26:59,680 --> 00:27:01,320 ONE SUBTYPE VERSUS THE OTHER. 729 00:27:01,320 --> 00:27:05,040 SO THIS BASICALLY SUPPORTS THE 730 00:27:05,040 --> 00:27:08,400 HYPOTHESIS OF THE NEW CLASS OF 731 00:27:08,400 --> 00:27:09,240 NON-ONCOGENE DEPENDENCIES. 732 00:27:09,240 --> 00:27:10,680 THEY'RE VERY RICH, I'LL GIVE YOU 733 00:27:10,680 --> 00:27:12,280 A PAPER ON THAT, WHERE THIS HAS 734 00:27:12,280 --> 00:27:13,080 BEEN SHOWN. 735 00:27:13,080 --> 00:27:14,440 THE MORE INTERESTING THING IS TO 736 00:27:14,440 --> 00:27:16,920 ACTUALLY SHOW THAT THESE GENES, 737 00:27:16,920 --> 00:27:19,080 ACTUALLY THESE PROTEINS 738 00:27:19,080 --> 00:27:19,760 MECHANISTICALLY CONTROL THE 739 00:27:19,760 --> 00:27:20,880 STATE OF THE CELL. 740 00:27:20,880 --> 00:27:23,200 TO DO THAT, WE ECTOPICALLY 741 00:27:23,200 --> 00:27:24,720 EXPRESS THE MASTER REGULATORS OF 742 00:27:24,720 --> 00:27:27,880 THE LINEAGE SUBTYPE IN KP4 CELLS 743 00:27:27,880 --> 00:27:30,040 WHICH ARE STRONGLY MORPHOGENIC. 744 00:27:30,040 --> 00:27:32,160 SO THIS IS THE SIGNATURE, THIS 745 00:27:32,160 --> 00:27:33,760 IS THE ACTIVITY OF THE PROTEINS 746 00:27:33,760 --> 00:27:38,760 IN THE KP4 CELLS, ON THE 747 00:27:38,760 --> 00:27:39,920 SIGNATURE -- MORPHOGENIC VERSUS 748 00:27:39,920 --> 00:27:41,520 LINEAGE, AND THIS IS THE 749 00:27:41,520 --> 00:27:42,640 SIGNATURE OF THE LINEAGE 750 00:27:42,640 --> 00:27:42,920 SUBTYPES. 751 00:27:42,920 --> 00:27:48,400 SO YOU CAN SEE HERE THAT -- 752 00:27:48,400 --> 00:27:51,040 VIRTUALLY EVERY MASTER 753 00:27:51,040 --> 00:27:51,520 REGULATORS SIGNIFICANT 754 00:27:51,520 --> 00:27:52,400 ACTIVATION OF THE PROTEINS THAT 755 00:27:52,400 --> 00:27:57,800 WERE INHIBITED, DOWNACTIVATED IN 756 00:27:57,800 --> 00:27:58,600 THE MORPHOGENIC SUBTYPES BUT 757 00:27:58,600 --> 00:28:00,400 THEY HAD A MIXED RESULT OF THE 758 00:28:00,400 --> 00:28:02,840 PROTEINS THAT WERE ACTIVATED, 759 00:28:02,840 --> 00:28:06,840 BUT ONE OF THEM ACTUALLY INDUCE 760 00:28:06,840 --> 00:28:09,960 COMPLETE REPROGRAMMING SO THESE 761 00:28:09,960 --> 00:28:12,480 CELLS AFTER INDUCTION LOOKED 762 00:28:12,480 --> 00:28:15,720 EXACTLY LIKE LINEAGE CELL, 763 00:28:15,720 --> 00:28:16,120 MORPHOGENIC CELL. 764 00:28:16,120 --> 00:28:18,200 WE ACTUALLY HAD THE RNA SEQ 765 00:28:18,200 --> 00:28:21,920 AFTER EACH ONE OF THESE ECTOPIC 766 00:28:21,920 --> 00:28:23,200 EXPRESSIONS, YOU CAN ACTUALLY 767 00:28:23,200 --> 00:28:24,440 RECONSTRUCT THE FULL 768 00:28:24,440 --> 00:28:26,680 ARCHITECTURE OF THE MASTER REG 769 00:28:26,680 --> 00:28:28,400 LATER MODULES AND YOU NOW 770 00:28:28,400 --> 00:28:30,840 UNDERSTAND MECHANISTICALLY 771 00:28:30,840 --> 00:28:31,800 WHY -- IS RESPONSIBLE FOR 772 00:28:31,800 --> 00:28:35,520 DRIVING THAT TRANSITION 773 00:28:35,520 --> 00:28:36,400 BECAUSE -- ACTUALLY REPRESSES 774 00:28:36,400 --> 00:28:38,200 ALL THE MASTER REGULATORS OR A 775 00:28:38,200 --> 00:28:39,640 LARGE NUMBER OF THE MASTER 776 00:28:39,640 --> 00:28:41,440 REGULATORS OF THE MORPHOGENIC 777 00:28:41,440 --> 00:28:42,680 LINEAGE AND ACTIVATE A LARGE 778 00:28:42,680 --> 00:28:46,800 NUMBER OF MASTER REGULATORS OF 779 00:28:46,800 --> 00:28:50,200 THE LINEAGE, AND THE ONES THAT 780 00:28:50,200 --> 00:28:52,160 IT ALSO REGULATES INDIRECTLY 781 00:28:52,160 --> 00:28:52,760 REGULATES THE ONES THAT ARE 782 00:28:52,760 --> 00:28:53,200 MISSING. 783 00:28:53,200 --> 00:28:56,240 SO TAKEN TOGETHER, THIS INDUCES 784 00:28:56,240 --> 00:28:57,680 REALLY DRAMATIC REPROGRAMMING. 785 00:28:57,680 --> 00:29:01,680 WE THEN ASK, IS THIS ACTUALLY 786 00:29:01,680 --> 00:29:05,680 DUE TO THE -- WE DIDN'T SEE THE 787 00:29:05,680 --> 00:29:06,600 REPROGRAMMING BECAUSE THE EFFECT 788 00:29:06,600 --> 00:29:08,560 OCCURRED ONLY IN A SUBSET OF THE 789 00:29:08,560 --> 00:29:10,240 CELLS, SOMETIMES WHEN YOU 790 00:29:10,240 --> 00:29:11,640 REPROGRAM YOU HAVE LOW 791 00:29:11,640 --> 00:29:12,960 EFFICIENCY SO WE REPEATED THESE 792 00:29:12,960 --> 00:29:15,600 AT THE SINGLE CELL LEVEL AND WE 793 00:29:15,600 --> 00:29:18,000 OVEREXPRESSED GENES WITH AN MY 794 00:29:18,000 --> 00:29:19,600 OF 1 SO THAT SOME CELLS RECEIVED 795 00:29:19,600 --> 00:29:23,360 MORE THAN ONE CDNA, SO YOU CAN 796 00:29:23,360 --> 00:29:25,280 SEE HERE THAT ESSENTIALLY A LOT 797 00:29:25,280 --> 00:29:26,400 OF DIFFERENT COMBINATIONS AND 798 00:29:26,400 --> 00:29:28,240 SINGLE GENES ACTUALLY 799 00:29:28,240 --> 00:29:29,000 STATISTICALLY SIGNIFICANTLY 800 00:29:29,000 --> 00:29:33,560 REPROGRAM THE CELL BUT WITH 801 00:29:33,560 --> 00:29:34,720 DIFFERENT EFFICACY AND 802 00:29:34,720 --> 00:29:37,800 EFFICIENCY. 803 00:29:37,800 --> 00:29:43,320 SO OL2 IS HERE, F1A DRAMATICALLY 804 00:29:43,320 --> 00:29:45,400 INCREASES THE EFFICACY OF THE 805 00:29:45,400 --> 00:29:46,960 REPROGRAMMING, AND THIS IS JUST 806 00:29:46,960 --> 00:29:50,600 A SUBSET OF THE CELLS THAT WE 807 00:29:50,600 --> 00:29:51,960 CALL REPROGRAMMED TO THE LINEAR 808 00:29:51,960 --> 00:29:52,280 SUBTYPE. 809 00:29:52,280 --> 00:29:55,400 SO THIS, I HOPE, DEMONSTRATES 810 00:29:55,400 --> 00:29:56,960 THAT THESE PROTEINS ARE 811 00:29:56,960 --> 00:29:59,160 RESPONSIBLE REALLY FOR HOMO 812 00:29:59,160 --> 00:30:00,160 STATICALLY MAINTAINING THE STATE 813 00:30:00,160 --> 00:30:01,800 OF THE CELLS AND BY GENETICALLY 814 00:30:01,800 --> 00:30:04,000 INTERFERING WITH THEM, YOU CAN 815 00:30:04,000 --> 00:30:07,840 REPROGRAM A CELL AS YOU WANT. 816 00:30:07,840 --> 00:30:10,400 SO THIS WAS PUBLISHED IN CELL 817 00:30:10,400 --> 00:30:14,280 ABOUT EIGHT MONTHS AGO, WHERE WE 818 00:30:14,280 --> 00:30:16,400 ESSENTIALLY ASKED AND 819 00:30:16,400 --> 00:30:17,720 GENERALIZED THESE TWO TUMORS, 820 00:30:17,720 --> 00:30:22,840 CAN WE USE THE ONCO -- IGNORING 821 00:30:22,840 --> 00:30:24,360 ANYTHING WE KNOW ABOUT SUBTYPES, 822 00:30:24,360 --> 00:30:27,000 ANYTHING WE KNOW ABOUT TUMOR 823 00:30:27,000 --> 00:30:27,920 TYPE, ET CETERA, OTHER THAN IN 824 00:30:27,920 --> 00:30:30,400 USING OBVIOUSLY THE RIGHT 825 00:30:30,400 --> 00:30:32,920 REGULATORY NETWORKS FROM A 826 00:30:32,920 --> 00:30:36,200 TISSUE THAT IS MAPPED. 827 00:30:36,200 --> 00:30:38,800 -- NOW AN INDEPENDENT 828 00:30:38,800 --> 00:30:41,000 INVESTIGATOR AT BARCELONA, 829 00:30:41,000 --> 00:30:44,800 AND -- FROM MY LAB, MARIANO IS 830 00:30:44,800 --> 00:30:49,160 THE CHIEF SCIENTIFIC OFFICER, SO 831 00:30:49,160 --> 00:30:53,680 WHAT YOU SEE HERE IS THAT 832 00:30:53,680 --> 00:30:55,920 INSTEAD OF WHAT DOING WHAT WE 833 00:30:55,920 --> 00:30:58,160 DID IN A CELL PAPER, GOING FROM 834 00:30:58,160 --> 00:30:59,200 GENE EXPRESSION TO THE MASTER 835 00:30:59,200 --> 00:31:03,400 REG A LITTLE TORE USING 836 00:31:03,400 --> 00:31:04,640 REGULATOR -- BASICALLY IDENTIFY 837 00:31:04,640 --> 00:31:07,000 ALL THE POSSIBLE MUTATION THAT 838 00:31:07,000 --> 00:31:08,400 CAN AFFECT THE ACTIVITY OF THE 839 00:31:08,400 --> 00:31:10,640 MASTER REGULATORS, WHAT WE DID 840 00:31:10,640 --> 00:31:12,200 HERE, WE PUT THE MASTER 841 00:31:12,200 --> 00:31:15,400 REGULATORS IN THE CROSSHATCH OF 842 00:31:15,400 --> 00:31:16,480 BOTH -- GENETICS IN THE 843 00:31:16,480 --> 00:31:17,160 DOWNSTREAM TRANSCRIPTIONAL 844 00:31:17,160 --> 00:31:17,440 REGULATION. 845 00:31:17,440 --> 00:31:19,400 THIS ALLOWED US TO IDENTIFY, TO 846 00:31:19,400 --> 00:31:21,680 STRATIFY THE ENTIRETY OF TCJ1 847 00:31:21,680 --> 00:31:24,400 SAMPLE AT A TIME, IN 112 TUMOR 848 00:31:24,400 --> 00:31:26,120 SUBTYPES, AND THE FIRST TRACKING 849 00:31:26,120 --> 00:31:27,600 THINGS WAS THAT WHEN YOU LOOK AT 850 00:31:27,600 --> 00:31:30,240 EACH ONE OF THE TUMOR TYPE, ALL 851 00:31:30,240 --> 00:31:34,400 OF THEM EXCEPT -- IDENTIFY 852 00:31:34,400 --> 00:31:36,240 SUBTYPES THAT HAD STATISTICALLY 853 00:31:36,240 --> 00:31:37,640 SIGNIFICANT DIFFERENTIAL 854 00:31:37,640 --> 00:31:38,760 OUTCOME. 855 00:31:38,760 --> 00:31:42,480 THIS HAS BEEN VERY 856 00:31:42,480 --> 00:31:44,960 CHALLENGING -- IN PROSTATE 857 00:31:44,960 --> 00:31:45,960 CANCER I WOULD CHALLENGE YOU TO 858 00:31:45,960 --> 00:31:49,640 FIND ANY -- THAT IDENTIFIES -- J 859 00:31:49,640 --> 00:31:50,840 SUBTYPES THAT HAVE DIFFERENT 860 00:31:50,840 --> 00:31:51,080 OUTCOME. 861 00:31:51,080 --> 00:31:52,520 THIS IS AN EXAMPLE FOR KIDNEY 862 00:31:52,520 --> 00:31:54,000 CANCER, WHERE WE FOUND FIVE 863 00:31:54,000 --> 00:31:56,400 SUBTYPES AND YOU LOOKED AT 864 00:31:56,400 --> 00:31:58,400 S3 VERSUS 5, REALLY STRIKING 865 00:31:58,400 --> 00:32:00,840 DITCH RENGS 866 00:32:00,840 --> 00:32:02,400 DIFFERENTIAL OUTCOME. 867 00:32:02,400 --> 00:32:04,080 THE OTHER -- BY PROTEIN 868 00:32:04,080 --> 00:32:07,880 ACTIVITY, THESE ARE THE BLUE 869 00:32:07,880 --> 00:32:10,040 CLUSTERS, THE BLUE -- PLOT, THIS 870 00:32:10,040 --> 00:32:11,480 IS A SILHOUETTE SCORE THAT TELLS 871 00:32:11,480 --> 00:32:12,840 YOU HOW TIGHT THESE CLUSTERS 872 00:32:12,840 --> 00:32:13,040 ARE. 873 00:32:13,040 --> 00:32:16,400 THE RED LINE, THE DOTTED LINE IS 874 00:32:16,400 --> 00:32:17,800 BASICALLY THE LINE FOR 875 00:32:17,800 --> 00:32:19,800 STATISTICAL SIGNIFICANCE. 876 00:32:19,800 --> 00:32:22,320 ANYTHING ABOVE IS SIGNIFICANT, 877 00:32:22,320 --> 00:32:25,200 THE RED PLOT, WHAT YOU GET 878 00:32:25,200 --> 00:32:26,120 FROM -- EXPRESSION ANALYSIS. 