1 00:00:05,280 --> 00:00:09,600 >>IT'S VERY NICE TO SEE SOME 2 00:00:09,600 --> 00:00:10,680 LEVEL OF IN PERSON CROWD. 3 00:00:10,680 --> 00:00:17,600 I THINK I GAVE A TALK HERE A 4 00:00:17,600 --> 00:00:19,280 COUPLE MONTHS AGO AND THIS IS 5 00:00:19,280 --> 00:00:20,600 MUCH MORE PEOPLE HERE SO THANKS 6 00:00:20,600 --> 00:00:22,880 FOR COMING. 7 00:00:22,880 --> 00:00:25,240 IT GIVES ME GREAT PLEASURE TO 8 00:00:25,240 --> 00:00:27,640 INTRODUCE LAZZERINI DENCHI AS 9 00:00:27,640 --> 00:00:29,240 OUR CCR GRAND ROUND SPEAKER. 10 00:00:29,240 --> 00:00:34,000 HE JOINED CCR IN 2018 AS A 11 00:00:34,000 --> 00:00:35,320 STADTMAN INVESTIGATOR AND BECAME 12 00:00:35,320 --> 00:00:37,520 SENIOR INVESTIGATOR IN 2021. 13 00:00:37,520 --> 00:00:39,960 HE GOT TO A LIGHTNING START AND 14 00:00:39,960 --> 00:00:42,720 HAS BEEN AWARDED THE NCI 15 00:00:42,720 --> 00:00:43,320 RESEARCH HIGHLIGHT AWARD IN 16 00:00:43,320 --> 00:00:45,160 2022. 17 00:00:45,160 --> 00:00:49,640 AND THE NCI DIRECTOR'S AWARD FOR 18 00:00:49,640 --> 00:00:52,480 BASIC BIOLOGICAL SCIENCES IN 19 00:00:52,480 --> 00:00:53,840 2023. 20 00:00:53,840 --> 00:00:57,360 CONGRATULATIONS TO EROS. 21 00:00:57,360 --> 00:00:59,480 BEFORE COMING TO BETHESDA HE WAS 22 00:00:59,480 --> 00:01:02,240 AN INVESTIGATOR AT THE SCRIPPS 23 00:01:02,240 --> 00:01:11,000 INSTITUTE IN LA JOLLA AFTER IT 24 00:01:11,000 --> 00:01:11,240 YEARS. 25 00:01:11,240 --> 00:01:13,520 THE WORKED IN LAB SPIKED HIS 26 00:01:13,520 --> 00:01:14,960 INTEREST IN TELOMERE BIOLOGY AND 27 00:01:14,960 --> 00:01:17,800 CONTINUES IN THIS AREA TODAY. 28 00:01:17,800 --> 00:01:20,000 THE STUDY OF TELOMERE REGULATION 29 00:01:20,000 --> 00:01:22,040 SAY PRETTY CROWDED FIELD BECAUSE 30 00:01:22,040 --> 00:01:25,040 IT HAS BROAD IMPLICATIONS FOR 31 00:01:25,040 --> 00:01:26,960 AGING, SENESCENCE, CANCER AND 32 00:01:26,960 --> 00:01:37,480 CLINICAL IMPLICATIONS BUT EROS 33 00:01:40,520 --> 00:01:41,600 HAS DISCOVERED REPRESSED 34 00:01:41,600 --> 00:01:44,520 ACTIVATION AT THE NATIONAL 35 00:01:44,520 --> 00:01:48,000 CHROMOSOME ENDS, DISCOVERY OF 36 00:01:48,000 --> 00:01:52,040 TELOMERE PROTEINS AND LENGTH AND 37 00:01:52,040 --> 00:01:55,280 THE REQUIREMENT FOR TELOMERE 38 00:01:55,280 --> 00:01:57,800 PROTEINS IN STEM CELLS VERSUS 39 00:01:57,800 --> 00:01:58,720 DIFFERENTIATED CELLS. 40 00:01:58,720 --> 00:02:02,760 HE WAS DOING THIS FANTASTIC WORK 41 00:02:02,760 --> 00:02:04,840 AT SCRIPPS I FOLLOWED CLOSELY 42 00:02:04,840 --> 00:02:06,320 AND AS SOON AS WE CONVINCED HIM 43 00:02:06,320 --> 00:02:08,680 TO COME TO NIH AND WE WERE HAPPY 44 00:02:08,680 --> 00:02:10,400 IT MAKE HIM AN OFFER AND HAPPY 45 00:02:10,400 --> 00:02:12,160 HE JOINED. 46 00:02:12,160 --> 00:02:13,400 AT NIH I'VE HAD OPPORTUNITY TO 47 00:02:13,400 --> 00:02:17,040 SEE FIRST-HAND HOW HE APPROACHES 48 00:02:17,040 --> 00:02:18,760 HIS -- HOW HE RUNS HIS LAB AND 49 00:02:18,760 --> 00:02:20,880 APPROACHES HIS WORK. 50 00:02:20,880 --> 00:02:23,360 TO GIVE AN EXAMPLE, HE RECENTLY 51 00:02:23,360 --> 00:02:25,880 MADE AN UNEXPECTED FINDING ABOUT 52 00:02:25,880 --> 00:02:28,320 TELOMERE END PROTECTION WHICH 53 00:02:28,320 --> 00:02:30,440 WENT I THINK AGAINST THE DOGMA 54 00:02:30,440 --> 00:02:33,640 THAT'S BEEN AROUND MORE THAN 25 55 00:02:33,640 --> 00:02:35,040 YEARS WHICH I'M SURE HE'LL 56 00:02:35,040 --> 00:02:35,280 MENTION. 57 00:02:35,280 --> 00:02:37,280 AT THE SAME TIME HE FOUND OUT 58 00:02:37,280 --> 00:02:38,840 ANOTHER COLLEAGUE OF HIS HAD 59 00:02:38,840 --> 00:02:40,680 MADE A SIMILAR FINDING. 60 00:02:40,680 --> 00:02:42,640 SO THEY WERE CONFIDENT THE 61 00:02:42,640 --> 00:02:44,280 FINDINGS WERE TRUE. 62 00:02:44,280 --> 00:02:46,280 THE ONLY PROBLEM WAS FIGURING 63 00:02:46,280 --> 00:02:47,440 OUT THE MECHANISM. 64 00:02:47,440 --> 00:02:48,240 THAT'S THE HARD PART. 65 00:02:48,240 --> 00:02:57,280 SO EROS DECIDING HE PREFERRED TO 66 00:02:57,280 --> 00:02:59,720 CRACK THE PROBLEM HIMSELF AND 67 00:02:59,720 --> 00:03:00,680 THAT'S WHERE THE FUN WAS WHERE 68 00:03:00,680 --> 00:03:02,560 HE DID ELEGANTLY AND I THINK 69 00:03:02,560 --> 00:03:03,400 THEY CAME UP WITH SLIGHTLY 70 00:03:03,400 --> 00:03:06,000 DIFFERENT ANSWERS AS WELL. 71 00:03:06,000 --> 00:03:09,840 AND THIS IS PRECISELY WHAT I 72 00:03:09,840 --> 00:03:15,920 FOUND TO BE HIS MODUS OPERANDI 73 00:03:15,920 --> 00:03:20,120 AND STICKS TO HIS GUN AND ENJOYS 74 00:03:20,120 --> 00:03:21,600 SOLVING SCIENTIFIC PROBLEMS 75 00:03:21,600 --> 00:03:22,760 LEADING TO ELEGANT SOLUTIONS. 76 00:03:22,760 --> 00:03:24,240 AT THE SAME TIME HE'S BEEN A 77 00:03:24,240 --> 00:03:34,680 PLEASURE TO WORK WITH, A 78 00:03:40,560 --> 00:03:41,640 TERRIFIC COLLABORATOR AND 79 00:03:41,640 --> 00:03:43,800 ENCOURAGE YOU TO TALK TO HIM 80 00:03:43,800 --> 00:03:44,640 AFTER HIS GRAND ROUNDS SO TALK 81 00:03:44,640 --> 00:03:44,840 TO HIM. 82 00:03:44,840 --> 00:03:50,680 THANKS. 83 00:03:50,680 --> 00:03:53,280 >>THANK YOU. 84 00:03:53,280 --> 00:03:53,840 THAT WAS NICE. 85 00:03:53,840 --> 00:03:56,520 IT WAS A KIND INTRODUCTION. 86 00:03:56,520 --> 00:03:59,280 IT'S BEEN A PLEASURE TO JOIN 87 00:03:59,280 --> 00:03:59,520 NCI. 88 00:03:59,520 --> 00:04:00,480 IT'S BEEN A PHENOMENAL TIME TO 89 00:04:00,480 --> 00:04:02,400 BE HERE. 90 00:04:02,400 --> 00:04:11,080 IT'S BEEN NICE TO WORK WITH 91 00:04:11,080 --> 00:04:18,320 AND-DRE AND EVERYONE IN THE 92 00:04:18,320 --> 00:04:21,000 BRANCH AND I'LL TALK ABOUT 93 00:04:21,000 --> 00:04:22,680 TELOMERES AND TELL STORIES IN 94 00:04:22,680 --> 00:04:23,760 WHAT WE'RE TRYING TO UNDERSTAND 95 00:04:23,760 --> 00:04:25,800 WHEN TELOMERES AND CANCER. 96 00:04:25,800 --> 00:04:27,720 MAYBE I WAS AMBITIOUS WITH THE 97 00:04:27,720 --> 00:04:28,280 DEVELOPMENT PART. 98 00:04:28,280 --> 00:04:30,000 YOU'LL TELL ME ABOUT IT. 99 00:04:30,000 --> 00:04:32,320 FIRST OF ALL, WHAT ARE 100 00:04:32,320 --> 00:04:32,720 TELOMERES? 101 00:04:32,720 --> 00:04:35,000 THIS IS THE WAY EVERY MAMMAL 102 00:04:35,000 --> 00:04:37,560 STUDIED SO FAR LOOKS LIKE AT THE 103 00:04:37,560 --> 00:04:38,560 CHROMOSOMAL END. 104 00:04:38,560 --> 00:04:44,000 EACH CHROMOSOME END AND I'LL TRY 105 00:04:44,000 --> 00:04:50,200 TO DO THIS -- SO EVERY SINGLE 106 00:04:50,200 --> 00:04:54,400 MAMMAL HAS THE SAME REPETITIVE 107 00:04:54,400 --> 00:04:57,440 SEQUENCE AT EACH CHROMOSOME END 108 00:04:57,440 --> 00:04:58,360 AND AT THE END THERE'S A STRETCH 109 00:04:58,360 --> 00:05:03,080 OF SINGLE-STRANDED DNA. 110 00:05:03,080 --> 00:05:05,040 AND THE ONLY DIFFERENCE BETWEEN 111 00:05:05,040 --> 00:05:06,000 THE DIFFERENT SPECIES IN MAMMALS 112 00:05:06,000 --> 00:05:07,640 IS THE LENGTHS. 113 00:05:07,640 --> 00:05:14,560 IN HUMANS WE HAVE ROUGHLY AATC 114 00:05:14,560 --> 00:05:16,520 AND MICE HAVE DIFFERENT 115 00:05:16,520 --> 00:05:21,040 TELOMERES BUT THE OVER ALL 116 00:05:21,040 --> 00:05:23,800 STRUCTURE IS THE SAME AND FOR 117 00:05:23,800 --> 00:05:25,040 MAMMALS WITH A SLIGHTLY 118 00:05:25,040 --> 00:05:27,080 DIFFERENT SEQUENCE FOLLOW THE 119 00:05:27,080 --> 00:05:28,240 SAME T. 120 00:05:28,240 --> 00:05:29,720 THE DIFFERENCE IS RELATED TO 121 00:05:29,720 --> 00:05:31,880 WHAT TELOMERES DO. 122 00:05:31,880 --> 00:05:33,480 THEY SOLVE TWO PROBLEMS 123 00:05:33,480 --> 00:05:34,720 ASSOCIATED WITH LINEAR 124 00:05:34,720 --> 00:05:35,080 CHROMOSOMES. 125 00:05:35,080 --> 00:05:38,200 WHAT I FIND REMARKABLE IS THE 126 00:05:38,200 --> 00:05:40,360 FACT THAT THESE PROBLEMS WERE 127 00:05:40,360 --> 00:05:41,840 DISCOVERED BEFORE WE KNEW 128 00:05:41,840 --> 00:05:42,960 ANYTHING ABOUT TELOMERES. 129 00:05:42,960 --> 00:05:44,920 THE FIRST PROBLEM HAS TO DO WITH 130 00:05:44,920 --> 00:05:48,000 THE SO-CALLED END REPLICATION 131 00:05:48,000 --> 00:05:48,200 PROBLEM. 132 00:05:48,200 --> 00:05:50,200 SO WHAT IS THAT? 133 00:05:50,200 --> 00:05:52,960 THE END REPLICATION PROBLEM IS 134 00:05:52,960 --> 00:05:55,320 THE FACT OUR DNA REPLICATION 135 00:05:55,320 --> 00:05:57,160 WORKS. 