879 00:32:26,120 --> 00:32:28,800 SO THE CLUSTERING OF PCJ BY GENE 880 00:32:28,800 --> 00:32:30,120 EXPRESSION REALLY DOES NOT 881 00:32:30,120 --> 00:32:31,320 PRODUCE CLUSTERS THAT ARE VERY 882 00:32:31,320 --> 00:32:32,360 TIGHT. 883 00:32:32,360 --> 00:32:33,960 SO THIS IS KIND OF THE ONE SLIDE 884 00:32:33,960 --> 00:32:35,800 IN TERMS OF SHOWING THAT THE 885 00:32:35,800 --> 00:32:36,760 GENETIC IS UPSTREAM OF THE 886 00:32:36,760 --> 00:32:37,960 MASTER REGULATORS, BECAUSE 887 00:32:37,960 --> 00:32:39,280 BASICALLY WE ASKED THE QUESTION 888 00:32:39,280 --> 00:32:41,400 AND WE ASK THIS QUESTION ONE 889 00:32:41,400 --> 00:32:44,960 SAMPLE AT A TIME, HOW MANY 890 00:32:44,960 --> 00:32:46,120 GENETIC ALTERATION THAT WE CAN 891 00:32:46,120 --> 00:32:53,280 IDENTIFY IN THAT SAMPLE BY GI 892 00:32:53,280 --> 00:32:56,160 STIC 2 AND CHASM CAN BE PLACED 893 00:32:56,160 --> 00:32:58,040 UPSTREAM OF THE SAMPLE. 894 00:32:58,040 --> 00:33:01,120 EACH COLOR LINE IS ONE OF THE 895 00:33:01,120 --> 00:33:02,200 112 SUBTYPES. 896 00:33:02,200 --> 00:33:03,880 OVARIAN CANCER WHERE THERE'S AN 897 00:33:03,880 --> 00:33:04,840 ENORMOUS NUMBER OF PASSENGER 898 00:33:04,840 --> 00:33:06,800 MUTATION AND STRUCTURAL EVENTS, 899 00:33:06,800 --> 00:33:09,920 THE SATURATION WAS DOUBLY FAST, 900 00:33:09,920 --> 00:33:12,320 BUT YOU GET TO EXPLAIN ABOUT 901 00:33:12,320 --> 00:33:14,600 100% OF THE MUTATION WITH 902 00:33:14,600 --> 00:33:16,520 SOMEWHERE AROUND 15 TO 25 MASTER 903 00:33:16,520 --> 00:33:17,440 REGULATORS. 904 00:33:17,440 --> 00:33:19,200 SO THIS BASICALLY IS A 905 00:33:19,200 --> 00:33:20,200 SATURATION CURVE, TELLS YOU HOW 906 00:33:20,200 --> 00:33:22,360 MUCH OF THE GENETICS IS UPSTREAM 907 00:33:22,360 --> 00:33:24,000 OF HOW MANY OF THE MASTER 908 00:33:24,000 --> 00:33:24,600 REGULATORS. 909 00:33:24,600 --> 00:33:25,600 THIS SIDE IS VERY IMPORTANT 910 00:33:25,600 --> 00:33:26,560 BECAUSE IT TELLS US THAT WHEN WE 911 00:33:26,560 --> 00:33:28,280 WANT TO REALLY KILL A TUMOR AND 912 00:33:28,280 --> 00:33:31,200 WE WANT TO DO THIS BY TARGETING 913 00:33:31,200 --> 00:33:32,560 THE MASTER REGULATORS THAT ARE 914 00:33:32,560 --> 00:33:34,280 BARELY ACTIVATED AS A RESULT OF 915 00:33:34,280 --> 00:33:36,280 THE MUTATIONS IN THAT SAMPLES, 916 00:33:36,280 --> 00:33:38,280 WE REALLY HAVE TO FOCUS ON 917 00:33:38,280 --> 00:33:40,600 AVERAGE OF THE TOP 25 MOST 918 00:33:40,600 --> 00:33:43,160 ACTIVATED AND THE 25 MOST 919 00:33:43,160 --> 00:33:44,920 INACTIVATED. 920 00:33:44,920 --> 00:33:48,400 IMPORTANTLY, THESE ANALYSES 921 00:33:48,400 --> 00:33:49,320 IDENTIFY ONLY 407 MASTER 922 00:33:49,320 --> 00:33:52,720 VALIDATORS AND WE EVALUATED 141 923 00:33:52,720 --> 00:33:54,160 THAT ARE ACTUALLY SUBTYPE 924 00:33:54,160 --> 00:33:54,920 ESSENTIAL. 925 00:33:54,920 --> 00:33:55,240 35%. 926 00:33:55,240 --> 00:33:56,960 A PRETTY STRIKING ENRICHMENT. 927 00:33:56,960 --> 00:33:58,240 SO NOW I WANT TO TELL YOU HOW WE 928 00:33:58,240 --> 00:33:59,120 TARGET THESE THINGS IN THE 929 00:33:59,120 --> 00:34:01,880 CLINIC BEFORE SHOWING YOU HOW WE 930 00:34:01,880 --> 00:34:03,800 DO IT AT THE SINGLE CELL LEVEL. 931 00:34:03,800 --> 00:34:05,720 SO THIS IS A LARGE COLLABORATION 932 00:34:05,720 --> 00:34:08,600 THAT'S WITHIN GOING 933 00:34:08,600 --> 00:34:12,240 THAT'S BEEN GOING ON FOR SEVEN 934 00:34:12,240 --> 00:34:17,720 YEARS, AND NOW -- PURSUING IT. 935 00:34:17,720 --> 00:34:22,400 SO A THIS IS BASED ON TWO 936 00:34:22,400 --> 00:34:22,680 ALGORITHMS. 937 00:34:22,680 --> 00:34:24,680 THE FIRST ONE IS NEW YORK AND 938 00:34:24,680 --> 00:34:25,360 CALIFORNIA DEPARTMENT OF HEALTH 939 00:34:25,360 --> 00:34:26,640 APPROVED AND CLIA COMPLIANT. 940 00:34:26,640 --> 00:34:29,000 THE FIRST ONE IS A REALLY VERY 941 00:34:29,000 --> 00:34:33,240 SIMPLE MINDED ADAPTATION AND 942 00:34:33,240 --> 00:34:35,960 EXTENSION OF THE ONCOGENE 943 00:34:35,960 --> 00:34:37,440 ADDICTION POSTULATE THAT IS 944 00:34:37,440 --> 00:34:39,120 ACTUALLY TUMOR MUTATION 945 00:34:39,120 --> 00:34:39,480 AGNOSTIC. 946 00:34:39,480 --> 00:34:41,040 HOW IS IT AGNOSTIC? 947 00:34:41,040 --> 00:34:42,880 BECAUSE ALL WE KNEW IS RANKING 948 00:34:42,880 --> 00:34:44,440 ALL THE PROTEINS IN A TUMOR FROM 949 00:34:44,440 --> 00:34:46,640 THE MOST INACTIVATED TO THE MOST 950 00:34:46,640 --> 00:34:48,400 BARELY ACTIVATED AND THEN WE 951 00:34:48,400 --> 00:34:49,600 WALKED DOWN THIS LIST UNTIL WE 952 00:34:49,600 --> 00:34:54,280 HIT ONE FOR WHICH A HIGH 953 00:34:54,280 --> 00:34:55,680 AFFINITY PHARMACOLOGICAL 954 00:34:55,680 --> 00:34:58,000 INHIBITOR ALREADY EXISTS. 955 00:34:58,000 --> 00:35:00,320 SO ABOUT 20% OF METASTATIC 956 00:35:00,320 --> 00:35:07,720 BREAST CANCER WE IDENTIFY AS 957 00:35:07,720 --> 00:35:11,240 HDAC6 -- VERY HIGH INFINITY 958 00:35:11,240 --> 00:35:12,600 INHIBITOR SO WE STARTED CLINICAL 959 00:35:12,600 --> 00:35:16,040 TRIAL IN COLLABORATION WITH JOSE 960 00:35:16,040 --> 00:35:20,840 SILVA AND -- NOW AT ST. JUDE AND 961 00:35:20,840 --> 00:35:22,800 REALLY CAME FROM MY LAB, IN 962 00:35:22,800 --> 00:35:23,360 BREAST CANCER. 963 00:35:23,360 --> 00:35:25,160 WHAT YOU SEE HERE WAS REALLY 964 00:35:25,160 --> 00:35:27,120 QUITE STRIKING. 965 00:35:27,120 --> 00:35:28,360 BASICALLY EVERY SINGLE PATIENT 966 00:35:28,360 --> 00:35:30,640 THAT RESPONDED ACTUALLY HAD 967 00:35:30,640 --> 00:35:33,880 HIGHER -- ACTIVITY OF 968 00:35:33,880 --> 00:35:37,800 HDAC6 COMPUTED BY VIPER ANALYSIS 969 00:35:37,800 --> 00:35:40,080 COMPARED TO -- YOU CAN SEE 970 00:35:40,080 --> 00:35:41,800 SIGNIFICANT RESPONSE, RESPONDERS 971 00:35:41,800 --> 00:35:44,920 VERSUS NON-RESPONDERS AND THE -- 972 00:35:44,920 --> 00:35:50,600 CURVE WAS ESSENTIALLY 1. 973 00:35:50,600 --> 00:35:52,000 NOT THAT THE STUDY IS GOING TO 974 00:35:52,000 --> 00:35:54,360 SAVE PEOPLE'S LIVES NECESSARILY 975 00:35:54,360 --> 00:35:55,440 YOU SEE RECURRENCE BECAUSE 976 00:35:55,440 --> 00:35:56,880 YOU'RE TARGETING ONLY ONE 977 00:35:56,880 --> 00:35:57,840 PROTEIN, BUT AT LEAST IT TELLS 978 00:35:57,840 --> 00:35:59,360 YOU WE CAN PREDICT WITH VERY 979 00:35:59,360 --> 00:36:01,360 HIGH ACCURACY WHO WILL RESPOND 980 00:36:01,360 --> 00:36:05,040 CLINICALLY, NOT JUST IN A PDX 981 00:36:05,040 --> 00:36:06,160 MODEL OR IN A CELL LINE. 982 00:36:06,160 --> 00:36:09,560 SO THIS IS -- ARCHIVE AND IT'S 983 00:36:09,560 --> 00:36:10,520 IN REVIEW. 984 00:36:10,520 --> 00:36:13,840 SO THEN WE EXTENDED -- AND 985 00:36:13,840 --> 00:36:14,800 BASICALLY NOW WE'VE DONE THIS 986 00:36:14,800 --> 00:36:18,800 FOR EVERY COHORT NOT ONLY IN BCJ 987 00:36:18,800 --> 00:36:23,040 BUT TARGET -- WE CAN THINK OF, 988 00:36:23,040 --> 00:36:24,880 ESSENTIALLY WE RANK EVERY 989 00:36:24,880 --> 00:36:26,760 PROTEIN THAT IS DRUGGABLE -- IT 990 00:36:26,760 --> 00:36:28,200 IS THE MOST BARELY ACTIVATED 991 00:36:28,200 --> 00:36:28,400 ONE. 992 00:36:28,400 --> 00:36:31,800 SO FOR INSTANCE, WE FOUND THAT 993 00:36:31,800 --> 00:36:34,440 EXPLORING ONE WHICH IS A TARGET 994 00:36:34,440 --> 00:36:38,600 OF -- IS ACTUALLY REALLY BARELY 995 00:36:38,600 --> 00:36:41,640 ACTIVATED IN -- TUMORS AND GERM 996 00:36:41,640 --> 00:36:43,720 CELL TUMORS, IN FACT MORE SO 997 00:36:43,720 --> 00:36:45,160 THAN IN ANY OTHER TUMOR COHORT. 998 00:36:45,160 --> 00:36:47,200 SO YOU CAN SEE THE SENSITIVITY 999 00:36:47,200 --> 00:36:52,040 IS 10 TO MINUS 8 MICROMOLAR 1000 00:36:52,040 --> 00:36:53,400 RANGE, THESE TUMORS WHEN 1001 00:36:53,400 --> 00:36:54,480 TRANSPLANTED RESPONDED 1002 00:36:54,480 --> 00:36:56,800 STRIKINGLY WELL, EE STENGSLY 1003 00:36:56,800 --> 00:36:58,880 COMPLETE RESPONSE TO THE DRUG 1004 00:36:58,880 --> 00:37:00,440 AND THEN SHOWING RELAPSE WHEN 1005 00:37:00,440 --> 00:37:02,920 YOU STOPPED ADMINISTERING THE 1006 00:37:02,920 --> 00:37:03,120 DRUG. 1007 00:37:03,120 --> 00:37:04,280 AND SO THESE ARE PATIENTS THAT 1008 00:37:04,280 --> 00:37:07,600 HAVE BEEN TREATED BASED ON THESE 1009 00:37:07,600 --> 00:37:11,000 FINDINGS, THIS IS A 70-YEAR-OLD 1010 00:37:11,000 --> 00:37:14,000 PATIENT THAT HAD A RECURRENT 1011 00:37:14,000 --> 00:37:15,400 LUNG TUMOR ESSENTIALLY FAILED TO 1012 00:37:15,400 --> 00:37:18,680 RESPOND TO PRETTY MUCH ANY 1013 00:37:18,680 --> 00:37:20,400 THERAPY THROWN AT IT, 1014 00:37:20,400 --> 00:37:21,280 NUMEROUS -- IN THE PERITONEAL 1015 00:37:21,280 --> 00:37:23,440 CAVITY AND REFUSED TO GET 1016 00:37:23,440 --> 00:37:24,560 ADDITIONAL CHEMO TOXIC THERAPY 1017 00:37:24,560 --> 00:37:25,360 BECAUSE OF SIDE EFFECTS. 1018 00:37:25,360 --> 00:37:28,240 HE WAS PUT ON -- AND ESSENTIALLY 1019 00:37:28,240 --> 00:37:30,080 HAS BEEN DOING SPECTACULARLY 1020 00:37:30,080 --> 00:37:30,320 WELL. 1021 00:37:30,320 --> 00:37:32,200 THERE'S BEEN NO DETECTABLE 1022 00:37:32,200 --> 00:37:33,720 LESION AFTER HIS LAST SURGERY, 1023 00:37:33,720 --> 00:37:35,240 AND HE'S BACK IN SCHOOL AND 1024 00:37:35,240 --> 00:37:36,400 PLAYING SOCCER. 1025 00:37:36,400 --> 00:37:41,040 THIS IS AN ULTRA-RARE TUMOR. 1026 00:37:41,040 --> 00:37:42,280 THERE'S ONLY 40 REPORTED CASES 1027 00:37:42,280 --> 00:37:47,200 OF THIS TUMOR, NOT PER YEAR IN, IN 1028 00:37:47,200 --> 00:37:47,800 TOTAL. 1029 00:37:47,800 --> 00:37:49,360 A 14-YEAR-OLD PATIENT PRESENTED 1030 00:37:49,360 --> 00:37:50,440 WITH THIS TUMOR, FAILED TO 1031 00:37:50,440 --> 00:37:51,960 RESPOND TO EVERY POSSIBLE 1032 00:37:51,960 --> 00:37:53,760 THERAPY, INCLUDING A THERAPY 1033 00:37:53,760 --> 00:37:56,040 MEANT FOR -- TUMORS THAT DID 1034 00:37:56,040 --> 00:37:58,800 ABSOLUTELY NOTHING FOR HIM, AND 1035 00:37:58,800 --> 00:38:00,560 ONCOTARGET FOUND THAT PDJ -- 1036 00:38:00,560 --> 00:38:04,320 BETA WAS THE MOST -- ACTIVATED 1037 00:38:04,320 --> 00:38:05,200 PROTEIN AND -- ACTUALLY WELL 1038 00:38:05,200 --> 00:38:07,120 TOLERATED IN PATIENTS WAS THEN 1039 00:38:07,120 --> 00:38:08,520 SELECTED BY THE ONCOLOGIST TO 1040 00:38:08,520 --> 00:38:10,920 TREAT THE PATIENT AT MEMORIAL. 1041 00:38:10,920 --> 00:38:13,120 AND YOU CAN SEE REALLY THE 1042 00:38:13,120 --> 00:38:14,000 STRIKING RESPONSE AT THE LEVEL 1043 00:38:14,000 --> 00:38:17,000 OF THE IMAGING. 1044 00:38:17,000 --> 00:38:18,280 SO NOW LET ME TALK A LITTLE BIT 1045 00:38:18,280 --> 00:38:19,560 ABOUT THE MORE SOPHISTICATED 1046 00:38:19,560 --> 00:38:19,800 APPROACH. 1047 00:38:19,800 --> 00:38:23,080 SO I TOLD YOU THERE'S 25 MASTER 1048 00:38:23,080 --> 00:38:25,160 REGULATORS BARELY ACTIVATED, 25 1049 00:38:25,160 --> 00:38:26,840 THAT ARE INACTIVATED THAT 1050 00:38:26,840 --> 00:38:28,400 EXPLAIN THE GENETICS OF THE 1051 00:38:28,400 --> 00:38:28,600 SAMPLE. 1052 00:38:28,600 --> 00:38:30,400 WOULDN'T IT BE WONDERFUL IF WE 1053 00:38:30,400 --> 00:38:32,120 COULD FIND A DRUG THAT ACTUALLY 1054 00:38:32,120 --> 00:38:33,640 INVERT THEIR ACTIVITY, SO 1055 00:38:33,640 --> 00:38:36,080 BASICALLY CAN WE DO DRUG 1056 00:38:36,080 --> 00:38:39,120 PERTURBATION IN CELLS THAT 1057 00:38:39,120 --> 00:38:39,880 RECAPITULATE THE ACTIVITIES OF 1058 00:38:39,880 --> 00:38:40,960 THE MASTER REGULATORS WITH A 1059 00:38:40,960 --> 00:38:42,320 LARGE RANGE OF DRUGS AND THEN 1060 00:38:42,320 --> 00:38:44,040 FIND THE DRUG THAT IS ABLE TO 1061 00:38:44,040 --> 00:38:45,560 COMPLETELY SWITCH THE ACTIVITY. 1062 00:38:45,560 --> 00:38:47,520 NOW THIS SOUNDS INSANE BECAUSE 1063 00:38:47,520 --> 00:38:48,880 IT'S HARD ENOUGH TO FIND A DRUG 1064 00:38:48,880 --> 00:38:50,720 THAT TARGETS ONE PROTEIN, LET 1065 00:38:50,720 --> 00:38:52,240 ALONE 50, BUT THE REASON THIS 1066 00:38:52,240 --> 00:38:53,640 ACTUALLY WORKS AND WORKS QUITE 1067 00:38:53,640 --> 00:38:59,280 WELL, THIS IS A NATURE GENETICS 1068 00:38:59,280 --> 00:39:04,120 PAPER IN ENDOCRINE TUMORS OF THE 1069 00:39:04,120 --> 00:39:06,480 GUT, IS THAT AS I SHOWED YOU FOR 1070 00:39:06,480 --> 00:39:07,880 PANCREATIC CANCER, THESE MASTER 1071 00:39:07,880 --> 00:39:13,000 REGULATORS REALLY WORK AS A MOD AS A 1072 00:39:13,000 --> 00:39:14,120 MODULE, SO WHEN YOU HIT THE 1073 00:39:14,120 --> 00:39:15,720 RIGHT ENTRY POINT EVEN IN A WAY 1074 00:39:15,720 --> 00:39:17,600 YOU DON'T NECESSARILY 1075 00:39:17,600 --> 00:39:19,440 UNDERSTAND, THE ENTIRE MODULE 1076 00:39:19,440 --> 00:39:20,320 ACTUALLY SWITCHES OFF. 1077 00:39:20,320 --> 00:39:21,400 THAT'S WHAT CONTROLS THE STATE 1078 00:39:21,400 --> 00:39:23,760 OF THE CELL SO YOU EXPECT EERD 1079 00:39:23,760 --> 00:39:24,960 THE TUMOR TO EITHER REPROGRAM OR 1080 00:39:24,960 --> 00:39:26,200 TO DIE. 1081 00:39:26,200 --> 00:39:27,720 IT'S VERY IMPORTANT TO SELECT 1082 00:39:27,720 --> 00:39:29,040 MODELS THAT ARE ACTUALLY MATCHED 1083 00:39:29,040 --> 00:39:30,760 TO THE MASTER REGULATORS 1084 00:39:30,760 --> 00:39:32,240 OTHERWISE YOU GET COMPLETE JUNK 1085 00:39:32,240 --> 00:39:34,120 BUT WE'VE SHOWN THIS IN SEVERAL 1086 00:39:34,120 --> 00:39:38,800 PAPERS THAT WEECH WE'VE PUBLISHED. 1087 00:39:38,800 --> 00:39:42,720 SO THESE ARE GENOME-WIDE IN 1088 00:39:42,720 --> 00:39:45,360 COMPARISON TO CMAP, THEY'RE NOT 1089 00:39:45,360 --> 00:39:48,840 USING 1,000 -- BUT THEY'RE -- 1090 00:39:48,840 --> 00:39:53,120 THEY'RE DYING CELL LINES, BASED 1091 00:39:53,120 --> 00:39:54,800 ON CONSERVATION OF MASTER 1092 00:39:54,800 --> 00:39:56,320 REGULATORS, AND THE PERTURBATION 1093 00:39:56,320 --> 00:39:58,440 WAS DONE AT THE CONCENTRATION OF 1094 00:39:58,440 --> 00:40:00,840 THE 48-HOUR IC20 CONCENTRATIONS, 1095 00:40:00,840 --> 00:40:02,880 BASICALLY THE HIGHEST SUBLETHAL 1096 00:40:02,880 --> 00:40:04,120 CONCENTRATION OF THE DRUG YOU 1097 00:40:04,120 --> 00:40:06,160 COULD ACHIEVE, SO THAT YOU DON'T 1098 00:40:06,160 --> 00:40:07,480 HAVE CONFOUNDING FACTOR, 1099 00:40:07,480 --> 00:40:09,880 CONFOUNDING EVENT FROM CELL 1100 00:40:09,880 --> 00:40:11,440 STRESS PATHWAYS OR CELL DEATH 1101 00:40:11,440 --> 00:40:13,600 PATHWAYS. 1102 00:40:13,600 --> 00:40:16,360 SO THIS REPERTOIRE CAN BE NOW 1103 00:40:16,360 --> 00:40:19,280 USED TO MAKE ONCOTREAT 1104 00:40:19,280 --> 00:40:20,760 PREDICTIONS IN VIRTUALLY ANY 1105 00:40:20,760 --> 00:40:21,000 PATIENT. 1106 00:40:21,000 --> 00:40:24,400 SO THIS IS THE N OF 1 STUDY, 1107 00:40:24,400 --> 00:40:25,920 AGAIN SEVEN YEARS IN THE WORKS, 1108 00:40:25,920 --> 00:40:28,640 WHERE WE ENROLLED ABOUT 130 1109 00:40:28,640 --> 00:40:31,560 PATIENTS ACROSS 18 DIFFERENT 1110 00:40:31,560 --> 00:40:37,320 MALIGNANCY, GENERATED 45 PDX 1111 00:40:37,320 --> 00:40:38,920 MODELS, 34 DIFFERENT DRUGS 1112 00:40:38,920 --> 00:40:40,520 PREDICTED BY EITHER ONCOTREAT OR 1113 00:40:40,520 --> 00:40:41,160 CONOCO TARGET. 1114 00:40:41,160 --> 00:40:42,480 SO THESE DRUGS WERE PREDICTED 1115 00:40:42,480 --> 00:40:44,160 EITHER BECAUSE THEY WERE 1116 00:40:44,160 --> 00:40:45,600 TARGETING A SINGLE MASTER 1117 00:40:45,600 --> 00:40:49,600 REGULATOR OR THEY WERE 1118 00:40:49,600 --> 00:40:50,200 TARGETING -- SIGNATURE. 1119 00:40:50,200 --> 00:40:52,200 SO I THINK THESE ARE PRETTY 1120 00:40:52,200 --> 00:40:52,720 STRIKING RESULTS. 1121 00:40:52,720 --> 00:40:54,160 YOU'VE SEEN THAT THESE TUMORS 1122 00:40:54,160 --> 00:40:57,960 WERE SELECTED SPECIFICALLY 1123 00:40:57,960 --> 00:40:59,440 ENROLLMENT CRITERIA TO HAVE 1124 00:40:59,440 --> 00:41:01,680 FAILED THREE TO SEVEN LINES OF 1125 00:41:01,680 --> 00:41:02,440 THERAPY, ESSENTIALLY CONSIDERED 1126 00:41:02,440 --> 00:41:03,560 TO BE UNTREATABLE. 1127 00:41:03,560 --> 00:41:07,320 YOU CAN SEE ALL OF THEM DOUBLE 1128 00:41:07,320 --> 00:41:08,640 IN VOLUME SIZE IN LESS THAN 30 1129 00:41:08,640 --> 00:41:10,160 DAYS. 1130 00:41:10,160 --> 00:41:12,600 HOWEVER, IN 16 ONCOTARGET DRUG 1131 00:41:12,600 --> 00:41:15,080 ARMS AND 18 ONCOTREAT DRUG ARMS, 1132 00:41:15,080 --> 00:41:17,720 EIGHT OF WHICH ARE PREDICTED BY 1133 00:41:17,720 --> 00:41:18,040 METHODOLOGIES. 1134 00:41:18,040 --> 00:41:20,080 YOU CAN SEE ONCOTARGET INDUCED A 1135 00:41:20,080 --> 00:41:24,640 RESPONSE IN ABOUT 60% OR MORE OF 1136 00:41:24,640 --> 00:41:27,000 THESE PDS MODELS, AND ONCOTREAT, 1137 00:41:27,000 --> 00:41:27,960 MORE 90%. 1138 00:41:27,960 --> 00:41:30,280 SO THIS WAS VERY ENCOURAGING, 1139 00:41:30,280 --> 00:41:31,920 BUT WE ALSO WANT TO KNOW, WHAT 1140 00:41:31,920 --> 00:41:34,400 ABOUT USING CONVENTIONAL THERAPY 1141 00:41:34,400 --> 00:41:37,400 AS OPPOSED TO LETHAL CONTROL. 1142 00:41:37,400 --> 00:41:41,600 SO WE DID A TEST WHERE WE 1143 00:41:41,600 --> 00:41:42,600 RANDOMLY -- COMPARED THEM TO 1144 00:41:42,600 --> 00:41:44,000 THE -- YOU CAN SEE HERE THERE'S 1145 00:41:44,000 --> 00:41:45,040 ABSOLUTELY NO RESPONSE. 1146 00:41:45,040 --> 00:41:47,000 AND THIS WAS DONE IN 13 NEGATIVE 1147 00:41:47,000 --> 00:41:49,880 CONTROL ARMS, SO THIS IN OUR 1148 00:41:49,880 --> 00:41:51,560 OPINION IS THE FIRST STUDY WHERE 1149 00:41:51,560 --> 00:41:53,640 THE RESULTS OF THE STUDY THAT 1150 00:41:53,640 --> 00:41:54,400 PREDICTS ACTIVITY AND 1151 00:41:54,400 --> 00:41:57,080 SENSITIVITY OF A DRUG IN VIVO, 1152 00:41:57,080 --> 00:41:58,080 DE NOVO, IS ACTUALLY VALIDATED 1153 00:41:58,080 --> 00:42:00,440 IN A WAY THAT IS STATISTICALLY 1154 00:42:00,440 --> 00:42:00,760 SIGNIFICANT. 1155 00:42:00,760 --> 00:42:03,640 AND YOU CAN SEE HERE PANCREATIC 1156 00:42:03,640 --> 00:42:05,040 CANCER, THESE ARE THE FIVE DRUGS 1157 00:42:05,040 --> 00:42:06,080 THAT WE PREDICTED AND YOU CAN 1158 00:42:06,080 --> 00:42:08,240 SEE THAT THREE OF THEM ACTUALLY 1159 00:42:08,240 --> 00:42:11,480 HAD PRETTY SIGNIFICANT EFFECT, 1160 00:42:11,480 --> 00:42:12,320 PARTIAL RESPONSE OR STABLE 1161 00:42:12,320 --> 00:42:12,600 DISEASE. 1162 00:42:12,600 --> 00:42:14,920 ONE OF THEM WAS BORDERLINE, ONLY 1163 00:42:14,920 --> 00:42:17,240 ONE FAILED TO RESPOND IN A 1164 00:42:17,240 --> 00:42:17,640 MOUSE. 1165 00:42:17,640 --> 00:42:18,920 THE MORE INTERESTING THING IS 1166 00:42:18,920 --> 00:42:21,600 THAT BY TAKING SAMPLES OF THREE 1167 00:42:21,600 --> 00:42:23,400 HOURS AFTER THE THIRD DOSE OF 1168 00:42:23,400 --> 00:42:25,960 THE DRUGS WHICH ARE BASICALLY 1169 00:42:25,960 --> 00:42:27,280 PHARMACODYNAMIC SAMPLES, WE 1170 00:42:27,280 --> 00:42:29,400 COULD VALIDATE THAT IN EVERY 1171 00:42:29,400 --> 00:42:31,040 SINGLE CASE OUT OF THE 18 DRUGS 1172 00:42:31,040 --> 00:42:36,440 WE TESTED WE DON'T TREAT, SO 15 1173 00:42:36,440 --> 00:42:40,040 OUT OF 18 TREATMENT EXACTLY 1174 00:42:40,040 --> 00:42:41,440 REVERSE -- IN VIVO THAT WE HAD 1175 00:42:41,440 --> 00:42:42,120 PREDICTED FROM CELL LINES. 