136 00:05:57,160 --> 00:05:59,560 WHEN YOU TRY TO DNA MACHINERY 137 00:05:59,560 --> 00:06:02,480 TRIES TO COPY A LINEAR TEMPLATE, 138 00:06:02,480 --> 00:06:04,280 THERE'S ALWAYS GOING TO BE A GAP 139 00:06:04,280 --> 00:06:05,760 OF INFORMATION THAT IS LOST. 140 00:06:05,760 --> 00:06:08,320 AND IT'S DUE TO THE FACT THAT IN 141 00:06:08,320 --> 00:06:13,320 THE LIGAND STRENGTH NOW MATTER 142 00:06:13,320 --> 00:06:15,360 HOW YOU PUT IT TO THE TIP OF THE 143 00:06:15,360 --> 00:06:17,520 DNA THAT PORTION OF INFORMATION 144 00:06:17,520 --> 00:06:20,280 WILL BE LOST. 145 00:06:20,280 --> 00:06:26,840 SO WHEN A CELL DIVIDES A PORTION 146 00:06:26,840 --> 00:06:28,840 WILL LOST AND IT WILL LEAD TO 147 00:06:28,840 --> 00:06:29,400 CELL DEATH. 148 00:06:29,400 --> 00:06:31,520 OF COURSE, WE DON'T DIE, WE'RE 149 00:06:31,520 --> 00:06:34,600 STICKING AROUND AND THE REASON 150 00:06:34,600 --> 00:06:37,240 IS WE HAVE WAYS TO DEAL WITH THE 151 00:06:37,240 --> 00:06:37,480 PROBLEM. 152 00:06:37,480 --> 00:06:40,720 FOR THE MOST PART THE MOST 153 00:06:40,720 --> 00:06:43,160 FREQUENT WAY WE DEAL WITH IT IS 154 00:06:43,160 --> 00:06:50,600 BY AN ENZYME CALLED TELEMERASE 155 00:06:50,600 --> 00:06:53,080 WHICH ADDS NEW REPEAT USING ITS 156 00:06:53,080 --> 00:06:56,200 OWN RNA AS A TEMPLATE AND WILL 157 00:06:56,200 --> 00:07:01,240 ADD NEW SEQUENCES THEREFORE 158 00:07:01,240 --> 00:07:06,360 COUNTER ACTING THE LOST 159 00:07:06,360 --> 00:07:06,720 INFORMATION. 160 00:07:06,720 --> 00:07:08,320 THIS IS THE HOW OUR CELLS DEAL 161 00:07:08,320 --> 00:07:09,200 WITH THE PROBLEM. 162 00:07:09,200 --> 00:07:13,680 AND THE SECOND PROBLEM IS MORE 163 00:07:13,680 --> 00:07:15,280 PRESSING WHILE THE REPLICATION 164 00:07:15,280 --> 00:07:18,800 PROBLEM IS ONLY A PROBLEM WHEN 165 00:07:18,800 --> 00:07:22,560 CELLS DIVIDE THE SECOND PROBLEM 166 00:07:22,560 --> 00:07:33,080 WITH TELOMERES WITH CELLS NOT 167 00:07:33,400 --> 00:07:33,880 DIVID 168 00:07:33,880 --> 00:07:34,160 DIVIDING. 169 00:07:34,160 --> 00:07:35,760 OUR CELLS DE GAP IN THE DAMAGE 170 00:07:35,760 --> 00:07:37,920 AND DAMAGE AND THIS WILL TRIGGER 171 00:07:37,920 --> 00:07:39,040 DNA DAMAGE RESPONSE. 172 00:07:39,040 --> 00:07:41,320 THE CELLS FIX IT AND IF THEY 173 00:07:41,320 --> 00:07:42,880 CANNOT THEY WILL KILL THE CELL. 174 00:07:42,880 --> 00:07:47,040 THIS IS GREAT FOR THE GENOME BUT 175 00:07:47,040 --> 00:07:48,960 BECOMES A PROBLEM WHEN YOU DEAL 176 00:07:48,960 --> 00:07:51,200 WITH CHROMOSOMAL ENDS IT'S A 177 00:07:51,200 --> 00:07:52,280 BROKEN PIECE OF DNA IN THE 178 00:07:52,280 --> 00:07:55,040 CHROMOSOME SO WE SHOULD BE 179 00:07:55,040 --> 00:07:57,960 INDUCING DNA DAMAGE ALL THE TIME 180 00:07:57,960 --> 00:08:00,280 IN EVERY CELL OF OUR BODY ALL 181 00:08:00,280 --> 00:08:01,880 THE TIME. 182 00:08:01,880 --> 00:08:03,960 WE DON'T DO IT AND IT'S BECAUSE 183 00:08:03,960 --> 00:08:05,880 THERE'S A SET OF PROTEINS THAT 184 00:08:05,880 --> 00:08:08,560 BIND TO CHROMOSOME ENDS AND THEY 185 00:08:08,560 --> 00:08:12,280 SORT OF SHIELD OR HIDE THE 186 00:08:12,280 --> 00:08:14,080 CHROMOSOME FROM THE DNA DAMAGE 187 00:08:14,080 --> 00:08:15,600 ACTIVITY PREVENTING ACTIVATION. 188 00:08:15,600 --> 00:08:17,880 THIS IS HOW YOU SOLVE THE END 189 00:08:17,880 --> 00:08:18,600 PROTECTION PROBLEM. 190 00:08:18,600 --> 00:08:20,800 I'LL SHOW AN EXAMPLE AND THIS IS 191 00:08:20,800 --> 00:08:22,760 MY FAVORITE EXAMPLE BECAUSE IT 192 00:08:22,760 --> 00:08:25,120 DEALS WITH MY FAVORITE PROTEIN 193 00:08:25,120 --> 00:08:27,640 IN TELOMERES THIS PROTEIN CALLED 194 00:08:27,640 --> 00:08:28,160 TR2. 195 00:08:28,160 --> 00:08:30,360 IN THIS CASE WHAT I'M SHOWING IS 196 00:08:30,360 --> 00:08:32,160 IN RED YOU HAVE CHROMOSOMES IN 197 00:08:32,160 --> 00:08:35,280 GREEN YOU CAN HAVE THE TTAG 198 00:08:35,280 --> 00:08:40,360 REPEAT AND TELOMERES AND THIS 199 00:08:40,360 --> 00:08:42,160 DERIVES FROM A MOUSE MODEL WE 200 00:08:42,160 --> 00:08:43,320 GENERATED THAT WAS GENERATED IN 201 00:08:43,320 --> 00:08:46,040 THE LAB WHEN I WAS DOING MY 202 00:08:46,040 --> 00:08:49,080 POSTDOC AND IN WHICH WE CAN 203 00:08:49,080 --> 00:08:51,120 DELETE CONTINUALLY TR2. 204 00:08:51,120 --> 00:08:54,960 IN THIS SITUATION TR2 IS 205 00:08:54,960 --> 00:09:01,120 EXPRESSED, IT'S COMPLETELY FINE 206 00:09:01,120 --> 00:09:09,400 BUT WE CAN DELETE TR2 AND I FELL 207 00:09:09,400 --> 00:09:11,320 IN LOVE WITH THE PROTEIN AND 208 00:09:11,320 --> 00:09:13,440 ASKED MYSELF HOW IT WORKS. 209 00:09:13,440 --> 00:09:15,760 BY REMOVING THE PROTEIN AND 210 00:09:15,760 --> 00:09:19,320 GIVING IT ONE SINGLE CELL CYCLE 211 00:09:19,320 --> 00:09:21,280 EACH CHROMOSOME IS RECOGNIZED AS 212 00:09:21,280 --> 00:09:25,720 A SITE OF DNA DAMAGE AND FUSED 213 00:09:25,720 --> 00:09:27,120 TOGETHER TO REPAIR. 214 00:09:27,120 --> 00:09:30,720 THIS IS THE BEST EXPLANATION AND 215 00:09:30,720 --> 00:09:33,760 BEST EXAMPLE OF END PROTECTION. 216 00:09:33,760 --> 00:09:38,760 THE FUNCTION THIS PROTEINS DO AT 217 00:09:38,760 --> 00:09:39,400 TELOMERES. 218 00:09:39,400 --> 00:09:41,560 ZOOMING OUT AND I WON'T GO INTO 219 00:09:41,560 --> 00:09:43,200 ALL DETAILS AND THE WAY I SEE 220 00:09:43,200 --> 00:09:45,000 ALL PROTEINS THAT BIND TO 221 00:09:45,000 --> 00:09:47,120 CHROMOSOME ENDS THEY DEAL WITH 222 00:09:47,120 --> 00:09:48,480 ALL THE DIFFERENT TYPES OF DNA 223 00:09:48,480 --> 00:09:50,280 DAMAGE PRESENT IN A CELL. 224 00:09:50,280 --> 00:09:51,560 YOU HAVE SPECIALIZED PROTEINS 225 00:09:51,560 --> 00:09:52,640 THAT CAN SUPPRESS DIFFERENT 226 00:09:52,640 --> 00:09:53,880 TYPES OF DNA DAMAGE. 227 00:09:53,880 --> 00:09:56,520 THE RESULT IS OUR CHROMOSOME 228 00:09:56,520 --> 00:10:00,720 ENDS ARE NOT STICKING. 229 00:10:00,720 --> 00:10:06,360 THEY DON'T BECOME ENGAGED AB AND 230 00:10:06,360 --> 00:10:08,480 THIS IS A FASCINATING PROBLEM 231 00:10:08,480 --> 00:10:11,200 BUT THERE'S CLEAR IMPLICATION 232 00:10:11,200 --> 00:10:12,560 FOR HUMAN HEALTH AND ESPECIALLY 233 00:10:12,560 --> 00:10:13,320 FOR TUMORS. 234 00:10:13,320 --> 00:10:16,200 THAT'S WHAT I'M GOING TO FOCUS 235 00:10:16,200 --> 00:10:16,440 TODAY. 236 00:10:16,440 --> 00:10:19,240 SO THE VERY FIRST PROBLEM I 237 00:10:19,240 --> 00:10:21,760 INTRODUCED THE END REPLICATION 238 00:10:21,760 --> 00:10:24,480 PROBLEM IS A STRONG IMPLICATION 239 00:10:24,480 --> 00:10:27,600 FOR TUMOR DEVELOPMENT BECAUSE IT 240 00:10:27,600 --> 00:10:29,520 REPRESENTS A MAJOR TUMOR 241 00:10:29,520 --> 00:10:31,560 SUPPRESSIVE PATHWAY. 242 00:10:31,560 --> 00:10:33,880 EACH CELL -- EACH CANCER IN 243 00:10:33,880 --> 00:10:36,120 HUMANS NEEDS TO OVERCOME THE END 244 00:10:36,120 --> 00:10:37,680 REPLICATION PROBLEM. 245 00:10:37,680 --> 00:10:43,160 THE REASON IS HUMANS DON'T 246 00:10:43,160 --> 00:10:46,840 EXPRESS THE TELOMERASE SO FOR A 247 00:10:46,840 --> 00:10:50,200 TUMOR TO BE SUCCESSFUL AND GROW 248 00:10:50,200 --> 00:10:54,680 NEEDS TO OVERCOME THE IMPULSE BY 249 00:10:54,680 --> 00:10:55,920 TEL MERE SHORTENING. 250 00:10:55,920 --> 00:10:58,440 THIS IS OUR OWN MARKER OF TUMOR 251 00:10:58,440 --> 00:10:58,720 BIOLOGY. 252 00:10:58,720 --> 00:11:01,360 THEN WAY THEY DO IS IT BY THE 253 00:11:01,360 --> 00:11:07,280 MOST PART BY REACTIVATING 254 00:11:07,280 --> 00:11:08,720 TELOMERASE AND THERE'S A 255 00:11:08,720 --> 00:11:11,200 PROMOTION THAT ALLOWS FOR 256 00:11:11,200 --> 00:11:12,840 REACTIVATION AND IT OCCURS IN 257 00:11:12,840 --> 00:11:15,920 ROUGHLY 80% TO 90% OF HUMAN 258 00:11:15,920 --> 00:11:16,160 TUMORS. 259 00:11:16,160 --> 00:11:18,400 HOWEVER, THERE'S ANOTHER SEFT 260 00:11:18,400 --> 00:11:20,200 TUMORS THAT DON'T EXPRESS 261 00:11:20,200 --> 00:11:24,520 TELOMERES AND THESE INSTEAD USE 262 00:11:24,520 --> 00:11:27,720 THE L PATHWAY WITH A TELOMERE 263 00:11:27,720 --> 00:11:29,080 ELONGATION WE'LL GO IN THE 264 00:11:29,080 --> 00:11:30,920 DETAILS TO AFTERWARDS AND 265 00:11:30,920 --> 00:11:32,920 REPRESENT ROUGHLY 50% OF TUMORS. 266 00:11:32,920 --> 00:11:34,400 WHAT IS INTERESTING IS NOT ALL 267 00:11:34,400 --> 00:11:37,680 TUMORS ARE THE SAME WHEN IT 268 00:11:37,680 --> 00:11:40,920 COMES TO TELOMERE MAINTENANCE 269 00:11:40,920 --> 00:11:43,320 AND THE TUMORS OF THESE ORIGINS 270 00:11:43,320 --> 00:11:45,880 INCLUDING SARCOMAS SEND TO HAVE 271 00:11:45,880 --> 00:11:47,240 HIGH FREQUENCY OF RELIANCE ON 272 00:11:47,240 --> 00:11:50,000 THIS PATHWAY WHICH IS A PATHWAY 273 00:11:50,000 --> 00:11:53,760 WE DON'T UNDERSTAND VERY WELL. 274 00:11:53,760 --> 00:11:53,960 OKAY. 275 00:11:53,960 --> 00:11:57,160 SO THE SECOND PROBLEM THAT I 276 00:11:57,160 --> 00:11:59,720 INTRODUCED THE END PROTECTION 277 00:11:59,720 --> 00:12:01,840 ALSO AS IMPORTANCE DURING TUMOR 278 00:12:01,840 --> 00:12:03,840 DEVELOPMENT AND THIS HAS TO DO 279 00:12:03,840 --> 00:12:05,680 WITH GENOME INSTABILITY. 280 00:12:05,680 --> 00:12:07,360 IT'S BEEN SHOWN IN MANY SYSTEMS 281 00:12:07,360 --> 00:12:09,280 INCLUDING MOUSE MODELS AS WELL 282 00:12:09,280 --> 00:12:12,640 AS IN CLINICAL STUDIES THAT WHEN 283 00:12:12,640 --> 00:12:14,360 THERE IS -- WHEN TELOMERES 284 00:12:14,360 --> 00:12:15,080 BECOME VERY SHORT THERE'S A 285 00:12:15,080 --> 00:12:18,320 PHASE IN WHICH THEY CAN GROW 286 00:12:18,320 --> 00:12:20,520 WITHOUT ANY INDUCED TELOMERE 287 00:12:20,520 --> 00:12:20,880 ELONGATION. 288 00:12:20,880 --> 00:12:25,160 IF THAT HAPPENS, TELOMERES ARE A 289 00:12:25,160 --> 00:12:28,280 DRIVER FOR INSTABILITY AND ONE 290 00:12:28,280 --> 00:12:32,000 NICE EXAMPLE IS PROBABLY ONE OF 291 00:12:32,000 --> 00:12:34,760 THE MOST CATASTROPHIC GENE 292 00:12:34,760 --> 00:12:39,120 REARRANGEMENT SEEN IN TUMORS AND 293 00:12:39,120 --> 00:12:40,920 WHERE YOU SEE CANCER CELLS 294 00:12:40,920 --> 00:12:46,640 TRIGGER IS BY END TO END 295 00:12:46,640 --> 00:12:48,320 CHROMOSOMAL FUSIONS. 296 00:12:48,320 --> 00:12:52,720 SO HOPEFULLY I CONVINCED WE ARE 297 00:12:52,720 --> 00:12:57,760 STUDYING THIS HAS I MPLICATIONS 298 00:12:57,760 --> 00:12:58,840 FOR CANCER BIOLOGY. 299 00:12:58,840 --> 00:13:02,600 I'LL TELL YOU TWO STORIES OR TWO 300 00:13:02,600 --> 00:13:03,400 ASPECTS OF OUR RESEARCH. 301 00:13:03,400 --> 00:13:04,680 BY DOING SO HOPEFULLY I CAN SHOW 302 00:13:04,680 --> 00:13:06,400 YOU WHAT WE DO AND HOPEFULLY 303 00:13:06,400 --> 00:13:12,520 ENGAGE POTENTIAL COLLABORATION. 304 00:13:12,520 --> 00:13:15,480 THE FIRST STORY HAS TO DO WITH 305 00:13:15,480 --> 00:13:16,720 HOW IT BYPASSES TUMOR 306 00:13:16,720 --> 00:13:18,160 SUPPRESSION. 307 00:13:18,160 --> 00:13:21,120 WHAT I MEAN IS SO TELOMERE 308 00:13:21,120 --> 00:13:22,600 SHORTEN AS A TOLD YOU AND 309 00:13:22,600 --> 00:13:25,280 EVENTUALLY THEY BECOME SO SHORT 310 00:13:25,280 --> 00:13:26,480 IT CAN INDUCE DNA DAMAGE 311 00:13:26,480 --> 00:13:26,760 RESPONSE. 312 00:13:26,760 --> 00:13:28,280 IF IT WAS PERFECT THERE WOULD BE 313 00:13:28,280 --> 00:13:29,760 NO TUMORS. 314 00:13:29,760 --> 00:13:30,440 UNFORTUNATELY WE KNOW THAT'S NOT 315 00:13:30,440 --> 00:13:32,360 THE CASE. 316 00:13:32,360 --> 00:13:35,320 THERE IS A QUESTION HERE, WHY 317 00:13:35,320 --> 00:13:36,240 SOME CELLS CAN SURVIVE. 318 00:13:36,240 --> 00:13:40,520 WHAT ARE MECHANISMS THAT ALLOW 319 00:13:40,520 --> 00:13:43,680 CELLS TO SURVIVE AND ENGAGING 320 00:13:43,680 --> 00:13:45,240 THE DIFFERENT PATHWAY. 321 00:13:45,240 --> 00:13:46,440 THERE'S CLEARLY SOMETHING HERE 322 00:13:46,440 --> 00:13:47,520 WE DON'T UNDERSTAND. 323 00:13:47,520 --> 00:13:49,720 IT'S QUITE DIFFICULT TO STUDY IN 324 00:13:49,720 --> 00:13:52,480 VIVO BECAUSE CLEARLY MOST OF THE 325 00:13:52,480 --> 00:13:55,840 CELLS THE TUMOR CELLS THE 326 00:13:55,840 --> 00:13:57,240 PRECANCEROUS CELLS DIE OFF SO WE 327 00:13:57,240 --> 00:14:00,320 DON'T SEE THE RESULT OF THOSE 328 00:14:00,320 --> 00:14:05,520 AND THE ONE THAT CANNOT HAS 329 00:14:05,520 --> 00:14:07,360 TELOMERASE REACTIVATION AND WE 330 00:14:07,360 --> 00:14:08,800 LOOK AT WHAT WE CAN RESPONSE THE 331 00:14:08,800 --> 00:14:09,240 FUNCTION. 332 00:14:09,240 --> 00:14:14,880 THE WAY WE DO IT INSTEAD OF 333 00:14:14,880 --> 00:14:23,720 DRIVING THE SORTENING IS MESSING 334 00:14:23,720 --> 00:14:31,640 WITH THE TELOMERES THAT BIND AND 335 00:14:31,640 --> 00:14:33,680 MAYBE DIFFERENTIATION AND MAYBE 336 00:14:33,680 --> 00:14:36,720 WE CAN LOOK AT THE WAY THE CELLS 337 00:14:36,720 --> 00:14:38,640 RESPOND AND IT WAS MORE OF AN 338 00:14:38,640 --> 00:14:43,560 IDEA AND WHEN I JOINED NCI I WAS 339 00:14:43,560 --> 00:14:47,200 LUCKY TO RECRUIT MY FIRST 340 00:14:47,200 --> 00:14:49,360 POSTDOC THAT TURNED OUT TO BE AN 341 00:14:49,360 --> 00:14:54,280 AMAZING POSTDOC AND MARTA 342 00:14:54,280 --> 00:14:57,600 DECIDED TO TAKE ON THE PROJECT 343 00:14:57,600 --> 00:15:00,480 AND SOUGHT COLLABORATION WITH A 344 00:15:00,480 --> 00:15:05,640 LAB THAT STARTED ROUGHLY THE 345 00:15:05,640 --> 00:15:08,160 SAME TIME AND LOOKING AT THE 346 00:15:08,160 --> 00:15:11,880 CELLS AND DIFFERENTIATION AND 347 00:15:11,880 --> 00:15:13,360 LOOKED AT MICE CELLS SIMILAR TO 348 00:15:13,360 --> 00:15:15,040 THE MODEL I SHOWED YOU BEFORE 349 00:15:15,040 --> 00:15:20,720 WHEN YOU SAW THE CHROMOSOME 350 00:15:20,720 --> 00:15:21,120 FUSIONS. 351 00:15:21,120 --> 00:15:31,560 SO THE CELLS ARE PLURIPOTENT 352 00:15:31,560 --> 00:15:34,320 STEM CELLS AND SO MARTA DERIVED 353 00:15:34,320 --> 00:15:36,520 SOME DIFFERENTIATION FOR THE 354 00:15:36,520 --> 00:15:37,920 SOMATIC CELLS WE CALLED 355 00:15:37,920 --> 00:15:38,800 FIBROBLAST LIKE CELLS BECAUSE WE 356 00:15:38,800 --> 00:15:41,160 DON'T KNOW WHAT THEY ARE BUT 357 00:15:41,160 --> 00:15:43,080 THEY LOOK LIKE FIBROBLASTS AND 358 00:15:43,080 --> 00:15:45,240 EXPRESS GENES EXPRESSED IN 359 00:15:45,240 --> 00:15:47,520 FIBROBLASTS AND THE CELLS ARE 360 00:15:47,520 --> 00:15:50,720 PROLIFERATING AND ISO GENIC TO 361 00:15:50,720 --> 00:15:51,480 THE STEM CELL. 362 00:15:51,480 --> 00:15:52,760 SO WE HAVE TWO POPULATION OF 363 00:15:52,760 --> 00:15:56,440 CELLS GROWING AND THE DIFFERENCE 364 00:15:56,440 --> 00:16:00,520 IS JUST DIFFERENTIATION STATE 365 00:16:00,520 --> 00:16:02,080 AND SHE COULD SEE WHAT HAPPENS. 366 00:16:02,080 --> 00:16:04,840 AND THE RESULT IS ONE OF THOSE 367 00:16:04,840 --> 00:16:07,400 RESULTS THAT TO ME I'VE SEEN 368 00:16:07,400 --> 00:16:09,320 MANY TIMES AND SPOKE ABOUT IT 369 00:16:09,320 --> 00:16:10,560 MANY TIMES AND ALL THE TIMES I 370 00:16:10,560 --> 00:16:13,280 SAY IT AND ALL THE TIME STILL 371 00:16:13,280 --> 00:16:14,480 SHOCKED IT'S ALL BLACK AND 372 00:16:14,480 --> 00:16:15,040 WHITE. 373 00:16:15,040 --> 00:16:16,520 YOU HAVE THE EXPECTED PHENOTYPE 374 00:16:16,520 --> 00:16:18,760 AND WE HAVE SOMATIC CELLS AND 375 00:16:18,760 --> 00:16:20,560 END TO END CHROMOSOMAL FUSION 376 00:16:20,560 --> 00:16:24,520 AND SEE THE SPAGHETTI LIKE 377 00:16:24,520 --> 00:16:27,400 STRUCTURE ARE FUSED TOGETHER BY 378 00:16:27,400 --> 00:16:28,560 THE END PATHWAY AND WHAT WE SEE 379 00:16:28,560 --> 00:16:30,280 IN EVERY CELL. 380 00:16:30,280 --> 00:16:33,000 WE DID IT IN HEPATOCYTES AND 381 00:16:33,000 --> 00:16:36,840 EVERY CELL WE STARTED UP TO NOW 382 00:16:36,840 --> 00:16:38,760 SHOWS THIS PHENOTYPE. 383 00:16:38,760 --> 00:16:41,840 HOWEVER THE EMBRYONIC STEM CELLS 384 00:16:41,840 --> 00:16:43,920 DO NOT SHOW -- THEY SHOW HARDLY 385 00:16:43,920 --> 00:16:45,960 ANY TYPE OF FUSIONS. 386 00:16:45,960 --> 00:16:48,880 MORE STRIKING, THEY KEEP 387 00:16:48,880 --> 00:16:49,280 PROLIFERATING. 388 00:16:49,280 --> 00:16:53,760 THEY PROLIFERATE A LITTLE BIT 389 00:16:53,760 --> 00:16:55,080 LESS THAN THE COUNTERPART BUT 390 00:16:55,080 --> 00:16:57,200 STILL PROLIFERATE. 391 00:16:57,200 --> 00:16:59,160 SO THIS WAS TELLING US THERE WAS 392 00:16:59,160 --> 00:17:01,400 SOMETHING UNIQUE ABOUT THESE 393 00:17:01,400 --> 00:17:03,720 CELLS WE COULD DELETE TR2 BUT 394 00:17:03,720 --> 00:17:05,800 SOMETHING WAS MODIFYING THE 395 00:17:05,800 --> 00:17:06,040 PHENOTYPE. 