1176 00:42:42,120 --> 00:42:43,000 SO THIS IS VERY IMPORTANT 1177 00:42:43,000 --> 00:42:44,200 BECAUSE IT VALIDATES THE FACT 1178 00:42:44,200 --> 00:42:46,840 THAT WHEN WE MATCH THESE CELL 1179 00:42:46,840 --> 00:42:48,400 LINES WITH THE ONCOMATCH, WE 1180 00:42:48,400 --> 00:42:50,120 ACTUALLY FIND MODELS THAT ARE 1181 00:42:50,120 --> 00:42:51,360 RELEVANT TO THE IN VIVO STUDY 1182 00:42:51,360 --> 00:42:53,880 AND WE DON'T CARE AT ALL WHETHER 1183 00:42:53,880 --> 00:42:56,040 THE DRUGS WILL KILL THE CELL 1184 00:42:56,040 --> 00:42:58,040 LINES BECAUSE THAT'S NOT 1185 00:42:58,040 --> 00:43:00,040 HAPPENING, BUT WE CARE ABOUT 1186 00:43:00,040 --> 00:43:02,000 UNDERSTANDING THE DRUG MECHANISM 1187 00:43:02,000 --> 00:43:05,440 OF ACTION BECAUSE THEN WE CAN 1188 00:43:05,440 --> 00:43:07,000 MATCH TO THE -- OF THE TUMOR IN 1189 00:43:07,000 --> 00:43:07,920 THE PATIENT. 1190 00:43:07,920 --> 00:43:09,280 SO LET ME SHOW YOU HOW THIS CAN 1191 00:43:09,280 --> 00:43:11,360 BE MOVED IN SINGLE CELLS. 1192 00:43:11,360 --> 00:43:15,840 WE STUDIED THIS IN COLLABORATION 1193 00:43:15,840 --> 00:43:17,960 WITH JERRY -- NOW BMS AND 1194 00:43:17,960 --> 00:43:19,640 SEVERAL OTHER INVESTIGATORS BY 1195 00:43:19,640 --> 00:43:22,000 FOCUSING ON A POPULATION OF 1196 00:43:22,000 --> 00:43:23,240 TUMOR AND SHADING CELLS THAT CAN 1197 00:43:23,240 --> 00:43:27,000 BE ENRICHED FOR BY SORTING USING 1198 00:43:27,000 --> 00:43:29,040 CD49F AND -- THIS POPULATION, 1199 00:43:29,040 --> 00:43:32,160 SOME PEOPLE CALL IT ENRICH FOR 1200 00:43:32,160 --> 00:43:34,400 CANCER STEM CELLS, WE DON'T LIKE 1201 00:43:34,400 --> 00:43:40,680 THAT TERMINOLOGY, SO WE STARTED 1202 00:43:40,680 --> 00:43:48,920 ABOUT 75% -- AND TWO TRIPLE 1203 00:43:48,920 --> 00:43:53,480 NEGATIVE AND 25% WE BROADLY DID 1204 00:43:53,480 --> 00:43:55,000 EPCAM. 1205 00:43:55,000 --> 00:43:56,760 WE DID -- WE PREDICTED DRUGS FOR 1206 00:43:56,760 --> 00:43:58,200 THE DIFFERENT SUBTYPES WE 1207 00:43:58,200 --> 00:44:00,400 IDENTIFIED, TREAT IT IN VIVO AND 1208 00:44:00,400 --> 00:44:05,200 THEN AVEST SES ASSESS IF THE DRUG DID WHA T 1209 00:44:05,200 --> 00:44:06,800 IT WAS SUPPOSED TO DO. 1210 00:44:06,800 --> 00:44:08,040 YOU SEE HERE, IT'S REALLY 1211 00:44:08,040 --> 00:44:09,200 STRIKING BECAUSE FIRST OF ALL, 1212 00:44:09,200 --> 00:44:11,960 IF YOU DO GENE EXPRESSION BASED 1213 00:44:11,960 --> 00:44:13,960 CLUSTERING, YOU SEE EVERY 1214 00:44:13,960 --> 00:44:14,800 PORTION FORMS A DIFFERENT 1215 00:44:14,800 --> 00:44:15,320 CLUSTER. 1216 00:44:15,320 --> 00:44:16,560 SO PEOPLE CALL THIS A BATCH 1217 00:44:16,560 --> 00:44:17,080 EFFECT. 1218 00:44:17,080 --> 00:44:19,520 THIS IS NOT BATCH EFFECT. 1219 00:44:19,520 --> 00:44:20,400 THIS IS BIOLOGY. 1220 00:44:20,400 --> 00:44:22,360 THE BIOLOGY COMES FROM THE LARGE 1221 00:44:22,360 --> 00:44:25,720 COPY NUMBER ALTERATION THAT -- 1222 00:44:25,720 --> 00:44:26,720 INDIVIDUAL PATIENT WHICH 1223 00:44:26,720 --> 00:44:28,400 CONTRIBUTES TO SOME DIFFERENCE 1224 00:44:28,400 --> 00:44:34,240 IN THE -- THE CELL BUFFERS THOSE 1225 00:44:34,240 --> 00:44:35,320 DIFFERENCES SO THE DIFFERENCE 1226 00:44:35,320 --> 00:44:36,400 THAT'S IMPORTANT IN ESTABLISHING 1227 00:44:36,400 --> 00:44:37,560 THE STATE OF THE CELL AS 1228 00:44:37,560 --> 00:44:38,760 TRANSFORMED CELLS ARE RETRAINED. 1229 00:44:38,760 --> 00:44:40,400 SO WHAT HAPPENS IS WHEN YOU GO 1230 00:44:40,400 --> 00:44:43,600 FROM GENE ECK PRETION TO VIPER 1231 00:44:43,600 --> 00:44:44,840 ACTIVITY, NOW ALL THE PATIENT 1232 00:44:44,840 --> 00:44:45,360 OVERLAP. 1233 00:44:45,360 --> 00:44:47,280 AND THIS REQUIRES NO 1234 00:44:47,280 --> 00:44:48,280 NORMALIZATION, NO BATCH EFFECT 1235 00:44:48,280 --> 00:44:48,680 CORRECTION. 1236 00:44:48,680 --> 00:44:50,000 AND WHAT YOU SEE NOW IS THE 1237 00:44:50,000 --> 00:44:50,920 DIFFERENCE BETWEEN THE TOP AND 1238 00:44:50,920 --> 00:44:54,000 THE BOTTOM, IS THE HORMONE 1239 00:44:54,000 --> 00:44:55,200 POSITIVE VERSUS THE TRIPLE 1240 00:44:55,200 --> 00:44:55,760 NEGATIVE. 1241 00:44:55,760 --> 00:44:57,200 SO BASICALLY YOU'RE RETAINING 1242 00:44:57,200 --> 00:44:58,240 IMPORTANT BIOLOGY AND YOU'RE 1243 00:44:58,240 --> 00:45:00,800 MISSING OUT THE THINGS THAT ARE 1244 00:45:00,800 --> 00:45:02,800 JUST OBTURATION THAT ARE NOT 1245 00:45:02,800 --> 00:45:03,560 FUNCTIONALLY RELEVANT. 1246 00:45:03,560 --> 00:45:04,840 AND ALSO YOU SEE THAT WHEN YOU 1247 00:45:04,840 --> 00:45:06,840 GO LEFT TO RIGHT, YOU SEE 1248 00:45:06,840 --> 00:45:08,680 STRIKING ENRICHMENT IN MARKERS 1249 00:45:08,680 --> 00:45:11,040 THAT HAVE BEEN PUBLISHED AS 1250 00:45:11,040 --> 00:45:17,360 ESTABLISHED MARKERS OF 1251 00:45:17,360 --> 00:45:21,120 STEMNESS -- IN FACT, BY 1252 00:45:21,120 --> 00:45:25,280 CYTOTRACE ANALYSIS, YOU CAN SEE 1253 00:45:25,280 --> 00:45:27,160 STRIKINGLY HIGH -- SO WHAT WE 1254 00:45:27,160 --> 00:45:30,440 DID, WE TOOK THE 10 MOST STEM 1255 00:45:30,440 --> 00:45:31,800 LIKE CELLS FROM EACH PATIENT AND 1256 00:45:31,800 --> 00:45:33,560 COMPARED THEM TO THE 10 MOST 1257 00:45:33,560 --> 00:45:34,680 DIFFERENTIATING CELLS FROM THE 1258 00:45:34,680 --> 00:45:36,400 SAME PATIENT AND WE DID THIS 1259 00:45:36,400 --> 00:45:37,240 INDEPENDENTLY ACROSS EACH 1260 00:45:37,240 --> 00:45:39,240 PATIENT, AND THEN WE INTEGRATED 1261 00:45:39,240 --> 00:45:41,640 THE RESULTS. 1262 00:45:41,640 --> 00:45:43,480 THE MASTER REGULATORS WERE 1263 00:45:43,480 --> 00:45:44,480 EXACTLY THE SAME ACROSS THE 1264 00:45:44,480 --> 00:45:46,320 PATIENTS SO THE INTEGRATION WAS 1265 00:45:46,320 --> 00:45:50,320 VERY, VERY SUCCESSFUL, THEN WE 1266 00:45:50,320 --> 00:45:51,440 USED ONCOTREAT PO TREE 1267 00:45:51,440 --> 00:45:53,000 DICTIONARY -- WHICH REALLY HAD A 1268 00:45:53,000 --> 00:45:54,160 STRONG CONSERVATION FOR THE 1269 00:45:54,160 --> 00:45:55,440 ACTIVITY OF THESE MASTER 1270 00:45:55,440 --> 00:45:55,880 REGULATORS. 1271 00:45:55,880 --> 00:45:59,920 AND WHAT WAS SURPRISING IS THAT 1272 00:45:59,920 --> 00:46:03,440 TWO DRUGS, THESE ARE ANTE -- 1273 00:46:03,440 --> 00:46:04,840 TYPICALLY NEVER USED IN CANCER 1274 00:46:04,840 --> 00:46:09,960 BUT CAME UP REALLY -- IS THE 1275 00:46:09,960 --> 00:46:11,760 NUMBER ONE MOST STATISTICALLY 1276 00:46:11,760 --> 00:46:12,400 SIGNIFICANT. 1277 00:46:12,400 --> 00:46:14,680 THESE MICE TRANSPLANTED WITH -- 1278 00:46:14,680 --> 00:46:16,920 THE TUMOR BIOPSY FOR A TRIPLE 1279 00:46:16,920 --> 00:46:20,800 NEGATIVE, VERY AGGRESSIVE 1280 00:46:20,800 --> 00:46:24,400 PATIENT, WITH -- THEY PROFILED 1281 00:46:24,400 --> 00:46:25,840 THEM AND SEQUENCED THEM, AND 1282 00:46:25,840 --> 00:46:27,040 WHAT YOU'RE SEEING HERE IS THAT 1283 00:46:27,040 --> 00:46:30,400 WE MEASURE THE ENRICHMENT 1284 00:46:30,400 --> 00:46:32,400 INCREASING DENSITY BY COMPARING 1285 00:46:32,400 --> 00:46:34,360 EACH TREATMENT TO THE -- 1286 00:46:34,360 --> 00:46:34,800 CONTROL. 1287 00:46:34,800 --> 00:46:37,560 SO AS YOU EXPECT WHEN YOU TREAT 1288 00:46:37,560 --> 00:46:38,800 WITH PACLITAXEL WHICH IS KNOWN 1289 00:46:38,800 --> 00:46:41,160 TO TARGET DIFFERENTIATED 1290 00:46:41,160 --> 00:46:42,760 PROLIFERATIVE CELLS, YOU SEE 1291 00:46:42,760 --> 00:46:44,600 STRIKING DEPLETION OF THE CELLS 1292 00:46:44,600 --> 00:46:46,680 WHICH ARE IN BLUE, THE COOLER 1293 00:46:46,680 --> 00:46:53,360 TONES OF THE COLOR SPECTRUM, AND 1294 00:46:53,360 --> 00:46:56,160 DRAMATIC REPLENISHMENT OF THE 1295 00:46:56,160 --> 00:46:56,800 STEM-LIKE COMPARTMENT. 1296 00:46:56,800 --> 00:46:58,160 SO THESE TUMORS STOP GROWING 1297 00:46:58,160 --> 00:46:59,200 BECAUSE YOU'RE KILLING A LARGE 1298 00:46:59,200 --> 00:47:00,560 AMOUNT OF THE CELLS, BUT THEY 1299 00:47:00,560 --> 00:47:01,840 ACCUMULATE A LOT OF CELLS THAT 1300 00:47:01,840 --> 00:47:05,240 CAN RESTART AND REMOVE THE 1301 00:47:05,240 --> 00:47:06,560 TUMOR. 1302 00:47:06,560 --> 00:47:09,080 HOWEVER, WITH IVERMECTIN, 1303 00:47:09,080 --> 00:47:14,160 DRAMATICALLY BETTER WITH 1304 00:47:14,160 --> 00:47:14,880 ALBENDAZOLE -- MORE STEM LIKE 1305 00:47:14,880 --> 00:47:16,840 AND IN FACT INCREASE IN THE 1306 00:47:16,840 --> 00:47:19,440 CELLS IN THE POPULATION 1307 00:47:19,440 --> 00:47:20,880 DIFFERENTIATED AND PROLIFERATING 1308 00:47:20,880 --> 00:47:21,080 CELLS. 1309 00:47:21,080 --> 00:47:23,040 SO WE THOUGHT WE SHOULD USE 1310 00:47:23,040 --> 00:47:24,680 THESE TWO DRUGS IN COMBINATION, 1311 00:47:24,680 --> 00:47:27,400 MAYBE THEY SYNERGIZE. 1312 00:47:27,400 --> 00:47:29,240 INDEED WE DID THIS CLINICAL 1313 00:47:29,240 --> 00:47:36,400 STUDY, YOU CAN SEE THE ALBEN DA 1314 00:47:36,400 --> 00:47:39,520 SOL 1315 00:47:39,520 --> 00:47:40,720 ALBENDAZOLE DOES ABSOLUTELY 1316 00:47:40,720 --> 00:47:41,200 NOTHING ON ITS OWN. 1317 00:47:41,200 --> 00:47:44,240 WHEN YOU ACTUALLY TREATED WITH 1318 00:47:44,240 --> 00:47:45,480 COMBINATION, YOU GOT REGRESSION 1319 00:47:45,480 --> 00:47:48,800 AND THEN THE TUMOR WORKS 1320 00:47:48,800 --> 00:47:51,600 ESSENTIALLY TO A AN ESSENTIAL 1321 00:47:51,600 --> 00:47:52,040 STATE. 