396 00:17:06,040 --> 00:17:07,040 THAT'S ONE OF THE THINGS WE 397 00:17:07,040 --> 00:17:08,360 WERE LOOKING FOR. 398 00:17:08,360 --> 00:17:09,440 THIS TRIGGERED A LOT OF WORK 399 00:17:09,440 --> 00:17:12,360 FROM OUR PART TO TRY AND 400 00:17:12,360 --> 00:17:16,520 UNDERSTAND WHAT IS THIS PATHWAY 401 00:17:16,520 --> 00:17:19,360 AND WHAT CAN PREVENT FULL BLOWN 402 00:17:19,360 --> 00:17:22,520 DNA DAMAGE RESPONSE OF 403 00:17:22,520 --> 00:17:23,640 CHROMOSOMAL ENDS. 404 00:17:23,640 --> 00:17:27,720 WE WERE NOT ALONE IN THIS. 405 00:17:27,720 --> 00:17:30,880 SIMON BOLTON'S LAB CAME TO A 406 00:17:30,880 --> 00:17:31,800 SIMILAR CONCLUSION AND PUBLISHED 407 00:17:31,800 --> 00:17:33,720 A PAPER A COUPLE YEARS AGO. 408 00:17:33,720 --> 00:17:35,200 AND I'M NOT GOING TO GO INTO ALL 409 00:17:35,200 --> 00:17:37,360 THE DETAILS OF WHAT WE PUBLISHED 410 00:17:37,360 --> 00:17:39,440 BUT LET ME TELL YOU WHEN WE 411 00:17:39,440 --> 00:17:41,880 PUBLISHED A COUPLE IDEAS THE WAY 412 00:17:41,880 --> 00:17:46,400 THIS COULD WORK SO ONE 413 00:17:46,400 --> 00:17:49,800 HYPOTHESIS THAT THE TELOMERES 414 00:17:49,800 --> 00:17:51,960 HAD A DAMPENED DNA DAMAGE 415 00:17:51,960 --> 00:17:53,320 RESPONSE AND FOUND MOLECULES 416 00:17:53,320 --> 00:17:55,200 THAT COULD TRIGGER THE RESPONSE 417 00:17:55,200 --> 00:17:57,160 AND THE MOST STRIKING, EXCITING 418 00:17:57,160 --> 00:17:58,680 IN MANY WAYS FOR ME WAS THE FACT 419 00:17:58,680 --> 00:18:03,240 WE SAW INDUCTION OF A SET OF 420 00:18:03,240 --> 00:18:04,600 GENES USUALLY EXPRESSED EARLY 421 00:18:04,600 --> 00:18:09,040 DURING EMBRYOGENESIS AND SEEM TO 422 00:18:09,040 --> 00:18:10,360 BE INVOLVED IN THE PROTECTION 423 00:18:10,360 --> 00:18:13,320 PHENOTYPE AND AMONG THOSE WERE A 424 00:18:13,320 --> 00:18:17,360 GENE WHICH I KNEW NOTHING ABOUT 425 00:18:17,360 --> 00:18:19,240 AT THAT POINT BUT SEEMED 426 00:18:19,240 --> 00:18:19,520 INTERESTING. 427 00:18:19,520 --> 00:18:22,280 WE HAD SOME INKLING BUT NOT 428 00:18:22,280 --> 00:18:23,520 COMPLETELY SOLID WHETHER THIS 429 00:18:23,520 --> 00:18:26,520 WAS PROVIDING PROTECTION. 430 00:18:26,520 --> 00:18:29,560 SO MARTA DECIDED THE ONLY WAY TO 431 00:18:29,560 --> 00:18:35,000 KNOW IF THAT WAS INDEED THE CASE 432 00:18:35,000 --> 00:18:36,760 WAS TO DELETE THIS AND THE 433 00:18:36,760 --> 00:18:40,520 PROBLEM IS AT LEAST IN MICE 434 00:18:40,520 --> 00:18:49,600 THERE ARE SIX GENES THAT ENCODE 435 00:18:49,600 --> 00:18:52,440 ZSCAN4 AND THEY'RE COMPETITIVE 436 00:18:52,440 --> 00:18:54,160 AND HAVE EXTRA GENES IN THE 437 00:18:54,160 --> 00:18:56,720 MIDDLE SO WE CAN'T JUST DEPLETE 438 00:18:56,720 --> 00:18:58,320 THE LOCUS. 439 00:18:58,320 --> 00:19:00,400 WHEN WE SAW IT I THOUGHT THERE'S 440 00:19:00,400 --> 00:19:03,240 NO WAY TO KNOCK IT OUT AND MARTA 441 00:19:03,240 --> 00:19:06,960 IS VERY GOOD AND ALSO EXTREMELY 442 00:19:06,960 --> 00:19:11,360 STUBBORN AND KNOCKED EVERY 443 00:19:11,360 --> 00:19:13,960 ISOFORM OF ZSCAN4 AND THIS 444 00:19:13,960 --> 00:19:15,840 DOESN'T DO JUSTICE TO THE AMOUNT 445 00:19:15,840 --> 00:19:18,480 OF WORK SHE DID BECAUSE IT TOOK 446 00:19:18,480 --> 00:19:20,640 A LONG TIME AND ONCE WE DID THIS 447 00:19:20,640 --> 00:19:24,480 WE HAVE A CLEAN -- TWO ACTUALLY 448 00:19:24,480 --> 00:19:28,520 CLEAN ZSCAN4 KNOCKOUTS SO THE 449 00:19:28,520 --> 00:19:30,600 RESULT WAS CLEAR. 450 00:19:30,600 --> 00:19:32,520 WHEN WE DELETE IT IN THE 451 00:19:32,520 --> 00:19:34,560 PRESENCE OF ZSCAN 4 WE HAVE 452 00:19:34,560 --> 00:19:35,840 LITTLE FUSIONS AND IN THE 453 00:19:35,840 --> 00:19:38,000 ABSENCE OF THIS WE START TO GET 454 00:19:38,000 --> 00:19:48,520 CHAINS OF CHROMOSOME FUSIONS. 455 00:19:50,040 --> 00:19:52,120 SO MARTA IS NOW STUDYING THE 456 00:19:52,120 --> 00:19:54,200 ZSCAN4 AND SHE IS NOW ON THE JOB 457 00:19:54,200 --> 00:19:57,360 MARKET SO LIKELY FIGURE OUT WHAT 458 00:19:57,360 --> 00:19:59,000 ZSCAN4 DOES IN HER OWN LAB WHICH 459 00:19:59,000 --> 00:20:03,360 IS VERY EXCITING BUT FROM 460 00:20:03,360 --> 00:20:04,320 ANOTHER POINT OF VIEW YOU CAN 461 00:20:04,320 --> 00:20:09,040 SEE ON THE ONE HAND IT'S GOOD WE 462 00:20:09,040 --> 00:20:13,360 CAN CONFIRM WHAT WE SUSPECTED SO 463 00:20:13,360 --> 00:20:14,560 ZSCAN4 IS DOING SOMETHING BUT 464 00:20:14,560 --> 00:20:16,000 NOT EVERYTHING BECAUSE WHAT I 465 00:20:16,000 --> 00:20:18,280 SHOWED IN THE SOMATIC CELL YOU 466 00:20:18,280 --> 00:20:19,760 GET THREE AND EVERY CHROMOSOME 467 00:20:19,760 --> 00:20:21,760 IS FUSED TOGETHER AND THIS IS 468 00:20:21,760 --> 00:20:23,240 STILL NOT A FULL BLOWN PHENOTYPE 469 00:20:23,240 --> 00:20:25,080 THAT WE SEE HERE. 470 00:20:25,080 --> 00:20:27,120 SO THERE MUST BE SOMETHING ELSE. 471 00:20:27,120 --> 00:20:30,080 SO MARTA THEN JOINED FORCES WITH 472 00:20:30,080 --> 00:20:34,600 ANOTHER POSTDOC IN THE LAB AND 473 00:20:34,600 --> 00:20:37,240 THEY DECIDED TO SEE IF THERE WAS 474 00:20:37,240 --> 00:20:39,320 ANYTHING WE WERE MISSING AND 475 00:20:39,320 --> 00:20:42,800 DOING GENETIC SCREENS USING 476 00:20:42,800 --> 00:20:45,440 CRISPR AND KNOCKOUT EVERY SINGLE 477 00:20:45,440 --> 00:20:49,560 GENE AND THEN ASK IF THERE IS 478 00:20:49,560 --> 00:20:51,760 ANY GENE THAT IS KILLING 479 00:20:51,760 --> 00:20:52,400 SELECTIVELY ZSCAN4 KNOCKOUT 480 00:20:52,400 --> 00:20:54,960 CELLS AND MAYBE THERE'S A 481 00:20:54,960 --> 00:20:57,600 PROTECTIVE GENE IN THE GENOME 482 00:20:57,600 --> 00:20:59,880 THAT IS PROTECTING TR2 KNOCKOUT 483 00:20:59,880 --> 00:21:03,560 CELLS AND WE THOUGHT THAT GENE 484 00:21:03,560 --> 00:21:06,600 WOULD DIE HOWEVER, IN TR2 485 00:21:06,600 --> 00:21:08,480 PROFICIENT CELLS IT WOULD NOT 486 00:21:08,480 --> 00:21:08,920 HAPPEN. 487 00:21:08,920 --> 00:21:10,920 SO WE DID THIS AND I'M GOING 488 00:21:10,920 --> 00:21:12,520 GIVE YOU JUST ONE IDEA OF THE 489 00:21:12,520 --> 00:21:14,040 WAY WE SEE THIS. 490 00:21:14,040 --> 00:21:15,880 SO USUALLY WHAT YOU DO IS YOU 491 00:21:15,880 --> 00:21:17,160 GET A SCORE. 492 00:21:17,160 --> 00:21:19,360 DEPENDING IF THE GENE IS 493 00:21:19,360 --> 00:21:22,280 ESSENTIAL IT'S GOING TO BE THOSE 494 00:21:22,280 --> 00:21:23,160 CELLS EXPRESSED IN THE RNA WOULD 495 00:21:23,160 --> 00:21:25,000 BE LOST FROM THE POPULATION. 496 00:21:25,000 --> 00:21:27,120 AND WE GIVE A BETA SCORE. 497 00:21:27,120 --> 00:21:30,160 SO THE MORE NEGATIVE VALUE YOU 498 00:21:30,160 --> 00:21:34,280 GET THE MORE ESSENTIAL A GENE IN 499 00:21:34,280 --> 00:21:34,680 THESE SITUATIONS. 500 00:21:34,680 --> 00:21:36,400 WHAT I'M SHOWING IS THE TOP 501 00:21:36,400 --> 00:21:37,440 REPRESENTATION OF THE OLD GENOME 502 00:21:37,440 --> 00:21:40,280 AND THE GENES THAT ARE DOWN HERE 503 00:21:40,280 --> 00:21:41,800 SELECTIVELY LINKED HERE TO 504 00:21:41,800 --> 00:21:42,440 KNOCKOUT CELLS. 505 00:21:42,440 --> 00:21:45,320 THERE'S A COUPLE OF GENES IN 506 00:21:45,320 --> 00:21:45,560 YELLOW. 507 00:21:45,560 --> 00:21:48,400 I WON'T GO INTO GENES WE 508 00:21:48,400 --> 00:21:49,080 VERIFIED PREVIOUSLY IN THE LAB. 509 00:21:49,080 --> 00:21:51,160 ONE THING THAT WAS STRIKING FOR 510 00:21:51,160 --> 00:21:53,840 US WE SAW A LOT OF GENES WITH 511 00:21:53,840 --> 00:21:54,760 THE SAME NAME. 512 00:21:54,760 --> 00:21:56,960 AND WHEN WE LOOKED FURTHER DOWN 513 00:21:56,960 --> 00:21:58,920 THERE WAS ADDITIONAL GENE ALL 514 00:21:58,920 --> 00:22:01,280 BELONGING TO THE SAME FAMILY. 515 00:22:01,280 --> 00:22:06,600 THE FAMILY KNOWN AND IT WAS 516 00:22:06,600 --> 00:22:07,360 SURPRISING FOR ME. 517 00:22:07,360 --> 00:22:11,080 IT'S A PATHWAY THAT IS WELL 518 00:22:11,080 --> 00:22:14,080 KNOWN TO BE SCANNING THE 519 00:22:14,080 --> 00:22:18,120 TRANSCRIPTOME AND REMOVING ANY 520 00:22:18,120 --> 00:22:19,120 GENE FOR INSTANCE. 521 00:22:19,120 --> 00:22:21,680 BASICALLY A QUALITY CONTROL FOR 522 00:22:21,680 --> 00:22:24,280 RNA. 523 00:22:24,280 --> 00:22:28,160 AND SO WHY WOULD THE PATHWAY BE 524 00:22:28,160 --> 00:22:31,320 LINKED WITH TR2 THAT WAS NOT 525 00:22:31,320 --> 00:22:32,600 IMMEDIATELY APPARENT AND THERE 526 00:22:32,600 --> 00:22:34,600 WAS SOME LITERATURE. 