1322 00:47:52,040 --> 00:47:54,000 WE HAD TO STOP THIS CURVE HERE 1323 00:47:54,000 --> 00:47:55,480 BECAUSE THE TUMORS STARTED TO 1324 00:47:55,480 --> 00:47:58,680 GROW SO LARGE THAT SOME OF 1325 00:47:58,680 --> 00:47:59,960 THE -- STARTING TO HAVE TO BE 1326 00:47:59,960 --> 00:48:01,400 SACRIFICED SO WE COULDN'T 1327 00:48:01,400 --> 00:48:02,800 REPRESENT ALL THE MICE. 1328 00:48:02,800 --> 00:48:04,840 BUT IF YOU LOOK AT THE SPIDER 1329 00:48:04,840 --> 00:48:06,720 PLOTS, YOU NOW SEE THE MICE THAT 1330 00:48:06,720 --> 00:48:08,400 WERE ON THE COME BOW, FOUR OF 1331 00:48:08,400 --> 00:48:09,920 THE SIX HAD REALLY STRIKING 1332 00:48:09,920 --> 00:48:12,440 RESPONSE, WERE VERY DURABLE, AND 1333 00:48:12,440 --> 00:48:16,000 IN FACT THESE MICE WERE NOT -- 1334 00:48:16,000 --> 00:48:17,160 BUT RATHER BECAUSE THEY STARTED 1335 00:48:17,160 --> 00:48:17,440 TO ULS RATE. 1336 00:48:17,440 --> 00:48:19,720 SO WE'RE NOW REPEATING THE STUDY 1337 00:48:19,720 --> 00:48:28,400 BY FIRST TREATING WITH PACK 1338 00:48:28,400 --> 00:48:29,920 PACLITAXEL, NOW FIRST -- WE TAKE 1339 00:48:29,920 --> 00:48:31,200 DOWN THE TUMOR VOLUME BEFORE IT 1340 00:48:31,200 --> 00:48:35,800 HAS A CHANCE TO DO TOO MUCH, AND 1341 00:48:35,800 --> 00:48:37,400 THEN ALBEN DA ALBENDAZOLE. 1342 00:48:37,400 --> 00:48:38,840 I WANT TO CONCLUDE IN THE NEXT 1343 00:48:38,840 --> 00:48:39,800 THREE MINUTES WITH SOMETHING WE 1344 00:48:39,800 --> 00:48:41,520 CONSIDER TO BE A REALLY EXCITING 1345 00:48:41,520 --> 00:48:42,480 STUDY BECAUSE INSTEAD OF 1346 00:48:42,480 --> 00:48:44,080 TARGETING ACTUAL MORE CELLS, 1347 00:48:44,080 --> 00:48:46,600 WE'RE NOW TARGETING HIGHLY 1348 00:48:46,600 --> 00:48:47,760 IMMUNOSUPPRESSIVE CELLS IN THE 1349 00:48:47,760 --> 00:48:51,360 TUMOR MICRO ENENVIRONMENT. 1350 00:48:51,360 --> 00:48:57,640 SO WE ISOLATED BOTH PRIF LAL PERIPHERAL 1351 00:48:57,640 --> 00:48:59,200 AND TUMOR -- IN FOUR DIFFERENT 1352 00:48:59,200 --> 00:48:59,520 TUMOR TYPES. 1353 00:48:59,520 --> 00:49:02,800 WE ASKED CAN WE SEPARATE THE 1354 00:49:02,800 --> 00:49:05,120 INFILTRATING VERSUS 1355 00:49:05,120 --> 00:49:07,040 NON-INFILTRATING T-REGS BY GENE 1356 00:49:07,040 --> 00:49:09,200 EXPRESSION PROFILE, NEITHER BY 1357 00:49:09,200 --> 00:49:12,400 TUMOR TYPE OR ACROSS ALL FOUR 1358 00:49:12,400 --> 00:49:13,080 TUMORS. 1359 00:49:13,080 --> 00:49:17,840 HOWEVER, NOW YOU FIND 15 MASTER 1360 00:49:17,840 --> 00:49:18,960 REGULATORS -- VERSUS THE 1361 00:49:18,960 --> 00:49:23,360 PERIPHERAL T REGS AND ADDITIONAL 1362 00:49:23,360 --> 00:49:25,880 TWO -- IN COMMON THAT ARE 1363 00:49:25,880 --> 00:49:27,040 ACTUALLY ALSO SPECIFICALLY 1364 00:49:27,040 --> 00:49:28,600 ACTIVATED IN THE INFILTRATING T 1365 00:49:28,600 --> 00:49:31,840 REGS BUT NOT IN THE NON-T REG 1366 00:49:31,840 --> 00:49:33,360 INFILTRATING T-CELLS WHICH -- 1367 00:49:33,360 --> 00:49:35,120 YOU WANT TO MAINTAIN BECAUSE 1368 00:49:35,120 --> 00:49:38,080 OTHERWISE YOU LOSE THE -- 1369 00:49:38,080 --> 00:49:39,960 RESPONSE IN THE TUMOR. 1370 00:49:39,960 --> 00:49:42,560 SO TO VALIDATE THIS, WE DID A 1371 00:49:42,560 --> 00:49:45,680 CHIMERIC ASSAY WHERE YOU DO 1372 00:49:45,680 --> 00:49:48,400 ESSENTIALLY A FOOL CRISPR 1373 00:49:48,400 --> 00:49:53,680 KNOCKOUT SCREEN THAT -- IN 1374 00:49:53,680 --> 00:49:55,080 HEMATOPOIETIC STEM CELLS AND USE 1375 00:49:55,080 --> 00:49:57,040 THEM TO REMOP LATE THE BONE 1376 00:49:57,040 --> 00:50:00,200 MARROW OF A MOUSE, THEN AFTER 1377 00:50:00,200 --> 00:50:01,680 ABOUT 10 WEEKS WHERE THE IMMUNE 1378 00:50:01,680 --> 00:50:02,960 SYSTEM IS COMPLETELY REBOOTED 1379 00:50:02,960 --> 00:50:07,560 AND NOW EVERY CELL AND ABOUT 1380 00:50:07,560 --> 00:50:09,400 35% -- KNOCK OUT ONE OF THESE 1381 00:50:09,400 --> 00:50:12,400 PROTEINS, YOU NOW IMPLANT -- 1382 00:50:12,400 --> 00:50:13,560 VERY AGGRESSIVE COLON CANCER 1383 00:50:13,560 --> 00:50:14,600 MODEL, THEN YOU LOOK AT WHETHER 1384 00:50:14,600 --> 00:50:15,640 THERE WAS A DIFFERENCE IN 1385 00:50:15,640 --> 00:50:17,400 PERIPHERAL VERSUS TUMOR -- 1386 00:50:17,400 --> 00:50:20,400 CELLS, SO WE DID THIS IN TWO 1387 00:50:20,400 --> 00:50:21,600 DIFFERENT COURSES, THE SECOND 1388 00:50:21,600 --> 00:50:23,520 ONE, FILTERED WITH A REPORTER 1389 00:50:23,520 --> 00:50:24,360 FROM THE FIRST -- AND THEN 1390 00:50:24,360 --> 00:50:26,200 REPLANTED AND YOU CAN SEE 1391 00:50:26,200 --> 00:50:26,800 STRIKING CORRELATION BETWEEN THE 1392 00:50:26,800 --> 00:50:30,160 RESULTS IN TERMS OF THE PROFILE 1393 00:50:30,160 --> 00:50:31,480 OF REPRESENTATION OF THE 1394 00:50:31,480 --> 00:50:37,160 DIFFERENT MASTER REGULATORS GENE 1395 00:50:37,160 --> 00:50:38,320 TARGETING. 1396 00:50:38,320 --> 00:50:38,760 THE BARCODES. 1397 00:50:38,760 --> 00:50:43,520 AND THEN YOU CAN SEE HERE THIS 1398 00:50:43,520 --> 00:50:47,840 AXIS IS EXTRAORDINARY -- THE 1399 00:50:47,840 --> 00:50:52,520 TOP -- STATISTICAL 1400 00:50:52,520 --> 00:50:54,880 SIGNIFICANT -- 10 TO THE 170 IS 1401 00:50:54,880 --> 00:50:56,160 SIGNIFICANT, CAN YOU SEE THEY 1402 00:50:56,160 --> 00:50:57,960 ARE SIGNIFICANT BOTH IN TERMS OF 1403 00:50:57,960 --> 00:51:00,920 PERIPHERAL T REGS WHICH WERE 1404 00:51:00,920 --> 00:51:01,560 SELECTED FROM THE SPLEEN AND 1405 00:51:01,560 --> 00:51:06,800 ALSO WITH RESPECT TO NON-T REG 1406 00:51:06,800 --> 00:51:07,680 CD4 T-CELLS FROM THE TUMOR 1407 00:51:07,680 --> 00:51:08,160 MICROENVIRONMENT. 1408 00:51:08,160 --> 00:51:09,920 SO WE REPEATED THE ASSAY BUT 1409 00:51:09,920 --> 00:51:13,080 THIS TIME WE ONLY SILENCED TRPS1 WHICH IS A GENE THAT 1410 00:51:13,080 --> 00:51:14,240 NOBODY KNOWS ANYTHING ABOUT IT 1411 00:51:14,240 --> 00:51:15,800 IN RELATIONSHIP TO T REGS AND 1412 00:51:15,800 --> 00:51:17,200 YOU CAN SEE THAT'S QUITE A 1413 00:51:17,200 --> 00:51:19,920 STRIKING EFFECT IN TERMS OF 1414 00:51:19,920 --> 00:51:21,200 TUMOR GROWTH IN THE FIVE MODELS 1415 00:51:21,200 --> 00:51:23,720 WITH THE NON-TARGETING CONTROLS, 1416 00:51:23,720 --> 00:51:26,120 AND SIX MODELS WITH TARGETING 1417 00:51:26,120 --> 00:51:26,320 PRPS1. 1418 00:51:26,320 --> 00:51:31,400 IN FACT IF YOU LOOK AT A SINGLE 1419 00:51:31,400 --> 00:51:33,960 MOUSE, THREE OF THE MICE -- 1420 00:51:33,960 --> 00:51:35,440 WITHOUT ANY DRUG. 1421 00:51:35,440 --> 00:51:36,320 THIS IS A DIFFERENT MODEL 1422 00:51:36,320 --> 00:51:37,320 BECAUSE WE WANTED TO MAKE SURE 1423 00:51:37,320 --> 00:51:38,840 THIS WAS NOT SPECIFIC TO THAT 1424 00:51:38,840 --> 00:51:42,000 COLON CANCER MODEL, THIS IS A 1425 00:51:42,000 --> 00:51:45,400 HIGHLY IMMUNE RESISTANT AND 1426 00:51:45,400 --> 00:51:46,000 AGGRESSIVE SARCOMA. 1427 00:51:46,000 --> 00:51:47,360 SO WE THEN DID EXACTLY THE SAME 1428 00:51:47,360 --> 00:51:49,080 THING THAT WIDE DONE FOR SINGLE 1429 00:51:49,080 --> 00:51:53,640 CELL IN BREAST CANCER, WE 1430 00:51:53,640 --> 00:51:54,720 ASKED -- TO INVERT THE SIGNATURE 1431 00:51:54,720 --> 00:51:55,240 OF THE T REGS. 1432 00:51:55,240 --> 00:51:58,800 SO WE STARTED WITH ABOUT 1500 1433 00:51:58,800 --> 00:52:00,360 DRUGS FDA-APPROVED, SELECTED THE 1434 00:52:00,360 --> 00:52:02,600 ONES THAT -- DID THOSE RESPONSE 1435 00:52:02,600 --> 00:52:04,080 CURVES FOR THOSE AND WE SELECTED 1436 00:52:04,080 --> 00:52:07,800 THE TOP 86, AND THESE WERE NOW 1437 00:52:07,800 --> 00:52:12,600 TO PERTURB INFILTRATING T REGS, 1438 00:52:12,600 --> 00:52:16,400 AND THEN WE GENERATED THE RNA 1439 00:52:16,400 --> 00:52:18,560 SEQ PROFILES AND -- WHAT YOU SEE 1440 00:52:18,560 --> 00:52:24,600 HERE THE THREE DRUGS, WE WERE 1441 00:52:24,600 --> 00:52:26,360 ESSENTIALLY ABLE TO INVERT 1442 00:52:26,360 --> 00:52:27,480 INFILTRATING VERSUS -- IN 1443 00:52:27,480 --> 00:52:28,720 VIRTUALLY EACH ONE OF THE 32 1444 00:52:28,720 --> 00:52:30,840 PATIENTS THAT WE HAD COLLECTED 1445 00:52:30,840 --> 00:52:32,800 EXCEPT FOR MAYBE ONE OR TWO, BUT 1446 00:52:32,800 --> 00:52:43,000 WHAT WAS EVEN MORE STRIKING IS 1447 00:52:43,000 --> 00:52:49,240 GENERAL GEM SITE BEAN IS THE 1448 00:52:49,240 --> 00:52:50,840 ONLY ONE YOU CAN REALLY VALIDATE 1449 00:52:50,840 --> 00:53:00,520 BECAUSE IT HAS A GRAD RULE 1450 00:53:00,520 --> 00:53:03,920 RESPONSE, -- IN A WAY THAT THEY 1451 00:53:03,920 --> 00:53:06,640 KEL T REGS BUT NOT THE 1452 00:53:06,640 --> 00:53:08,720 NON-INFILTRATING T REGS FOR THE 1453 00:53:08,720 --> 00:53:10,400 CELLS THAT ARE INVOLVED IN 1454 00:53:10,400 --> 00:53:13,000 IMMUNE RESPONSE. 1455 00:53:13,000 --> 00:53:14,400 SO THIS SHOWS THE RESPONSE OF 1456 00:53:14,400 --> 00:53:16,400 TUMORS TREATED IN VIVO WITH 1457 00:53:16,400 --> 00:53:17,200 12 MILLIGRAMS PER KILOGRAM WHICH 1458 00:53:17,200 --> 00:53:19,040 IS ABOUT 15 FOLD LOWER THAN WHAT 1459 00:53:19,040 --> 00:53:21,240 YOU WOULD NORMALLY USE FOR 1460 00:53:21,240 --> 00:53:25,320 CLINICAL STUDY, WHICH IS AROUND 1461 00:53:25,320 --> 00:53:25,680 150 MILLIGRAMS. 1462 00:53:25,680 --> 00:53:27,600 YOU CAN SEE THERE'S ABSOLUTELY 1463 00:53:27,600 --> 00:53:33,760 NO EFFECT IN A COMPROMISED -- 1464 00:53:33,760 --> 00:53:40,120 STRONG EFFECT -- THAT BY ITSELF 1465 00:53:40,120 --> 00:53:41,160 HATION 1466 00:53:41,160 --> 00:53:43,200 THAT HAS ACTUALLY NO RESPONSE BY 1467 00:53:43,200 --> 00:53:43,480 ITSELF. 