527 00:22:34,600 --> 00:22:39,160 SOME OF THE GENES SEEM TO BE 528 00:22:39,160 --> 00:22:43,360 CLOSE TO WHAT IS INVOLVED IN 529 00:22:43,360 --> 00:22:44,360 TELOMERASE ACTIVITY. 530 00:22:44,360 --> 00:22:45,440 WE THOUGHT MAYBE THERE'S 531 00:22:45,440 --> 00:22:46,080 SOMETHING ELSE. 532 00:22:46,080 --> 00:22:47,800 WE WANTED TO TEST IF THAT WAS 533 00:22:47,800 --> 00:22:49,240 THE CASE. 534 00:22:49,240 --> 00:22:51,920 WE WOULD DO THIS IN THE LAB AND 535 00:22:51,920 --> 00:22:54,520 WE DELETE THE GENES AND ASK IF 536 00:22:54,520 --> 00:22:56,960 THEY'RE VIABLE AND IF THEY'RE 537 00:22:56,960 --> 00:22:58,720 NOT CAN WE DO ANYTHING TO SEE 538 00:22:58,720 --> 00:23:00,240 WHETHER THEY PLAY A ROLE IN THE 539 00:23:00,240 --> 00:23:00,520 CELLS. 540 00:23:00,520 --> 00:23:01,880 IT WAS TOLD THE PATHWAY IS 541 00:23:01,880 --> 00:23:04,120 ESSENTIAL IN SOMATIC CELLS IS 542 00:23:04,120 --> 00:23:06,800 NOT ESSENTIAL IN THESE CELLS. 543 00:23:06,800 --> 00:23:07,840 WE WERE NOT THE FIRST TO OBSERVE 544 00:23:07,840 --> 00:23:10,280 THIS BUT CONFIRMED THIS. 545 00:23:10,280 --> 00:23:14,120 WE DELETED A LOT OF THE NMD 546 00:23:14,120 --> 00:23:15,600 FACTORS AND THEY CAN VERIFY THEY 547 00:23:15,600 --> 00:23:20,280 DID KNOCKOUTS IN DIFFERENT WAYS 548 00:23:20,280 --> 00:23:24,400 AND IMPORTANTLY THEN WE COULD 549 00:23:24,400 --> 00:23:25,560 SHOW THAT THEY INDEED INVOLVED 550 00:23:25,560 --> 00:23:26,560 IN THE PATHWAY. 551 00:23:26,560 --> 00:23:31,360 WE DID THIS THANKS WITH A LOT OF 552 00:23:31,360 --> 00:23:41,640 HELP FROM NIH. 553 00:23:45,840 --> 00:23:48,280 AND HERE WE'RE GIVING YOU A 554 00:23:48,280 --> 00:23:51,760 ROUGH OVERVIEW SO YOU TEND TO 555 00:23:51,760 --> 00:23:53,160 HAVE A SPLICE ISOFORM. 556 00:23:53,160 --> 00:23:56,240 FOR INSTANCE, YOU CAN HAVE CASES 557 00:23:56,240 --> 00:23:58,840 IN WHICH THEY'RE RETAINED OR 558 00:23:58,840 --> 00:24:02,040 CASES IN WHICH THEY'RE SKIPPED. 559 00:24:02,040 --> 00:24:04,520 THOSE TYPE OF TRANSCRIPTS ARE 560 00:24:04,520 --> 00:24:09,720 NORMALLY DEPLETED BY THE PATHWAY 561 00:24:09,720 --> 00:24:12,200 BUT IN THE ABSENCE OF THESE 562 00:24:12,200 --> 00:24:14,360 GENES THE EVENTS ARE ACCUMULATED 563 00:24:14,360 --> 00:24:16,520 AND WE SEE A LOT OF THEM 564 00:24:16,520 --> 00:24:20,640 CONFIRMING THE PROTEINS DO PLAY 565 00:24:20,640 --> 00:24:22,280 A ROLE IN THE PATHWAY IN STEM 566 00:24:22,280 --> 00:24:29,560 CELLS. 567 00:24:29,560 --> 00:24:30,160 AND THE RESULT WAS REALLY 568 00:24:30,160 --> 00:24:35,200 STRIKING. 569 00:24:35,200 --> 00:24:38,720 SO HERE IS THE KNOCKOUT BY 570 00:24:38,720 --> 00:24:39,000 THEMSELVES. 571 00:24:39,000 --> 00:24:40,840 THE CHROMOSOMES ARE REALLY NICE 572 00:24:40,840 --> 00:24:42,000 FROM WHAT WE CAN TELL. 573 00:24:42,000 --> 00:24:46,440 THERE'S NO FUSIONS THEY LOOK 574 00:24:46,440 --> 00:24:48,240 PRETTY HOWEVER, WHEN WE DELETE 575 00:24:48,240 --> 00:24:51,040 TR2 IN THE CONTEXT WE CAN SEE 576 00:24:51,040 --> 00:24:54,840 THE ENGAGEMENT OF END 577 00:24:54,840 --> 00:24:59,320 CHROMESOMAL FUSION AND THIS IS 578 00:24:59,320 --> 00:25:01,360 TRUE FOR THE MEMBERS OF THE 579 00:25:01,360 --> 00:25:03,360 PATHWAY WE TEFFED TODAY. 580 00:25:03,360 --> 00:25:04,680 -- TESTED TODAY. 581 00:25:04,680 --> 00:25:07,280 THERE'S A SLIGHT VARIATION WE 582 00:25:07,280 --> 00:25:08,520 STILL DON'T UNDERSTAND FROM IT'S 583 00:25:08,520 --> 00:25:12,240 TRUE BIOLOGICAL OR NOT BUT MANY 584 00:25:12,240 --> 00:25:13,800 HAVE HIGH LEVEL OF CHROMOSOMAL 585 00:25:13,800 --> 00:25:14,160 FUSIONS. 586 00:25:14,160 --> 00:25:19,480 AND TO PUT IT IN CONTEXT THIS IS 587 00:25:19,480 --> 00:25:22,040 WHAT WE SAW WITH WHAT WE SEE IN 588 00:25:22,040 --> 00:25:23,240 SOMATIC LEVELS AROUND 50% AND 589 00:25:23,240 --> 00:25:25,480 HERE WE GET 20% OF FUSION. 590 00:25:25,480 --> 00:25:28,920 WE ARE STILL NOT AT THE LEVEL OF 591 00:25:28,920 --> 00:25:29,880 SOMATIC CELLS BUT IT'S A LOT OF 592 00:25:29,880 --> 00:25:40,120 TELEFUSIONS. 593 00:25:53,240 --> 00:25:55,360 WE WANT TO SEE IF THERE'S A SET 594 00:25:55,360 --> 00:25:57,640 OF CELLS THAT CAN PROTECT 595 00:25:57,640 --> 00:26:01,200 TELOMERES IN THE ABSENCE OF TR2. 596 00:26:01,200 --> 00:26:05,720 THE GENES IN THE ABSENCE OF NMD 597 00:26:05,720 --> 00:26:07,640 PATHWAY ARE EXPRESSED AT LOWER 598 00:26:07,640 --> 00:26:07,840 LEVEL. 599 00:26:07,840 --> 00:26:11,800 WE'RE GETTING THERE AND WHAT I 600 00:26:11,800 --> 00:26:16,560 MEAN IS WE DID TRANSCRIPTIONAL 601 00:26:16,560 --> 00:26:19,520 PROFILING AND FOUND AROUND 100 602 00:26:19,520 --> 00:26:22,720 GENES THAT ARE CONSISTENTLY 603 00:26:22,720 --> 00:26:24,520 AFFECTED IN NMD PATHWAY AND 604 00:26:24,520 --> 00:26:25,680 THINK SOME OF THOSE ARE THE ONES 605 00:26:25,680 --> 00:26:28,000 AND WE STILL HAVE TO DO A NUMBER 606 00:26:28,000 --> 00:26:35,760 OF THINGS TO VERIFY THIS. 607 00:26:35,760 --> 00:26:37,080 AND HOPEFULLY WE'LL HAVE AN 608 00:26:37,080 --> 00:26:37,360 UPDATE SOON. 609 00:26:37,360 --> 00:26:43,280 OKAY. 610 00:26:43,280 --> 00:26:46,320 AND THIS CONCLUDES THE PART ON 611 00:26:46,320 --> 00:26:47,520 THE TELOMERE PROTECTION AND 612 00:26:47,520 --> 00:26:49,040 GOING BACK TO THE ORIGINAL 613 00:26:49,040 --> 00:26:50,960 QUESTION DOES IT TRANSLATE TO 614 00:26:50,960 --> 00:26:52,000 ANYTHING IMPORTANT POTENTIALLY 615 00:26:52,000 --> 00:26:53,560 FOR CANCER AND I THINK SO. 616 00:26:53,560 --> 00:26:55,680 WE LIKE TO THINK SO. 617 00:26:55,680 --> 00:26:58,400 WE WERE LOOKING FOR FACTORS THAT 618 00:26:58,400 --> 00:27:00,320 COULD MEDIATE THE RESPONSE TO 619 00:27:00,320 --> 00:27:08,920 VERY SHORT TELOMERES. 620 00:27:08,920 --> 00:27:10,680 INTERESTINGLY THERE'S A GENE NOT 621 00:27:10,680 --> 00:27:17,120 EXPRESSED IN SOMATIC CELLS ONLY 622 00:27:17,120 --> 00:27:19,960 EARLY DURING EMBRYOGENESIS AND 623 00:27:19,960 --> 00:27:23,760 YOU HAVE SOME THAT DO EXPRESS 624 00:27:23,760 --> 00:27:25,240 ZSCAN4 SO IT'S TEMPTING TO 625 00:27:25,240 --> 00:27:27,120 SPECULATE AND MAYBE IN THOSE 626 00:27:27,120 --> 00:27:30,840 CELLS THE INDUCTION COULD HAVE 627 00:27:30,840 --> 00:27:32,520 HELPED SOMEHOW TELOMERE EROSION. 628 00:27:32,520 --> 00:27:38,360 THIS IS SOMETHING MARTA IS 629 00:27:38,360 --> 00:27:48,560 WISHING TO TEST IN HER OWN 630 00:27:48,560 --> 00:27:52,840 RESEARCH AND NMD HAS BEEN SHOWN 631 00:27:52,840 --> 00:27:54,840 TO BE POTENTIALLY IN SOME 632 00:27:54,840 --> 00:27:55,880 SETTINGS FOR INSTANCE IN CELLS 633 00:27:55,880 --> 00:27:58,200 WITH THE ABILITY THAT HAVE A LOT 634 00:27:58,200 --> 00:28:00,160 OF INSTABILITY SO POTENTIALLY 635 00:28:00,160 --> 00:28:02,000 THERE COULD BE A LINK WITH 636 00:28:02,000 --> 00:28:02,800 TELOMERE DYSFUNCTION AS WELL AND 637 00:28:02,800 --> 00:28:06,760 THIS IS SOMETHING WE NEED TO 638 00:28:06,760 --> 00:28:08,000 TEST. 639 00:28:08,000 --> 00:28:08,440 OKAY. 640 00:28:08,440 --> 00:28:12,440 SO FOR THE LAST PART OF MY TALK 641 00:28:12,440 --> 00:28:16,480 I'M GOING TO INSTEAD TELL YOU 642 00:28:16,480 --> 00:28:20,080 SOMETHING ABOUT THE MECHANISM OF 643 00:28:20,080 --> 00:28:22,400 TELOMERE ELONGATION. 644 00:28:22,400 --> 00:28:32,920 IT'S AN AREA THAT IS DEF AND THE 645 00:28:32,920 --> 00:28:38,600 IDEA IS WHEN CELLS UNDER GO 646 00:28:38,600 --> 00:28:40,880 MASSIVE TELEMERE EROSION THEY 647 00:28:40,880 --> 00:28:44,800 CAN ELONGATE AND ALLOW THEM TO 648 00:28:44,800 --> 00:28:46,480 BE IMMORTAL. 649 00:28:46,480 --> 00:28:51,480 HOW'S THIS WORK. 650 00:28:51,480 --> 00:28:55,920 ROUGHLY 85% OF HUMAN CANCER IS 651 00:28:55,920 --> 00:28:59,760 EXPRESSED IN TELOMERASE AND 652 00:28:59,760 --> 00:29:06,560 THERE'S PROMOTER IN THE 653 00:29:06,560 --> 00:29:08,320 TELOMERASE THAT ALLOWS 654 00:29:08,320 --> 00:29:13,720 EXPRESSION BUT HOW'S IT 655 00:29:13,720 --> 00:29:21,520 FUNCTION? 656 00:29:21,520 --> 00:29:24,800 WHOEN YOU THINK ABOUT IT 657 00:29:24,800 --> 00:29:28,360 TELOMERASE IS A TRICKY ENZYME TO 658 00:29:28,360 --> 00:29:28,640 FUNCTION. 