1468 00:53:43,480 --> 00:53:46,520 IF YOU GO TO 120 MG, YOU NOW SEE 1469 00:53:46,520 --> 00:53:48,240 THE NSG MODEL STARTING TO 1470 00:53:48,240 --> 00:53:50,640 RESPOND A LITTLE BIT, BUT YOU 1471 00:53:50,640 --> 00:53:52,800 STILL PRESERVE THE RESPONSE IN 1472 00:53:52,800 --> 00:53:54,200 THE IMMUNOCOMPETENT MODEL. 1473 00:53:54,200 --> 00:53:56,240 SO I'LL STOP HERE, JUST WANTED 1474 00:53:56,240 --> 00:53:57,440 TO SAY THAT FOR THE OTHER PEOPLE 1475 00:53:57,440 --> 00:53:58,920 THAT ARE ON THIS CALL THAT ARE 1476 00:53:58,920 --> 00:54:01,200 NOT INTERESTED IN CANCER, THIS 1477 00:54:01,200 --> 00:54:02,640 IS NOT THE WAY CANCER CELLS 1478 00:54:02,640 --> 00:54:04,080 WORK, THIS IS THE WAY ALL CELLS 1479 00:54:04,080 --> 00:54:06,200 WORK, AND SO WE HAVE USED THIS 1480 00:54:06,200 --> 00:54:08,320 MODEL TO ESSENTIALLY DO STUDIES 1481 00:54:08,320 --> 00:54:12,040 IN BIOLOGY, MANY DEGENERATIVE 1482 00:54:12,040 --> 00:54:13,440 DISEASES, CARDIOVASCULAR 1483 00:54:13,440 --> 00:54:14,560 DISEASE, JUST PUBLISHED WITH THE 1484 00:54:14,560 --> 00:54:16,400 RILEY GROUP, TYPE 2 DIABETES 1485 00:54:16,400 --> 00:54:18,280 JUST CAME OUT, AND THEN 1486 00:54:18,280 --> 00:54:19,200 SOMETHING THAT WE'RE VERY 1487 00:54:19,200 --> 00:54:21,000 EXCITED ABOUT IS THE FACT THAT 1488 00:54:21,000 --> 00:54:23,080 NOW THAT WE KNOW HOW TO DO THESE 1489 00:54:23,080 --> 00:54:24,760 ANALYSES, WE CAN LITERALLY TAKE 1490 00:54:24,760 --> 00:54:28,200 ANY CELL STATE THAT YOU START 1491 00:54:28,200 --> 00:54:29,560 FROM AND ANY CELL STATE YOU WANT 1492 00:54:29,560 --> 00:54:32,120 TO END IN AND DEVISE EITHER 1493 00:54:32,120 --> 00:54:33,360 GENETIC OR PHARMACOLOGICAL 1494 00:54:33,360 --> 00:54:34,440 RECIPE TO MAKE IT HAPPEN. 1495 00:54:34,440 --> 00:54:37,720 SO IF WE SAY THIS IS AN EXAMPLE 1496 00:54:37,720 --> 00:54:41,480 WHERE WE IDENTIFIED -- AS A 1497 00:54:41,480 --> 00:54:42,320 SINGLE TRANSCRIPTION FACTOR THAT 1498 00:54:42,320 --> 00:54:45,400 CAN BE PROGRAMMED -- HUMAN 1499 00:54:45,400 --> 00:54:48,040 LINEAGE PROGENITORS -- HSC STATE 1500 00:54:48,040 --> 00:54:49,160 THAT WAS JUST PUBLISHED. 1501 00:54:49,160 --> 00:54:50,680 SO JUST WANTED TO THANK A LOT OF 1502 00:54:50,680 --> 00:54:54,200 PEOPLE AND ALSO THE NIH FOR VERY 1503 00:54:54,200 --> 00:54:56,320 GENEROUS SUPPORT TO THIS 1504 00:54:56,320 --> 00:54:56,600 RESEARCH. 1505 00:54:56,600 --> 00:54:58,200 THIS IS NOT HAPPENING IN ONE 1506 00:54:58,200 --> 00:55:01,840 LAB, IT'S A HUMONGOUS NUMBER OF 1507 00:55:01,840 --> 00:55:03,040 COLLABORATORS THAT CONTRIBUTED 1508 00:55:03,040 --> 00:55:04,360 TO THESE AND FOUNDATIONS. 1509 00:55:04,360 --> 00:55:05,200 ALSO EVERYTHING WE DO IS MADE 1510 00:55:05,200 --> 00:55:08,240 AVAILABLE TO THE COMMUNITY,, 1511 00:55:08,240 --> 00:55:12,080 BOTH OUR -- THAT WE USE ARE ALL 1512 00:55:12,080 --> 00:55:14,200 AVAILABLE TO ADGENE, THE 1513 00:55:14,200 --> 00:55:16,800 SOFTWARE WE USE, OR THROUGH THE 1514 00:55:16,800 --> 00:55:19,640 WEBSITES IN OUR SHARED RESOURCES 1515 00:55:19,640 --> 00:55:20,000 OF THE DATA. 1516 00:55:20,000 --> 00:55:26,400 THANK YOU. 1517 00:55:26,400 --> 00:55:28,280 >> FANTASTIC. 1518 00:55:28,280 --> 00:55:31,080 THANK YOU VERY MUCH, ANDREA. 1519 00:55:31,080 --> 00:55:32,400 AS USUAL, I'VE GOT A LOT OF 1520 00:55:32,400 --> 00:55:32,760 QUESTIONS. 1521 00:55:32,760 --> 00:55:40,920 LET'S SEE IF I CAN STUMP YOU. 1522 00:55:40,920 --> 00:55:42,480 BUT I'LL JUST GET START AND THEN 1523 00:55:42,480 --> 00:55:46,880 PLEASE, ANYBODY IN THE AUDIENCE 1524 00:55:46,880 --> 00:55:48,880 WHO HAS A QUESTION, DO TYPE IN 1525 00:55:48,880 --> 00:55:50,280 YOUR QUESTIONS IN THE CHAT AND 1526 00:55:50,280 --> 00:55:55,000 THEN WE WILL -- THEN I'LL REPEAT 1527 00:55:55,000 --> 00:56:00,240 THEM. 1528 00:56:00,240 --> 00:56:00,840 WHERE TO START. 1529 00:56:00,840 --> 00:56:02,280 SO I'VE BEEN CONVINCED FOR A 1530 00:56:02,280 --> 00:56:07,160 LONG TIME THAT YOU CAN FIND 1531 00:56:07,160 --> 00:56:08,600 THESE MASTER REGULATORS, SO AS 1532 00:56:08,600 --> 00:56:15,400 YOU KNOW, WE TAKE KIND OF A MUCH 1533 00:56:15,400 --> 00:56:17,040 MORE TRADITIONAL, I WOULD SAY, 1534 00:56:17,040 --> 00:56:18,360 APPROACH TO FINDING AND 1535 00:56:18,360 --> 00:56:21,320 DEVELOPING NEW THERAPIES. 1536 00:56:21,320 --> 00:56:23,000 WHICH ALSO WORKS, I WOULD SAY. 1537 00:56:23,000 --> 00:56:23,400 SO -- 1538 00:56:23,400 --> 00:56:24,400 >> IT WORKS REALLY WELL. 1539 00:56:24,400 --> 00:56:28,760 >> SO THE QUESTION IS, SO WHAT 1540 00:56:28,760 --> 00:56:30,520 WE'VE FOUND, GOING WITH MY FIRST 1541 00:56:30,520 --> 00:56:31,760 QUESTION, WHAT WE'VE FOUND IS OF 1542 00:56:31,760 --> 00:56:33,240 COURSE WHAT EVERYBODY HAS FOUND 1543 00:56:33,240 --> 00:56:35,760 IN CANCER, IS THAT SINGLE AGENT 1544 00:56:35,760 --> 00:56:36,840 TREATMENT OF ANYTHING, I DON'T 1545 00:56:36,840 --> 00:56:38,800 CARE WHAT, IS INSUFFICIENT. 1546 00:56:38,800 --> 00:56:40,640 OKAY? 1547 00:56:40,640 --> 00:56:45,280 IT'S VERY -- THERE ARE 1548 00:56:45,280 --> 00:56:46,440 EXCEPTIONS BCR-ABLE, BUT THEY'RE 1549 00:56:46,440 --> 00:56:48,320 RARE. 1550 00:56:48,320 --> 00:56:53,960 SO WE HAVE GONE TO DEVELOPING 1551 00:56:53,960 --> 00:56:55,360 FAIRLY COMPLEX MIXTURES OF 1552 00:56:55,360 --> 00:56:55,920 TARGETED AGENTS. 1553 00:56:55,920 --> 00:56:59,400 SO MY QUESTION IS, 1554 00:56:59,400 --> 00:57:00,400 COMPUTATIONALLY, HOW DOES THAT 1555 00:57:00,400 --> 00:57:03,600 WORK OUT FOR YOU? 1556 00:57:03,600 --> 00:57:06,200 CAN YOU -- SO YOU DO -- THERE'S 1557 00:57:06,200 --> 00:57:07,000 TWO DIFFERENT WAYS OF THINKING 1558 00:57:07,000 --> 00:57:09,360 ABOUT IT, YOU HAVE TWO PARALLEL 1559 00:57:09,360 --> 00:57:11,920 PATHWAYS THAT CONTROL DIRN 1560 00:57:11,920 --> 00:57:13,120 THINGS, YOU WANT TO TARGET EACH 1561 00:57:13,120 --> 00:57:14,840 OR YOU WANT TO TARGET ONE VERY 1562 00:57:14,840 --> 00:57:20,960 IMPORTANT PATHWAY TWICE, SO HOW 1563 00:57:20,960 --> 00:57:22,400 DOES YOUR MATHEMATICS SHOW YOU 1564 00:57:22,400 --> 00:57:23,200 THE WAY FORWARD? 1565 00:57:23,200 --> 00:57:24,920 WHICH IS THE BEST WAY TO COMBINE 1566 00:57:24,920 --> 00:57:27,400 DRUGS IN MULTI-TARGETED THERAPY? 1567 00:57:27,400 --> 00:57:29,120 >> SO LET ME FIRST ADDRESS THE 1568 00:57:29,120 --> 00:57:30,240 QUESTION ITSELF BECAUSE THERE'S 1569 00:57:30,240 --> 00:57:33,200 A LITTLE BIT OF PHILOSOPHY 1570 00:57:33,200 --> 00:57:34,480 BEHIND IT, NEEDS TO BE BROUGHT 1571 00:57:34,480 --> 00:57:35,160 TO BEAR. 1572 00:57:35,160 --> 00:57:36,920 THERE ARE REALLY THREE 1573 00:57:36,920 --> 00:57:38,840 FUNDAMENTAL REASONS WHY SINGLE 1574 00:57:38,840 --> 00:57:39,720 AGENT THERAPY FAILS. 1575 00:57:39,720 --> 00:57:41,080 ONE REASON IS THAT DRUGS, MOST 1576 00:57:41,080 --> 00:57:43,120 DRUGS HAVE ACTUALLY BEEN BUILT 1577 00:57:43,120 --> 00:57:44,200 TO TARGET INDIVIDUAL PROTEINS 1578 00:57:44,200 --> 00:57:47,680 THAT ARE ACTUALLY ONCOGENES. 1579 00:57:47,680 --> 00:57:50,200 OR THAT ARE INVOLVED IN AN 1580 00:57:50,200 --> 00:57:50,960 ONCOGENIC PATHWAY. 1581 00:57:50,960 --> 00:57:51,920 SO YOU'RE STARTING WITH DRUGS 1582 00:57:51,920 --> 00:57:54,560 THAT ARE ALMOST BUILT TO FAIL 1583 00:57:54,560 --> 00:57:59,240 BECAUSE WITHIN THE CELLS, WE 1584 00:57:59,240 --> 00:58:00,760 NEED THE MAP, YOU HAVE MORE 1585 00:58:00,760 --> 00:58:02,600 MUTATIONAL PATTERNS THAT CAN 1586 00:58:02,600 --> 00:58:04,280 INDUCE A TUMOR THAN ATOMS IN THE 1587 00:58:04,280 --> 00:58:06,800 UNIVERSE. 1588 00:58:06,800 --> 00:58:11,000 THERE'S ABOUT -- FROM THE 1589 00:58:11,000 --> 00:58:13,960 ONCOGENE WE ALREADY KNOW ABOUT 1590 00:58:13,960 --> 00:58:16,000 AND THERE'S 10 TO THE 80 ATOMS 1591 00:58:16,000 --> 00:58:16,720 IN THE UNIVERSE. 1592 00:58:16,720 --> 00:58:19,080 SO EVERY SINGLE CELL YOU HAVE IN 1593 00:58:19,080 --> 00:58:21,200 THE TUMOR WILL ACTUALLY ACCRUE 1594 00:58:21,200 --> 00:58:23,000 ADDITIONAL MUTATIONS THAT WILL 1595 00:58:23,000 --> 00:58:24,400 DEFEAT SINGLE CELL THERAPY 1596 00:58:24,400 --> 00:58:27,280 DIRECTED AT AN ONCOGENE. 1597 00:58:27,280 --> 00:58:28,320 THAT'S NUMBER ONE. 1598 00:58:28,320 --> 00:58:29,000 NUMBER TWO IS THAT YOU HAVE 1599 00:58:29,000 --> 00:58:31,280 WITHIN THE TUMOR A DIFFERENT 1600 00:58:31,280 --> 00:58:32,240 SUBPOPULATION THAT HAVE 1601 00:58:32,240 --> 00:58:33,440 COMPLETELY DIFFERENT DRUG 1602 00:58:33,440 --> 00:58:36,120 SENSITIVITY. 1603 00:58:36,120 --> 00:58:39,360 THIS IS ACTUALLY EPIGENETICALLY 1604 00:58:39,360 --> 00:58:39,640 DETERMINED. 1605 00:58:39,640 --> 00:58:43,720 AS I SHOWED YOU YOU CAN 1606 00:58:43,720 --> 00:58:44,840 LITERALLY REPROGRAM THE CELLS 1607 00:58:44,840 --> 00:58:46,320 WITHOUT HAVING TO CHANGE THEIR 1608 00:58:46,320 --> 00:58:46,800 GENETICS. 1609 00:58:46,800 --> 00:58:48,360 AND THE THIRD ONE IS THAT CELLS 1610 00:58:48,360 --> 00:58:50,840 ARE INCREDIBLY GOOD AT 1611 00:58:50,840 --> 00:58:51,840 PERFORMING WHAT'S CALLED 1612 00:58:51,840 --> 00:58:53,800 ADAPTIVE RESPONSE, SO THEY 1613 00:58:53,800 --> 00:58:58,400 REALLY REWIRE THEIR NETWORKS IN 1614 00:58:58,400 --> 00:59:01,280 RESPONSE TO THE DRUG, THIS 1615 00:59:01,280 --> 00:59:04,200 BEAUTIFUL STUDY THAT SHOWS THE 1616 00:59:04,200 --> 00:59:06,040 MAP KINASE PATHWAYS -- 1617 00:59:06,040 --> 00:59:08,360 INHIBITORS, SO TAKEN TOGETHER, 1618 00:59:08,360 --> 00:59:09,720 THOSE THREE REALLY FORMULATE A 1619 00:59:09,720 --> 00:59:11,200 MANIFESTO FAILURE OF SINGLE 1620 00:59:11,200 --> 00:59:13,040 AGENTS, RIGHT? 