659 00:29:28,640 --> 00:29:31,600 THE BRIAN LAB ESTIMATED ROUGHLY 660 00:29:31,600 --> 00:29:36,400 10 TO 15 MOLECULES OF TELOMERASE 661 00:29:36,400 --> 00:29:39,120 AND IN CANCER CELLS THAT 662 00:29:39,120 --> 00:29:41,080 OVEREXPRESS TELOMERASE SO IT'S 663 00:29:41,080 --> 00:29:45,800 AN ABUNDANT MOLECULE AND THERE'S 664 00:29:45,800 --> 00:29:49,960 A LOT OF GENOME TO SCAN. 665 00:29:49,960 --> 00:29:54,560 SO HOW'S IT FIND THE END TO ACT 666 00:29:54,560 --> 00:29:55,320 IS UNCLEAR. 667 00:29:55,320 --> 00:29:57,160 ONE INDICATION OF HOW IT COULD 668 00:29:57,160 --> 00:30:00,440 WORK CAME OUT THANKS TO THE WORK 669 00:30:00,440 --> 00:30:05,240 OF TOM CHECK WHICH IDENTIFIED AN 670 00:30:05,240 --> 00:30:06,920 INTERACTION BETWEEN TELOMERASE 671 00:30:06,920 --> 00:30:12,360 AND THE COMPLEX I TOLD YOU 672 00:30:12,360 --> 00:30:15,280 BEFORE, AND THIS ALLOWS THEM TO 673 00:30:15,280 --> 00:30:18,600 FIND TELOMERES BUT IT'S NOT 674 00:30:18,600 --> 00:30:18,920 SUFFICIENT. 675 00:30:18,920 --> 00:30:21,400 WHILE I'M DRAWING THIS IN A 676 00:30:21,400 --> 00:30:24,280 SIMPLIFIED MANNER WITH THE END 677 00:30:24,280 --> 00:30:26,160 OF THE CHROMOSOME ENDS WITH THIS 678 00:30:26,160 --> 00:30:28,040 PROTEIN COMPLEX HERE, THE 679 00:30:28,040 --> 00:30:29,680 REALITY IS A LOT DIFFERENT. 680 00:30:29,680 --> 00:30:32,480 IN REALITY YOU CAN LOOK AT A 681 00:30:32,480 --> 00:30:33,960 SINGLE CHROMOSOME AND THE 682 00:30:33,960 --> 00:30:34,680 PROTEIN COMPLEX IS SPREAD 683 00:30:34,680 --> 00:30:37,120 THROUGHOUT THE ENTIRETY OF THE 684 00:30:37,120 --> 00:30:38,920 CHROMOSOME OR TELOMERE. 685 00:30:38,920 --> 00:30:43,360 SO THIS INTERACTION CAN LOCALIZE 686 00:30:43,360 --> 00:30:44,640 TELOMERASE TO THE TELOMERES BUT 687 00:30:44,640 --> 00:30:46,280 ALSO POTENTIALLY A PROBLEM 688 00:30:46,280 --> 00:30:50,040 BECAUSE THEN YOU'LL HAVE 689 00:30:50,040 --> 00:30:51,440 UNPRODUCTIVE INTERACTION ALONG 690 00:30:51,440 --> 00:30:52,400 THE TELOMERE. 691 00:30:52,400 --> 00:30:54,280 HOW CAN TELOMERASE BE TREATED 692 00:30:54,280 --> 00:30:54,560 HERE. 693 00:30:54,560 --> 00:30:56,840 THIS IS ONE WAY OF SAYING WE 694 00:30:56,840 --> 00:30:57,280 DON'T KNOW. 695 00:30:57,280 --> 00:30:59,120 SO THERE'S SOMETHING ELSE WE'RE 696 00:30:59,120 --> 00:31:00,360 MISSING. 697 00:31:00,360 --> 00:31:02,920 SOMETHING THAT NEEDS TO BE FOUND 698 00:31:02,920 --> 00:31:06,560 THAT CAN EXPLAIN HOW TELOMERASE 699 00:31:06,560 --> 00:31:12,480 CAN BE AT THE END IN OUR 700 00:31:12,480 --> 00:31:12,720 MODELLING. 701 00:31:12,720 --> 00:31:15,440 SO THE NEWEST MEMBER OF THE LAB 702 00:31:15,440 --> 00:31:19,200 JOINED A YEAR AND A HALF AGO AND 703 00:31:19,200 --> 00:31:22,320 HE'S BEEN A PHENOMENAL IN THE 704 00:31:22,320 --> 00:31:24,800 LAB IN LITTLE TIME MANAGED TO 705 00:31:24,800 --> 00:31:28,520 GENERATE A LOT OF DATA AND 706 00:31:28,520 --> 00:31:32,760 REALLY COOL THINGS TO STUDY 707 00:31:32,760 --> 00:31:34,120 TELOMERASE ACTIVITY AND FIRST 708 00:31:34,120 --> 00:31:35,720 IDENTIFIED ANY POTENTIAL 709 00:31:35,720 --> 00:31:37,480 CANDIDATE THAT COULD INTERACT 710 00:31:37,480 --> 00:31:41,520 WITH TELOMERASE IN TWO WAYS. 711 00:31:41,520 --> 00:31:48,040 THE FIRST WAY WAS A PROTEOMICS 712 00:31:48,040 --> 00:31:49,840 BASED APPROACHED AND USED 713 00:31:49,840 --> 00:31:54,080 PROXIMITY BASED LIGATION AND 714 00:31:54,080 --> 00:31:55,800 PROTEINS THAT COME IN CLOSE 715 00:31:55,800 --> 00:31:57,800 PROXIMITY TO A BAIT AND USE THE 716 00:31:57,800 --> 00:32:03,560 PROTEIN KNOWN TO INTERACT WITH 717 00:32:03,560 --> 00:32:07,360 TELOMERASE AND THERE'S A MUTANT 718 00:32:07,360 --> 00:32:09,640 THAT CANNOT ENACT AND COMPARING 719 00:32:09,640 --> 00:32:12,360 THE TWO SITUATIONS HE'S HOPING 720 00:32:12,360 --> 00:32:14,960 TO FIND FACTORS BOUND IN 721 00:32:14,960 --> 00:32:19,120 PROXIMITY TO TELOMERASE AND 722 00:32:19,120 --> 00:32:19,920 POTENTIALLY TO TELOMERASE AND 723 00:32:19,920 --> 00:32:23,360 EXCLUDE THE ONE THAT INTERACTS 724 00:32:23,360 --> 00:32:25,920 WITH MPP1. 725 00:32:25,920 --> 00:32:30,200 THERE'S THE KNOWN COMPONENT OF 726 00:32:30,200 --> 00:32:32,920 TELOMERASE AND HAS WHAT HAPPENS 727 00:32:32,920 --> 00:32:39,360 WHEN HE DOES THIS DONE IN 728 00:32:39,360 --> 00:32:42,240 COLLABORATION WITH THE 729 00:32:42,240 --> 00:32:44,880 SPECTROMETRY CORPS. 730 00:32:44,880 --> 00:32:46,400 YOU HAVE A LOT OF POTENTIAL HITS 731 00:32:46,400 --> 00:32:49,640 THIS WAY AND IT CAN BE DIFFICULT 732 00:32:49,640 --> 00:32:51,240 TO SEE EACH OF THEM IF YOU WANT 733 00:32:51,240 --> 00:32:53,960 TO GO AFTER EACH OF THEM. 734 00:32:53,960 --> 00:32:55,400 INSTEAD HE DID TWO EXTRA STEPS 735 00:32:55,400 --> 00:32:56,840 TO CLEAN UP THE LIST. 736 00:32:56,840 --> 00:33:01,720 THE FIRST WAS TO DO A CRISPR 737 00:33:01,720 --> 00:33:04,200 SCREEN AND FIND ANYTHING IT THAT 738 00:33:04,200 --> 00:33:05,920 IS ESSENTIAL FOR TELOMERASE 739 00:33:05,920 --> 00:33:08,720 ACTIVITY AND HE HAS PARENTAL 740 00:33:08,720 --> 00:33:11,240 CELL LINES WITH REALLY LONG 741 00:33:11,240 --> 00:33:14,640 TELOMERES AND CREATED CELL LINES 742 00:33:14,640 --> 00:33:18,160 THAT SURVIVE THANKS TO THE 743 00:33:18,160 --> 00:33:20,080 TELOMERASE ACTIVITY WITH SHORT 744 00:33:20,080 --> 00:33:20,360 TELOMERES. 745 00:33:20,360 --> 00:33:22,880 THESE ARE THE LIMITATION THAT 746 00:33:22,880 --> 00:33:25,360 ALLOWS THEM TO SURVIVE BUT JUST 747 00:33:25,360 --> 00:33:28,520 BARELY HANGING ON THANKS TO 748 00:33:28,520 --> 00:33:29,520 TELOMERASE ACTIVITY. 749 00:33:29,520 --> 00:33:31,760 SO THEN THE TWO SITUATIONS 750 00:33:31,760 --> 00:33:32,760 SHOULD BE GENETICALLY IDENTICAL 751 00:33:32,760 --> 00:33:35,440 AND NOW WE CAN DO A CRISPR 752 00:33:35,440 --> 00:33:36,880 SCREEN WHERE ASK WHO IS 753 00:33:36,880 --> 00:33:39,120 RESPONSIBLE FOR THE VIABILITY OF 754 00:33:39,120 --> 00:33:41,600 THE CELLS WITH SHORT TELOMERES. 755 00:33:41,600 --> 00:33:43,400 AND WHEN HE DID THIS HE FOUND A 756 00:33:43,400 --> 00:33:48,480 LOT OF GENES AND EXCITINGLY HE 757 00:33:48,480 --> 00:33:50,680 FOUND TELOMERASE SUGGESTING 758 00:33:50,680 --> 00:33:52,280 WE'RE HITTING THE PATHWAY WE 759 00:33:52,280 --> 00:33:53,600 THINK. 760 00:33:53,600 --> 00:33:55,240 AGAIN, THE TWO THINGS COMBINED 761 00:33:55,240 --> 00:33:58,880 IS STILL HUNDREDS OF GENES 762 00:33:58,880 --> 00:34:00,640 DIFFICULT TO ANALYZE ESPECIALLY 763 00:34:00,640 --> 00:34:03,400 BECAUSE THE ONLY WAY WE REALLY 764 00:34:03,400 --> 00:34:04,640 HAVE TO DATE TO MEASURE 765 00:34:04,640 --> 00:34:06,800 TELOMERASE ACTIVITY IS TO DO 766 00:34:06,800 --> 00:34:08,840 THESE TYPES OF EXPERIMENTS WHERE 767 00:34:08,840 --> 00:34:10,560 YOU TAKE CELLS, DELETE WHATEVER 768 00:34:10,560 --> 00:34:13,200 GENE YOU WANT TO AND CULTURE THE 769 00:34:13,200 --> 00:34:14,640 CELL. 770 00:34:14,640 --> 00:34:21,320 SO IN DOING THIS FOR 150 TARGET 771 00:34:21,320 --> 00:34:22,720 GENES IS DEMANDING AND HE WAS 772 00:34:22,720 --> 00:34:24,080 NOT GOING TO DO IT. 773 00:34:24,080 --> 00:34:25,480 INSTEAD WE THOUGHT OF TRYING TO 774 00:34:25,480 --> 00:34:27,920 DO A BETTER WAY TO TRY TO SEE IF 775 00:34:27,920 --> 00:34:30,680 WE COULD DEVELOP A WAY TO SEE 776 00:34:30,680 --> 00:34:32,200 TELOMERASE ACTIVITY IN A SINGLE 777 00:34:32,200 --> 00:34:34,520 CELL CYCLE THAT WOULD ALLOW TO 778 00:34:34,520 --> 00:34:35,520 US SCREEN ALL THE POTENTIAL 779 00:34:35,520 --> 00:34:43,600 TARGETS IN A RELIABLE AND FAST 780 00:34:43,600 --> 00:34:45,880 MAN 781 00:34:45,880 --> 00:34:47,680 MANNER IDEALLY WE'D LIKE TO SEE 782 00:34:47,680 --> 00:34:49,640 IT IN ACTION AND IT COMES WITH 783 00:34:49,640 --> 00:34:51,160 ITS OWN TEMPLATE AND USE IT TO 784 00:34:51,160 --> 00:35:00,280 ADD REPEATS AT THE END. 785 00:35:00,280 --> 00:35:02,960 IF WE COULD JUST HAVE REPEATS WE 786 00:35:02,960 --> 00:35:08,440 COULD DO IT BUT THEY'RE 787 00:35:08,440 --> 00:35:13,240 IDENTICAL TO THE REST. 788 00:35:13,240 --> 00:35:15,360 INSTEAD HE PROVIDED A MUTANT RNA 789 00:35:15,360 --> 00:35:18,480 AND CHANGE IT AT SEQUENCE AND HE 790 00:35:18,480 --> 00:35:23,680 CAN DETECT IT NICELY WITH A 791 00:35:23,680 --> 00:35:34,240 PROBE I WORKED NICELY BUT WHEN 792 00:35:36,280 --> 00:35:37,920 YOU INTRODUCE THE MUTATION IT 793 00:35:37,920 --> 00:35:40,920 DOESN'T ALLOW ALL THE PROTECTIVE 794 00:35:40,920 --> 00:35:46,000 PROTEINS TO BIND THEREFORE THE 795 00:35:46,000 --> 00:35:51,640 CELLS DIE. 796 00:35:51,640 --> 00:35:54,040 AS A RESULT IT DOESN'T REALLY 797 00:35:54,040 --> 00:35:55,680 WORK BUT IN THESE CELLS WE CAN 798 00:35:55,680 --> 00:35:57,040 GET RID OF THE PROTECTIVE 799 00:35:57,040 --> 00:35:59,160 PROTEINS AND IT SEEMS TO WORK. 800 00:35:59,160 --> 00:36:01,520 HE PUT THEM TOGETHER AND THOUGHT 801 00:36:01,520 --> 00:36:03,480 MAYBE WE CAN OVEREXPRESS THE 802 00:36:03,480 --> 00:36:05,080 MUTANT RNA IN THE CELLS AND SEE 803 00:36:05,080 --> 00:36:15,600 IF THIS CAN BE USED YOU HAVE 804 00:36:19,160 --> 00:36:22,200 CELLS THAT EXPRESS THE 805 00:36:22,200 --> 00:36:26,760 TELOMERASE AND WILD TYPE AND 806 00:36:26,760 --> 00:36:37,280 SOME WILL BE SYNTHESIZED AND YOU 807 00:36:38,720 --> 00:36:41,440 CAN SEE WILD TYPE AS WELL AS 808 00:36:41,440 --> 00:36:43,600 MUTANT ALMOST AT EVERY 809 00:36:43,600 --> 00:36:43,920 CHROMOSOME. 810 00:36:43,920 --> 00:36:45,880 I'M SAYING ALMOST BECAUSE SOME 811 00:36:45,880 --> 00:36:48,520 ENDS ARE NOT LABELLED. 812 00:36:48,520 --> 00:36:50,960 THAT MEANS TELOMERASE IN THE 813 00:36:50,960 --> 00:36:56,680 GIVEN DIVISIONS ACT IN THIS 814 00:36:56,680 --> 00:37:00,080 PARTICULAR TELOMERE. 815 00:37:00,080 --> 00:37:02,880 THE CELLS ARE VIABLE AND GROW 816 00:37:02,880 --> 00:37:05,840 AND SEEM TO BE COMPLETELY FINE. 817 00:37:05,840 --> 00:37:08,600 THIS MEANS WE HAVE A POTENTIAL 818 00:37:08,600 --> 00:37:10,080 ASSAY TO MEASURE TELOMERASE 819 00:37:10,080 --> 00:37:12,240 ACTIVITY IN A SINGLE CELL CYCLE 820 00:37:12,240 --> 00:37:14,680 OR AT LEAST IN THIS CYCLE. 821 00:37:14,680 --> 00:37:15,680 BUT THERE'S A FEW CONTROLS WE 822 00:37:15,680 --> 00:37:17,000 HAD TO DO. 823 00:37:17,000 --> 00:37:24,520 THE FIRST IS IS THIS REALLY 824 00:37:24,520 --> 00:37:28,080 TELOMERASE WE COULD TEST THIS 825 00:37:28,080 --> 00:37:29,320 QUITE EASILY AND DELETED 826 00:37:29,320 --> 00:37:34,200 TELOMERASE AND YOU CAN SEE IF 827 00:37:34,200 --> 00:37:36,600 YOU EXPRESS THE MUTANT 828 00:37:36,600 --> 00:37:38,000 TELOMERASE TEMPLATE IN WILD TYPE 829 00:37:38,000 --> 00:37:39,840 YOU CAN SEE THE MUTANT TEMPLATE 830 00:37:39,840 --> 00:37:42,040 AND THIS IS AN INTERFACE CELL 831 00:37:42,040 --> 00:37:44,200 AND WORKS NICELY IN INTERFACE 832 00:37:44,200 --> 00:37:48,840 CELLS BUT CANNOT GET COOPERATION 833 00:37:48,840 --> 00:37:51,960 IN TELOMERASE KNOCKOUT TELLING 834 00:37:51,960 --> 00:37:53,760 US YOU NEED A TO INCORPORATE 835 00:37:53,760 --> 00:37:55,360 THIS REPEAT. 836 00:37:55,360 --> 00:38:01,640 THAT'S GOOD. 837 00:38:01,640 --> 00:38:03,840 COULD IT BE TELOMERASE ACTIVITY 838 00:38:03,840 --> 00:38:05,560 IS DIFFERENT FROM EVERYTHING WE 839 00:38:05,560 --> 00:38:07,800 KNEW AND THEREFORE IS NOT REALLY 840 00:38:07,800 --> 00:38:10,080 RELEVANT FOR WHAT WE WANT TO 841 00:38:10,080 --> 00:38:10,360 STUDY. 842 00:38:10,360 --> 00:38:12,520 IN OTHER WORDS, CAN WE TEST 843 00:38:12,520 --> 00:38:12,800 ANYTHING? 844 00:38:12,800 --> 00:38:17,320 CAN WE DEPLETE ANYTHING TO HAVE 845 00:38:17,320 --> 00:38:19,520 AN EFFECT ON TELOMERASE IN HUMAN 846 00:38:19,520 --> 00:38:20,920 CANCER CELL LINES IS IMPORTANT. 847 00:38:20,920 --> 00:38:23,840 FOR THIS WHAT WE DID WAS TO 848 00:38:23,840 --> 00:38:29,520 DELETE TPP1 THE PROTEIN THAT 849 00:38:29,520 --> 00:38:33,840 BINDS TO TELOMERES AND SHOWN TO 850 00:38:33,840 --> 00:38:35,120 BE ESSENTIAL FOR TELOMERASE 851 00:38:35,120 --> 00:38:36,800 ACTIVITY AND IT WORKS NICELY. 852 00:38:36,800 --> 00:38:39,480 IF YOU DON'T HAVE TPP1 WE DON'T 853 00:38:39,480 --> 00:38:43,720 HAVE INCORPORATION OF THE MUTANT 854 00:38:43,720 --> 00:38:45,440 REPEAT AND IN THE WILD TYPE WE 855 00:38:45,440 --> 00:38:50,880 SEE IT AND JUST A QUANTIFICATION 856 00:38:50,880 --> 00:38:54,480 IF YOU DELETE A SECOND PROTEIN 857 00:38:54,480 --> 00:38:59,320 BOUND TO TELOMERES IT SHOWS IT'S 858 00:38:59,320 --> 00:39:01,840 NOT DELETING AS AN EFFECT AND 859 00:39:01,840 --> 00:39:04,080 NOW WE'RE IN BUSINESS AND HE CAN 860 00:39:04,080 --> 00:39:07,080 TAKE ALL THE CANDIDATE GENES AND 861 00:39:07,080 --> 00:39:09,480 PUT THEM IN HIGH THROUGHPUT 862 00:39:09,480 --> 00:39:10,800 FORMAT IN A SINGLE CELL CYCLE 863 00:39:10,800 --> 00:39:13,880 WHETHER OR NOT THEY HAVE A ROLE 864 00:39:13,880 --> 00:39:16,400 IN TELOMERE ELONGATION. 865 00:39:16,400 --> 00:39:20,480 IT DOES THIS USING AN INDUCIBLE 866 00:39:20,480 --> 00:39:22,480 SYSTEM CREATED INDUCED 867 00:39:22,480 --> 00:39:27,720 EXPRESSION OF THE MUTANT 868 00:39:27,720 --> 00:39:33,400 TELOMERASE AND LOOK DAYS AFTER 869 00:39:33,400 --> 00:39:35,600 FOR INCORPORATION. 870 00:39:35,600 --> 00:39:36,640 I THINK IT WILL A POWERFUL TOOL. 871 00:39:36,640 --> 00:39:37,880 IT'S ONGOING. 872 00:39:37,880 --> 00:39:39,520 I DON'T HAVE THE ANSWER BUT I 873 00:39:39,520 --> 00:39:40,760 THINK IT WOULD BE INTERESTING 874 00:39:40,760 --> 00:39:42,200 FOR A LOT OF YOU. 875 00:39:42,200 --> 00:39:47,640 ANYBODY THAT IS INTERESTED IN 876 00:39:47,640 --> 00:39:50,160 TELOMERASE ACTIVITY PLEASE COME 877 00:39:50,160 --> 00:39:54,080 AND TALK TO ME BECAUSE IT CAN 878 00:39:54,080 --> 00:39:54,720 SCREEN AND WE'RE EXCITED ABOUT 879 00:39:54,720 --> 00:40:00,680 THIS. 880 00:40:00,680 --> 00:40:01,680 HOPEFULLY WHEN WE COMPLETE THIS 881 00:40:01,680 --> 00:40:03,800 WE'LL HAVE A BETTER IDEA HOW 882 00:40:03,800 --> 00:40:06,320 THIS WORKS AND WHAT ARE FACTORS 883 00:40:06,320 --> 00:40:09,360 THAT CAN RECRUIT THEM TO 884 00:40:09,360 --> 00:40:09,960 TELOMERASE ENDS. 885 00:40:09,960 --> 00:40:13,160 THE LAST THING I'LL TELL YOU 886 00:40:13,160 --> 00:40:15,560 ABOUT IS ABOUT SOMETHING WE'VE 887 00:40:15,560 --> 00:40:17,960 BEEN WORKING QUITE A LOT OF WITH 888 00:40:17,960 --> 00:40:23,960 AND NOT TELL EVERYTHING AND THE 889 00:40:23,960 --> 00:40:26,720 KEY IS UPREGULATED IN WAYS WE 890 00:40:26,720 --> 00:40:28,400 DON'T UNDERSTAND IN ROUGHLY 50% 891 00:40:28,400 --> 00:40:30,800 OF TUMORS. 892 00:40:30,800 --> 00:40:31,880 GENETICALLY THERE MUST BE 893 00:40:31,880 --> 00:40:33,040 SOMETHING THAT TRIGGERS THE 894 00:40:33,040 --> 00:40:33,280 PATHWAY. 895 00:40:33,280 --> 00:40:38,000 SO WE KNOW THAT GENETICALLY 896 00:40:38,000 --> 00:40:41,320 USUALLY L POSITIVE CELLS HAVE 897 00:40:41,320 --> 00:40:43,480 EXPRESSION AND IT'S A CHAPERONE 898 00:40:43,480 --> 00:40:46,160 PROTEIN AND THE MUTATION ARE NOT 899 00:40:46,160 --> 00:40:48,680 SUFFICIENT TO TRIGGER SO THERE'S 900 00:40:48,680 --> 00:40:50,400 WE DON'T UNDERSTAND ABOUT THE 901 00:40:50,400 --> 00:40:50,680 BIOLOGY. 902 00:40:50,680 --> 00:40:53,080 BUT I THINK FROM A MORE 903 00:40:53,080 --> 00:40:53,760 PRACTICAL POINT OF VIEW WOULD BE 904 00:40:53,760 --> 00:40:56,560 COOL TO BE ABLE TO TARGET THIS 905 00:40:56,560 --> 00:40:57,040 PATHWAY. 906 00:40:57,040 --> 00:41:02,960 SO WE HAVE REALLY NO WAY OF 907 00:41:02,960 --> 00:41:04,400 TARGETING THE PATHWAY SO IF 908 00:41:04,400 --> 00:41:07,680 THERE'S A UNIQUE GENETIC 909 00:41:07,680 --> 00:41:09,120 SENSIBILITY IN THESE KIND OF 910 00:41:09,120 --> 00:41:10,400 CELLS WE DON'T KNOW AND ONE 911 00:41:10,400 --> 00:41:12,960 REASON WE DON'T KNOW IS WE 912 00:41:12,960 --> 00:41:16,040 REALLY STILL DON'T UNDERSTAND 913 00:41:16,040 --> 00:41:18,920 FULLY THE PATHWAY OF TELOMERE 914 00:41:18,920 --> 00:41:20,720 ELONGATION IN THE CELLS. 915 00:41:20,720 --> 00:41:23,000 HERE COMES A Ph.D. STUDENT IN 916 00:41:23,000 --> 00:41:27,760 THE LAB THAT JOINED ROUGHLY A 917 00:41:27,760 --> 00:41:30,960 YEAR AND A HALF AGO AND ASKED 918 00:41:30,960 --> 00:41:33,840 WHAT WE DO TO PERFORM GENOME 919 00:41:33,840 --> 00:41:36,520 WIDE SCREENS TO SEE IF WE COULD 920 00:41:36,520 --> 00:41:39,120 FIND ANY GENE THAT COULD KILL 921 00:41:39,120 --> 00:41:45,440 POSITIVE CELLS BUT NOT NEGATIVE 922 00:41:45,440 --> 00:41:50,160 CELLS. 