1621 00:59:13,040 --> 00:59:15,160 SO WHAT WE HAVE OBSERVED IS THAT 1622 00:59:15,160 --> 00:59:17,200 WHEN YOU GO AND PERTURB THE 1623 00:59:17,200 --> 00:59:18,480 CELLS WITH DRUGS, YOU ACTUALLY 1624 00:59:18,480 --> 00:59:21,280 DON'T CHANGE THE POPULATION. 1625 00:59:21,280 --> 00:59:23,480 THEIR A STABLE POPULATION, YOU 1626 00:59:23,480 --> 00:59:24,640 ADD AT THE BEGINNING BEFORE 1627 00:59:24,640 --> 00:59:26,440 TREATMENT ARE ALMOST NEVER 1628 00:59:26,440 --> 00:59:26,680 CHANGED. 1629 00:59:26,680 --> 00:59:28,440 IN SOME CASES YOU DO, IN 1630 00:59:28,440 --> 00:59:31,360 PROSTATE CANCER, YOU GET A 1631 00:59:31,360 --> 00:59:33,240 COMPLETELY NEW POPULATION OF 1632 00:59:33,240 --> 00:59:33,880 NEUROENDOCRINE TUMOR CELLS WHEN 1633 00:59:33,880 --> 00:59:36,000 YOU TREAT THE OTHER WITH -- AND 1634 00:59:36,000 --> 00:59:38,080 YOU HAVE A -- MUTATION BUT IT'S 1635 00:59:38,080 --> 00:59:40,280 VERY RARE. 1636 00:59:40,280 --> 00:59:42,600 IN FACT I SHOWED YOU EVEN FOR 1637 00:59:42,600 --> 00:59:43,480 THE BREAST CANCER -- COMPLETED 1638 00:59:43,480 --> 00:59:44,760 FOR DRUGS THE STATES REMAIN THE 1639 00:59:44,760 --> 00:59:47,800 SAME. 1640 00:59:47,800 --> 00:59:49,400 SO WHAT WE DO IS INSTEAD OF 1641 00:59:49,400 --> 00:59:50,960 FOCUSING ON THE PATHWAY OR THE 1642 00:59:50,960 --> 00:59:52,560 ONCOPROTEINS, WE FOCUS ON THE 1643 00:59:52,560 --> 00:59:53,840 STATE OF THE CELL. 1644 00:59:53,840 --> 00:59:54,880 ESSENTIALLY ASK WHAT ARE THE 1645 00:59:54,880 --> 00:59:56,720 PROTEINS THAT MAINTAIN THE STATE 1646 00:59:56,720 --> 01:00:03,720 AND THOUSAND CAN WE HOW CAN WE TARGET THE M 1647 01:00:03,720 --> 01:00:04,120 PHARMACOLOGICALLY. 1648 01:00:04,120 --> 01:00:05,240 MANY OF THE PROTEINS YOU TRY TO 1649 01:00:05,240 --> 01:00:06,880 TARGET ARE IN FACT THE UPSTREAM 1650 01:00:06,880 --> 01:00:08,120 MODULATORS OF THE MASTER 1651 01:00:08,120 --> 01:00:09,160 REGULATORS IN THE CELL. 1652 01:00:09,160 --> 01:00:11,680 SO IF YOU REMEMBER FROM THAT 1653 01:00:11,680 --> 01:00:14,360 PAPER ON -- WITH RICARDO, WE DID 1654 01:00:14,360 --> 01:00:15,840 THIS ANALYSES IN A KIND OF POOR 1655 01:00:15,840 --> 01:00:18,360 MANUWAI AND WE SHOW THAT -- KB 1656 01:00:18,360 --> 01:00:19,560 WAS THE REPORTER FOR THE 1657 01:00:19,560 --> 01:00:21,200 ACTIVITY OF ALL THESE MUTATIONS 1658 01:00:21,200 --> 01:00:22,680 IN ITS SUBSTREAM PATHWAYS. 1659 01:00:22,680 --> 01:00:25,000 SO WHEN YOU FIND A DRUG THAT 1660 01:00:25,000 --> 01:00:26,800 ACTS ON -- TK YOU FIND A DRUG 1661 01:00:26,800 --> 01:00:28,600 THAT IS ACTUALLY DISABLING THE 1662 01:00:28,600 --> 01:00:30,000 MASTER REGULATORS THROUGH A 1663 01:00:30,000 --> 01:00:31,120 MECHANISM THAT IS COMPLEX 1664 01:00:31,120 --> 01:00:32,040 BECAUSE THERE'S A FEW STEPS TO 1665 01:00:32,040 --> 01:00:36,600 GET THERE. 1666 01:00:36,600 --> 01:00:39,400 NOW WE'RE DOING THIS IN 1667 01:00:39,400 --> 01:00:40,920 COLLABORATION WITH A LOT OF 1668 01:00:40,920 --> 01:00:42,040 PHARMACEUTICAL COMPANIES, 1669 01:00:42,040 --> 01:00:43,080 INSTEAD OF TARGETING THESE GENES 1670 01:00:43,080 --> 01:00:45,160 THAT ARE UPSTREAM ARE ACTUALLY 1671 01:00:45,160 --> 01:00:47,240 TARGETING DIRECTLY FROM -- TO -- 1672 01:00:47,240 --> 01:00:49,600 TECHNOLOGY ARE TARGETING 1673 01:00:49,600 --> 01:00:50,720 DIRECTLY THE MASTER REGULATOR. 1674 01:00:50,720 --> 01:00:54,080 >> THAT MAKES SENSE FOR THE 1675 01:00:54,080 --> 01:00:54,400 FUTURE. 1676 01:00:54,400 --> 01:00:57,560 FOR SURE. 1677 01:00:57,560 --> 01:01:01,720 >> IN COMBINATION -- DRUGS THAT 1678 01:01:01,720 --> 01:01:02,440 TARGET DIFFERENT POPULATION AND 1679 01:01:02,440 --> 01:01:05,120 YOU COMBINE THEM TOGETHER, YOU 1680 01:01:05,120 --> 01:01:06,320 FIND THINGS THAT ARE 1681 01:01:06,320 --> 01:01:06,600 SYNERGISTIC. 1682 01:01:06,600 --> 01:01:09,640 >> SO FOR EXAMPLE, LET'S TAKE 1683 01:01:09,640 --> 01:01:14,120 SOMETHING I'M VERY FAMILIAR 1684 01:01:14,120 --> 01:01:14,600 WITH. 1685 01:01:14,600 --> 01:01:15,480 YOU HAVE A LOT OF THINGS THAT 1686 01:01:15,480 --> 01:01:16,920 DON'T WORK AS A SINGLE AGENT, 1687 01:01:16,920 --> 01:01:19,360 BUT IF YOU SIMPLY INHIBIT 1688 01:01:19,360 --> 01:01:21,480 BCL2 IN A LYMPHOMA, THEN THAT'S 1689 01:01:21,480 --> 01:01:23,120 THE BALL GAME. 1690 01:01:23,120 --> 01:01:25,120 THAT BASICALLY -- AND SO I DON'T 1691 01:01:25,120 --> 01:01:27,640 KNOW WHETHER YOU WOULD REFER TO 1692 01:01:27,640 --> 01:01:31,200 BCL2 AS A MASTER REGULATOR. 1693 01:01:31,200 --> 01:01:32,400 IT IS THIS MASTER THING IN THE 1694 01:01:32,400 --> 01:01:32,720 CELL. 1695 01:01:32,720 --> 01:01:36,240 SO HOW DOES THAT FIT IN YOUR 1696 01:01:36,240 --> 01:01:36,560 PHILOSOPHY? 1697 01:01:36,560 --> 01:01:39,000 >> THE PROBLEM WITH BCL2 AND 1698 01:01:39,000 --> 01:01:40,400 MASTER REGULATORS, A LONG TIME 1699 01:01:40,400 --> 01:01:42,640 AGO, I THINK IT WAS IN 2010, SO 1700 01:01:42,640 --> 01:01:45,040 IT DEFINITELY COMES UP AS MASTER 1701 01:01:45,040 --> 01:01:46,600 REGULATOR PROTEINS, BUT -- 1702 01:01:46,600 --> 01:01:49,000 >> SORRY, IT'S NOT A MASTER 1703 01:01:49,000 --> 01:01:52,680 REGULATOR IN TERMS OF GENE 1704 01:01:52,680 --> 01:01:53,160 EXPRESSION. 1705 01:01:53,160 --> 01:01:54,600 >> IT'S NOT A DIRECT REGULATOR 1706 01:01:54,600 --> 01:01:57,160 BUT WHAT HAPPENS IS THAT 1707 01:01:57,160 --> 01:01:58,200 SUBSIGNALING PROTEINS ARE SO 1708 01:01:58,200 --> 01:01:59,600 CLOSE TO THE TRANSCRIPTIONAL 1709 01:01:59,600 --> 01:02:04,400 LAYER THAT ESSENTIALLY ACT AS -- 1710 01:02:04,400 --> 01:02:05,880 TRANSCRIPTIONAL REGULATORS 1711 01:02:05,880 --> 01:02:06,960 BECAUSE THEY DIRECTLY 1712 01:02:06,960 --> 01:02:07,960 PHOSPHORYLATE OR -- A 1713 01:02:07,960 --> 01:02:09,440 TRANSCRIPTION FACTOR, SO YOU CAN 1714 01:02:09,440 --> 01:02:10,600 REALLY TRANSFER THEIR ACTIVITY 1715 01:02:10,600 --> 01:02:12,280 ALMOST AS IF IT WERE A 1716 01:02:12,280 --> 01:02:13,400 TRANSCRIPTION FACTOR. 1717 01:02:13,400 --> 01:02:14,840 SO THE ANALYSIS IS NOT AS GOOD 1718 01:02:14,840 --> 01:02:17,720 FOR THOSE PROTEINS AS IT IS FOR 1719 01:02:17,720 --> 01:02:20,000 TRANSCRIPTION FACTOR, 80% TO 1720 01:02:20,000 --> 01:02:25,120 ABOUT 65 PERCIVAL DAITION RATE 65% -- SO I T STILL DOES 1721 01:02:25,120 --> 01:02:28,400 REALLY WELL. 1722 01:02:28,400 --> 01:02:30,040 -- GLUCOCORTICOID 1723 01:02:30,040 --> 01:02:31,840 SENSITIVITY -- IS NOT A 1724 01:02:31,840 --> 01:02:32,400 TRANSCRIPTIONAL REGULATORS. 1725 01:02:32,400 --> 01:02:33,640 BUT IN GENERAL, WE WOULD SAY 1726 01:02:33,640 --> 01:02:36,360 THAT PROBABLY BCL2 IS 1727 01:02:36,360 --> 01:02:39,160 IMPLEMENTING AN EVEN MORE 1728 01:02:39,160 --> 01:02:40,920 TRANSCRIPTIONAL SPECIFIC CONTROL 1729 01:02:40,920 --> 01:02:44,200 MODULE THAT BCL2 IN SOME WAY 1730 01:02:44,200 --> 01:02:45,800 INHIBITS IN AN AMAZINGLY GOOD 1731 01:02:45,800 --> 01:02:51,880 WAY SO YOU ARE -- BY 1732 01:02:51,880 --> 01:02:52,440 ACTIVATING -- 1733 01:02:52,440 --> 01:02:53,120 >> SO I HAVE ONE QUESTION FROM 1734 01:02:53,120 --> 01:02:58,480 THE AUDIENCE THAT GIBES WITH ONE 1735 01:02:58,480 --> 01:03:00,200 I WAS GOING TO ASK. 1736 01:03:00,200 --> 01:03:03,160 THE QUESTION WAS FROM -- YEN, 1737 01:03:03,160 --> 01:03:05,040 SORRY FOR BUTCHERING YOUR NAME, 1738 01:03:05,040 --> 01:03:09,440 AND IT SAYS MANY CANCER DRUG 1739 01:03:09,440 --> 01:03:11,440 COMPANION DIAGNOSTICS ARE 1740 01:03:11,440 --> 01:03:15,480 MUTATION-BASED. 1741 01:03:15,480 --> 01:03:17,520 HOW ARE THE MUTATION INFORMATION 1742 01:03:17,520 --> 01:03:18,760 USED IN COMBINATION WITH THE 1743 01:03:18,760 --> 01:03:19,680 MASTER REGULATORS? 1744 01:03:19,680 --> 01:03:21,600 I'M GOING TO BROADEN THIS OUT IN 1745 01:03:21,600 --> 01:03:23,680 GENERAL TO SAY IF YOU ARE GOING 1746 01:03:23,680 --> 01:03:29,000 FOR REGULATORY APPROVAL FOR A 1747 01:03:29,000 --> 01:03:32,360 DRUG THAT'S TARGETING MASTER 1748 01:03:32,360 --> 01:03:33,400 REGULATORS OR COMBINATION, WHAT 1749 01:03:33,400 --> 01:03:36,640 WOULD IT LOOK LIKE, WHAT WOULD 1750 01:03:36,640 --> 01:03:39,640 BE YOUR DIAGNOSTIC THAT WOULD 1751 01:03:39,640 --> 01:03:43,760 LEAD TO IDENTIFYING THE RESPONSE 1752 01:03:43,760 --> 01:03:44,600 OF PATIENTS? 1753 01:03:44,600 --> 01:03:46,200 AND HOW DOES THAT FIT WITH WHAT 1754 01:03:46,200 --> 01:03:50,680 HE OR SHE, I'M SORRY, ASKS ABOUT 1755 01:03:50,680 --> 01:03:51,800 MUTATIONS AS PART OF THE 1756 01:03:51,800 --> 01:03:52,600 DIAGNOSIS? 1757 01:03:52,600 --> 01:03:54,960 >> SO WE NOW HAVE, AS I TOLD 1758 01:03:54,960 --> 01:03:56,800 YOU, A CLIA CERTIFIED ASSAY THAT 1759 01:03:56,800 --> 01:03:59,080 CAN BE USED TO USE THE ACTIVITY 1760 01:03:59,080 --> 01:04:02,120 OF ANY PROTEIN AS A REPORTER FOR 1761 01:04:02,120 --> 01:04:03,000 THE ACTIVITY OF A DRUG. 1762 01:04:03,000 --> 01:04:05,000 SO WE HAVE ALREADY PUBLISHED 1763 01:04:05,000 --> 01:04:06,880 SEVERAL MANUSCRIPTS WHERE WE 1764 01:04:06,880 --> 01:04:10,320 ACTUALLY PRODUCED PREDICTORS OF 1765 01:04:10,320 --> 01:04:11,680 ACTIVITY OF DRUGS IN PATIENTS, 1766 01:04:11,680 --> 01:04:16,280 THIS IS NOT IN MY -- CELL LINES, 1767 01:04:16,280 --> 01:04:23,240 INCLUDING -- INCLUDING -- IN 1768 01:04:23,240 --> 01:04:25,120 AML -- AND GLIOMA, WE'RE NOW 1769 01:04:25,120 --> 01:04:26,640 ACTUALLY STARTING, JUST GOT 1770 01:04:26,640 --> 01:04:28,200 FUNDED TO START A STUDY IN 1771 01:04:28,200 --> 01:04:32,280 GLIOMA BASED ON THAT BIOMARKER, 1772 01:04:32,280 --> 01:04:34,480 AND SO -- LAB HAS PUBLISHED A 1773 01:04:34,480 --> 01:04:35,640 PAPER WHERE IT SHOWS THAT AMONG 1774 01:04:35,640 --> 01:04:38,480 ALL THE THINGS YOU CAN USE IN 1775 01:04:38,480 --> 01:04:40,760 THE -- LANDSCAPE TO PREDICT DRUG 1776 01:04:40,760 --> 01:04:42,160 ACTIVITY, GENETICS IS ABSOLUTELY 1777 01:04:42,160 --> 01:04:43,040 ON THE BOTTOM. 1778 01:04:43,040 --> 01:04:44,720 I FOUND IT VERY REFRESHING TO 1779 01:04:44,720 --> 01:04:47,760 SEE THAT THAT RESULTS -- SORT OF 1780 01:04:47,760 --> 01:04:49,120 PROPOSING FOR A LONG TIME CAME 1781 01:04:49,120 --> 01:04:52,440 FROM A LAB THAT IS AT THE BROAD 1782 01:04:52,440 --> 01:04:54,400 BUT BASICALLY REALITY. 1783 01:04:54,400 --> 01:04:56,000 THE -- MOST INFORMATIVE ONE IN 1784 01:04:56,000 --> 01:04:57,800 HIS ANALYSIS FOR GENE 1785 01:04:57,800 --> 01:04:58,400 EXPRESSION, WHAT WAS SHOWN IS 1786 01:04:58,400 --> 01:05:01,800 THAT THE PROTEIN ACTIVITY IS 1787 01:05:01,800 --> 01:05:05,160 DRAMATICALLY MORE EFFECTIVE THAN 1788 01:05:05,160 --> 01:05:05,800 GENE EXPRESSION IMPACT. 1789 01:05:05,800 --> 01:05:07,240 WHAT HAPPENS WITH GENE 1790 01:05:07,240 --> 01:05:09,800 EXPRESSION, WHEN YOU BUILD A 1791 01:05:09,800 --> 01:05:10,760 CLASSIFIER, THE MORE GENES YOU 1792 01:05:10,760 --> 01:05:12,280 USE, THE BETTER THE 1793 01:05:12,280 --> 01:05:13,600 CLASSIFICATION BECOMES. 1794 01:05:13,600 --> 01:05:15,680 THAT'S A CLASSICAL SIGN OF 1795 01:05:15,680 --> 01:05:16,560 OVERTRAINING, OVERFITTING. 1796 01:05:16,560 --> 01:05:18,000 WITH PROTEIN ACTIVITY, YOU 1797 01:05:18,000 --> 01:05:21,040 BASICALLY GET BETTER FOR THE 1798 01:05:21,040 --> 01:05:22,840 FIRST SIX OR SEVEN PROTEINS, 1799 01:05:22,840 --> 01:05:23,960 SOMETIMES THE FIRST FOUR, AND IT 1800 01:05:23,960 --> 01:05:25,560 ACTUALLY GETS WORSE OR DOESN'T 1801 01:05:25,560 --> 01:05:25,920 IMPROVE ANYMORE. 1802 01:05:25,920 --> 01:05:29,120 SO BASICALLY THIS TELLS YOU THAT 1803 01:05:29,120 --> 01:05:30,360 THAT'S THE MINIMUM NUMBER OF 1804 01:05:30,360 --> 01:05:32,040 MARKERS YOU CAN USE AND IT'S 1805 01:05:32,040 --> 01:05:33,600 VERY GOOD BECAUSE CLINICALLY, 1806 01:05:33,600 --> 01:05:35,000 USING ONLY FOUR PROTEINS IS A 1807 01:05:35,000 --> 01:05:37,360 GREAT WAY TO DO IT EVEN IN SITU 1808 01:05:37,360 --> 01:05:39,000 INSTEAD OF HAVING TO DO GENE 1809 01:05:39,000 --> 01:05:40,480 EXPRESSION AND LOOKING AT A 1810 01:05:40,480 --> 01:05:43,800 PANEL WITH -- SO THIS IS ALL 1811 01:05:43,800 --> 01:05:44,760 PUBLISHED, I CAN SEND YOU THE 1812 01:05:44,760 --> 01:05:45,000 REFERENCE. 1813 01:05:45,000 --> 01:05:47,640 >> NO, NO, NO, THAT'S FINE. 1814 01:05:47,640 --> 01:05:49,480 I'M GOING TO NOT LET YOU GET THE 1815 01:05:49,480 --> 01:05:55,440 LAST WORD ON THAT SUBJECT. 1816 01:05:55,440 --> 01:05:57,040 SO WHAT CLEARLY INDIVIDUAL GENE 1817 01:05:57,040 --> 01:06:00,920 MUTATIONS HAVE THEIR LIMITATIONS 1818 01:06:00,920 --> 01:06:02,200 IN -- BUT WHAT WE'VE BEEN 1819 01:06:02,200 --> 01:06:03,400 FOCUSED ON AS YOU PROBABLY KNOW, 1820 01:06:03,400 --> 01:06:05,480 AND IT'S RELATED, I THINK, TO 1821 01:06:05,480 --> 01:06:10,000 THE MASTER REGULATOR IDEA, IS JE 1822 01:06:10,000 --> 01:06:10,400 GENETIC SUBTYPES. 1823 01:06:10,400 --> 01:06:11,880 THESE ARE NOT INDIVIDUAL 1824 01:06:11,880 --> 01:06:12,800 MUTATIONS THESE ARE COLLECTIONS 1825 01:06:12,800 --> 01:06:14,800 OF MUTATIONS THAT SEEM TO 1826 01:06:14,800 --> 01:06:15,760 CO-OCCUR, AND I'LL BET YOU 1827 01:06:15,760 --> 01:06:18,960 DOLLARS TO DONUTS THEN THAT WHEN 1828 01:06:18,960 --> 01:06:20,440 WE DEFINE A GENETIC SUBTYPE, 1829 01:06:20,440 --> 01:06:23,600 BASICALLY WHAT WE'RE SEEING IN 1830 01:06:23,600 --> 01:06:25,040 AN INDIRECT WAY IS A DIFFERENT 1831 01:06:25,040 --> 01:06:28,960 CLASS OF MASTER REGULATORS THAT 1832 01:06:28,960 --> 01:06:30,520 GOES TO THAT SUBTYPE, BUT IN THE 1833 01:06:30,520 --> 01:06:31,680 PAPER WE JUST PUBLISHED IN 1834 01:06:31,680 --> 01:06:35,720 CANCER CELL, THESE THINGS HAVE A 1835 01:06:35,720 --> 01:06:39,400 100% PREDICTIVE POWER FOR 1836 01:06:39,400 --> 01:06:43,200 RESPONSE TO ABRUTE ANYBODY -- OR 1837 01:06:43,200 --> 01:06:47,960 BENEFIT FROM ABRUTANIB -- SO IT 1838 01:06:47,960 --> 01:06:49,800 MAY BE THAT WE HAVEN'T BEEN 1839 01:06:49,800 --> 01:06:50,680 SOPHISTICATED ENOUGH ABOUT 1840 01:06:50,680 --> 01:06:53,600 UNDERSTANDING HOW TO THINK ABOUT 1841 01:06:53,600 --> 01:06:56,440 THESE MUTATIONS AND MAYBE 1842 01:06:56,440 --> 01:06:58,240 THERE'S A NICE -- THERE'S A NICE 1843 01:06:58,240 --> 01:07:00,280 POINT IN THE MIDDLE THAT TAKES 1844 01:07:00,280 --> 01:07:04,320 INTO ACCOUNT BOTH OF THOSE BITS 1845 01:07:04,320 --> 01:07:04,680 OF INFORMATION. 1846 01:07:04,680 --> 01:07:06,120 >> I THINK THERE'S A RATIONALE 1847 01:07:06,120 --> 01:07:07,600 FOR THAT, LOU. 1848 01:07:07,600 --> 01:07:08,680 I THINK THERE'S TWO THINGS. 1849 01:07:08,680 --> 01:07:10,960 FIRST OF ALL, YOUR WORK WHICH IS 1850 01:07:10,960 --> 01:07:12,080 EXTREMELY SOPHISTICATED COULD 1851 01:07:12,080 --> 01:07:13,960 NOT HAVE HAPPENED WITHOUT VERY, 1852 01:07:13,960 --> 01:07:17,920 VERY PRECISE PATHWAYS, SIGNALING 1853 01:07:17,920 --> 01:07:20,440 PATHWAYS BUILT BY -- OVER -- 1854 01:07:20,440 --> 01:07:20,640 YEARS. 1855 01:07:20,640 --> 01:07:21,080 >> ABSOLUTELY. 1856 01:07:21,080 --> 01:07:22,960 >> SO YOU'RE ACTUALLY BENEFITING 1857 01:07:22,960 --> 01:07:24,000 FROM SOMETHING THAT ALMOST 1858 01:07:24,000 --> 01:07:25,200 DOESN'T EXIST IN OTHER TUMOR 1859 01:07:25,200 --> 01:07:25,400 TYPES. 1860 01:07:25,400 --> 01:07:26,000 >> TRUE. 1861 01:07:26,000 --> 01:07:27,240 >> SO BASICALLY YOU'RE DOING 1862 01:07:27,240 --> 01:07:28,560 WHAT WE'RE DOING BUT YOU'RE 1863 01:07:28,560 --> 01:07:29,800 USING THE LITERATURE TO -- 1864 01:07:29,800 --> 01:07:31,040 >> THE KNOWLEDGE BASE. 1865 01:07:31,040 --> 01:07:37,320 >> AND YOU -- BUT I FIND THIS 1866 01:07:37,320 --> 01:07:39,440 EXACTLY COALESCE INTO A PATHWAY, 1867 01:07:39,440 --> 01:07:40,640 ALL THE MUTATION IN THAT 1868 01:07:40,640 --> 01:07:42,000 PATHWAY, YOU CAN PREDICT. 1869 01:07:42,000 --> 01:07:46,680 THAT'S GREAT. 1870 01:07:46,680 --> 01:07:48,680 WHERE EVERY TUMOR HAS THE SAME 1871 01:07:48,680 --> 01:07:53,920 MAIN MUTATION IS -- AND THE 1872 01:07:53,920 --> 01:07:56,680 OTHER MUTATION ARE JUST 1873 01:07:56,680 --> 01:07:59,200 COMPLETELY COMPREHENSIBLE OR 1874 01:07:59,200 --> 01:07:59,720 MOSTLY COMPREHENSIBLE, YOU 1875 01:07:59,720 --> 01:08:01,600 REALLY DON'T KNOW WHAT TO DO, 1876 01:08:01,600 --> 01:08:02,840 YOU CANNOT USE THE APPROACH 1877 01:08:02,840 --> 01:08:04,000 YOU'VE DEVELOPED, LOU. 1878 01:08:04,000 --> 01:08:05,600 SO I THINK THIS IS WHERE THE 1879 01:08:05,600 --> 01:08:06,640 COMMON GROUND EXISTS BETWEEN 1880 01:08:06,640 --> 01:08:10,040 WHAT WE DO AND WHAT YOU HAVE 1881 01:08:10,040 --> 01:08:11,160 DONE. 1882 01:08:11,160 --> 01:08:12,640 TO SOME EXTENT, WE'VE TAKEN A 1883 01:08:12,640 --> 01:08:14,000 PAGE FROM YOUR BOOK AND THE WAY 1884 01:08:14,000 --> 01:08:15,280 WE HAVE STARTED TO DEVELOP THESE 1885 01:08:15,280 --> 01:08:16,160 THINGS PRECISELY -- 1886 01:08:16,160 --> 01:08:18,400 >> BUT JUST FOR THE AUDIENCE, 1887 01:08:18,400 --> 01:08:20,040 AND WE'LL STOP IT THERE, WHEN 1888 01:08:20,040 --> 01:08:21,760 I'M SAYING A GENETIC SUBTYPE, I 1889 01:08:21,760 --> 01:08:23,400 THINK YOU SHOULD SORT OF THINK 1890 01:08:23,400 --> 01:08:26,800 OF, YOU KNOW, LUMINAL BREAST 1891 01:08:26,800 --> 01:08:29,600 CANCER VERSUS BASAL, THAT THAT'S 1892 01:08:29,600 --> 01:08:32,960 THE KIND OF BIG DISTINCTION, AND 1893 01:08:32,960 --> 01:08:34,600 THAT DOES TRACK WITH A BUNCH OF 1894 01:08:34,600 --> 01:08:35,800 DIFFERENT MASTER REGULATOR 1895 01:08:35,800 --> 01:08:37,240 DIFFERENCES BETWEEN THOSE TWO. 1896 01:08:37,240 --> 01:08:39,120 SO IT'S NOT JUST ONE PATHWAY, 1897 01:08:39,120 --> 01:08:44,160 IT'S A WHOLE CONSTELLATION OF -- 1898 01:08:44,160 --> 01:08:49,040 IT'S SORT OF LIKE A SELIB ORIGIN 1899 01:08:49,040 --> 01:08:50,560 BASED TYPE OF CANCER THAT SEEMED 1900 01:08:50,560 --> 01:08:52,440 TO TRACK WITH THERAPEUTIC 1901 01:08:52,440 --> 01:08:53,440 RESPONSE, AND WHAT I'D LOVE TO 1902 01:08:53,440 --> 01:08:54,640 DO WITH YOU COLLABORATIVELY IS 1903 01:08:54,640 --> 01:08:56,800 TO SEE IF IT TRACKS WITH MASTER 1904 01:08:56,800 --> 01:08:57,280 REGULATORS. 1905 01:08:57,280 --> 01:08:58,520 SO WE'LL LEAVE IT AT THAT. 1906 01:08:58,520 --> 01:09:02,800 I HOPE THIS WAS -- I HOPE THE 1907 01:09:02,800 --> 01:09:07,280 LAST QUESTION AND ANSWER WAS 1908 01:09:07,280 --> 01:09:07,840 ENTERTAINING FOR EVERYBODY. 1909 01:09:07,840 --> 01:09:10,200 BUT THANK YOU, ANDREA. 1910 01:09:10,200 --> 01:09:11,480 I ALWAYS LEARN A LOT, AND THIS 1911 01:09:11,480 --> 01:09:12,600 WAS ANOTHER GREAT LECTURE. 1912 01:09:12,600 --> 01:09:13,760 THANK YOU SO MUCH. 1913 01:09:13,760 --> 00:00:00,000 >> THANK YOU, LOU.