923 00:41:50,160 --> 00:41:53,960 THERE'S NOT MANY CELL LINES 924 00:41:53,960 --> 00:41:54,760 AVAILABLE THAT ARE L POSITIVE 925 00:41:54,760 --> 00:41:56,360 AND YOU HAVE A PROBLEM WITH THE 926 00:41:56,360 --> 00:41:57,680 CONTROL AND WHAT YOU'LL COMPARE 927 00:41:57,680 --> 00:41:58,720 IT TO. 928 00:41:58,720 --> 00:42:01,840 USUALLY SO YOU COULD COMPARE IT 929 00:42:01,840 --> 00:42:04,040 TO S CELLS THE SARCOMA CELL LINE 930 00:42:04,040 --> 00:42:06,240 AND THESE ARE TWO COMPLETELY 931 00:42:06,240 --> 00:42:08,040 DIFFERENT TUMORS. 932 00:42:08,040 --> 00:42:10,160 YOU'RE MOST LIKELY GOING TO FIND 933 00:42:10,160 --> 00:42:12,080 CELL TO CELL VARIATION RATHER 934 00:42:12,080 --> 00:42:13,440 THAN ANYTHING TO YOUR PHENOTYPE 935 00:42:13,440 --> 00:42:17,440 OF INTEREST PATH IN THIS CASE. 936 00:42:17,440 --> 00:42:18,920 WE'VE BEEN THINKING WHAT WE CAN 937 00:42:18,920 --> 00:42:22,480 USE AND MOUSE IS NOT A MODEL TO 938 00:42:22,480 --> 00:42:26,960 USE BECAUSE MICE HAS LONG 939 00:42:26,960 --> 00:42:30,040 TELOMERES THAT NEVER INDUCE ALT 940 00:42:30,040 --> 00:42:35,680 AND WHEN I JOINED NCI MY 941 00:42:35,680 --> 00:42:43,400 NEIGHBOR WSTUDIED DOGS AND 942 00:42:43,400 --> 00:42:46,080 SARCOMA IN DOGS AND DOGS ARE A 943 00:42:46,080 --> 00:42:46,880 BEAUTIFUL SYSTEM FOR CANCER 944 00:42:46,880 --> 00:42:49,520 BIOLOGY AND FOR ALT IN OUR CASE. 945 00:42:49,520 --> 00:42:55,320 THE REASON IS THEY DEVELOP ALSO 946 00:42:55,320 --> 00:42:57,600 SARCOMA AND THERE'S A FRACTION 947 00:42:57,600 --> 00:42:58,720 OF L POSITIVE CELLS. 948 00:42:58,720 --> 00:43:01,120 IN ADDITION TO THIS, TELOMERES 949 00:43:01,120 --> 00:43:04,880 IN DOGS ARE SIMILAR TO US. 950 00:43:04,880 --> 00:43:10,600 ROUGHLY HAVE 10 KILO BASES AND 951 00:43:10,600 --> 00:43:14,320 TELOMERASE SHUTS DOWN LIKE IN US 952 00:43:14,320 --> 00:43:15,520 MAYBE THE DOG IS THE BEST SYSTEM 953 00:43:15,520 --> 00:43:17,880 AND YOU DON'T HAVE TO HOUSE THE 954 00:43:17,880 --> 00:43:18,520 DOGS. 955 00:43:18,520 --> 00:43:21,800 PEOPLE CARE ABOUT DOGS SO THEY 956 00:43:21,800 --> 00:43:23,520 HOUSE THEM THEMSELVES. 957 00:43:23,520 --> 00:43:25,920 SO WE PAIRED WITH AIMY AND WHAT 958 00:43:25,920 --> 00:43:27,560 WE DECIDED TO DO IS FIRST OF ALL 959 00:43:27,560 --> 00:43:32,400 TRY TO SEE CAN WE DETECT ALT AND 960 00:43:32,400 --> 00:43:39,280 POSITIVITY IN TUMOR CELLS OR 961 00:43:39,280 --> 00:43:43,920 CANCER CELLS AND GIANNA TOOK 962 00:43:43,920 --> 00:43:45,480 EVERYTHING AMY GAVE HER AND THIS 963 00:43:45,480 --> 00:43:48,440 IS THE GOLDEN STANDARD FOR 964 00:43:48,440 --> 00:43:53,440 DETECTING THE ROLE IN CYCLE PCR 965 00:43:53,440 --> 00:43:56,480 TO SEE TELOMERIC DNA IN FREE 966 00:43:56,480 --> 00:43:58,520 FLOATING CIRCLE FORM AND WE HAVE 967 00:43:58,520 --> 00:44:01,120 A HUMAN SARCOMA CELL LINE WHICH 968 00:44:01,120 --> 00:44:04,480 IS WHAT ALMOST EVERYBODY USES 969 00:44:04,480 --> 00:44:08,760 WHEN THEY WORK WITH ALT AND WE 970 00:44:08,760 --> 00:44:12,720 HAVE ER POSITIVE CELL LINES IN 971 00:44:12,720 --> 00:44:14,160 HUMANS. 972 00:44:14,160 --> 00:44:17,440 SO THE POSITIVE IS A DARK SIGNAL 973 00:44:17,440 --> 00:44:22,240 FOR THE CELLS AND WE FOUND AND 974 00:44:22,240 --> 00:44:25,400 GIANNA FOUND TUMOR SAMPLES THAT 975 00:44:25,400 --> 00:44:28,480 ARE L POSITIVE IN THE DOGS AS 976 00:44:28,480 --> 00:44:31,360 WELL AS CANCER CELL LINES 977 00:44:31,360 --> 00:44:34,200 PRIMARY CANCER CELL LINE DERIVED 978 00:44:34,200 --> 00:44:36,480 FROM DOGS FROM THE TUMORS THAT 979 00:44:36,480 --> 00:44:40,080 ARE L POSITIVE AND A LOT OF L 980 00:44:40,080 --> 00:44:41,480 NEGATIVE SAMPLES. 981 00:44:41,480 --> 00:44:45,400 SO THE BEAUTY OF THIS IS AMY 982 00:44:45,400 --> 00:44:48,240 ALSO COLLECTED A LOT OF 983 00:44:48,240 --> 00:44:49,400 TRANSCRIPTIONAL DATA FROM THE 984 00:44:49,400 --> 00:44:49,840 SAME CELLS. 985 00:44:49,840 --> 00:44:50,760 SO WE HAVE A LOT OF INFORMATION 986 00:44:50,760 --> 00:44:51,720 ABOUT THIS. 987 00:44:51,720 --> 00:44:52,760 WE HAVE INFORMATION IN TUMORS AS 988 00:44:52,760 --> 00:44:54,960 WELL AS IN THE CELL LINES. 989 00:44:54,960 --> 00:44:57,840 SO NOW WE ARE IN THE PERFECT 990 00:44:57,840 --> 00:45:00,240 POSITION TO ASK AND SORRY, THE 991 00:45:00,240 --> 00:45:01,400 ADDITIONAL POSITIVE AND NICE 992 00:45:01,400 --> 00:45:04,120 THING ABOUT DOGS IS GENETICALLY 993 00:45:04,120 --> 00:45:07,360 THEY'RE CLOSE TO EACH OTHER THAN 994 00:45:07,360 --> 00:45:08,320 WE ARE. 995 00:45:08,320 --> 00:45:09,560 OKAY. 996 00:45:09,560 --> 00:45:11,960 SO BASICALLY NOW WE ARE IN THE 997 00:45:11,960 --> 00:45:12,680 PERFECT SITUATION OR SHE'S IN 998 00:45:12,680 --> 00:45:13,840 THE PERFECT SITUATION TO PERFORM 999 00:45:13,840 --> 00:45:14,760 THE SCREEN. 1000 00:45:14,760 --> 00:45:18,520 OF COURSE WE NEEDED A DOG, 1001 00:45:18,520 --> 00:45:21,080 GENOME WIDE LIBRARY AND WE WERE 1002 00:45:21,080 --> 00:45:23,880 IN THE PROCESS OF MAKING THIS 1003 00:45:23,880 --> 00:45:26,320 OURSELVES BUT WE WERE LUCKY TO 1004 00:45:26,320 --> 00:45:28,040 FIND A COLLABORATOR THAT 1005 00:45:28,040 --> 00:45:29,240 GENERATED THE LIBRARY AND GAVE 1006 00:45:29,240 --> 00:45:31,800 IT TO US AND NOW WE HAVE THE 1007 00:45:31,800 --> 00:45:32,200 LIBRARY. 1008 00:45:32,200 --> 00:45:36,160 SO NOW EVERYTHING IS SET TO 1009 00:45:36,160 --> 00:45:38,440 SCREEN THIS CANCER CELL LINES. 1010 00:45:38,440 --> 00:45:41,160 DOG CANCER CELL LINES AND FIND 1011 00:45:41,160 --> 00:45:42,520 ANYTHING THAT IS ESSENTIAL IN 1012 00:45:42,520 --> 00:45:43,960 POSITIVE BUT NOT L NEGATIVE 1013 00:45:43,960 --> 00:45:44,480 CELLS. 1014 00:45:44,480 --> 00:45:45,800 AND OF COURSE THE FINAL GOAL OF 1015 00:45:45,800 --> 00:45:50,040 THE PROJECT WOULD BE THEN TO 1016 00:45:50,040 --> 00:45:51,600 CROSS THIS WITH SCREENS IN HUMAN 1017 00:45:51,600 --> 00:45:53,600 CELLS AND WITH THE GENETIC DATA 1018 00:45:53,600 --> 00:45:58,240 OUT THERE WITH HUMAN CELLS AND 1019 00:45:58,240 --> 00:45:59,280 POTENTIALLY FIND COMMON TIPS. 1020 00:45:59,280 --> 00:46:03,440 SO WITH THIS I CONCLUDE JUST BY 1021 00:46:03,440 --> 00:46:07,720 AGAIN REMINDING YOU WHAT I TOLD 1022 00:46:07,720 --> 00:46:08,000 YOU. 1023 00:46:08,000 --> 00:46:12,440 I TOLD YOU HOW TELOMERES ARE 1024 00:46:12,440 --> 00:46:15,320 PROTECTIVE AND MAYBE IN CANCER 1025 00:46:15,320 --> 00:46:17,440 CELLS TO OVERCOME PROBLEMS WITH 1026 00:46:17,440 --> 00:46:17,720 TELOMERES. 1027 00:46:17,720 --> 00:46:19,360 I DIDN'T TELL YOU HOW IT WORKS 1028 00:46:19,360 --> 00:46:22,480 BUT WHAT WE'RE DOING TO TRY TO 1029 00:46:22,480 --> 00:46:25,400 UNDERSTAND THE MECHANISM OF 1030 00:46:25,400 --> 00:46:28,360 TELOMERE ELONGATION IN CANCER 1031 00:46:28,360 --> 00:46:29,480 CELLS. 1032 00:46:29,480 --> 00:46:30,760 IF YOU'RE INTERESTED IN THIS, 1033 00:46:30,760 --> 00:46:33,440 COME TALK TO ME AND I LOVE 1034 00:46:33,440 --> 00:46:36,400 TALKING ABOUT TELOMERES AND 1035 00:46:36,400 --> 00:46:37,400 ANYTHING ELSE. 1036 00:46:37,400 --> 00:46:39,000 AND I TRIED TO THANK EVERYBODY 1037 00:46:39,000 --> 00:46:40,200 IN THE LAB FOR THE DATA I 1038 00:46:40,200 --> 00:46:40,480 SHOWED. 1039 00:46:40,480 --> 00:46:46,040 I COULDN'T SHOW THE WORK OF SOME 1040 00:46:46,040 --> 00:46:52,040 BUT I WANT TO SHOW I HAVE THIS 1041 00:46:52,040 --> 00:46:53,280 AMAZING GROUP OF PEOPLE THAT 1042 00:46:53,280 --> 00:46:54,040 KEEPS WORKING IN A DIFFICULT 1043 00:46:54,040 --> 00:46:55,560 TIME AND IT'S BEEN A PLEASURE TO 1044 00:46:55,560 --> 00:47:00,480 WORK WITH THEM AND IN ADDITION 1045 00:47:00,480 --> 00:47:02,400 THERE'S A LOT OF PEOPLE THAT I 1046 00:47:02,400 --> 00:47:03,880 SHOULD THANK AND IT'S A 1047 00:47:03,880 --> 00:47:05,520 FANTASTIC PLACE IT DO SCIENCE. 1048 00:47:05,520 --> 00:47:07,400 THANK YOU. 1049 00:47:07,400 --> 00:47:14,440 THANK YOU. 1050 00:47:14,440 --> 00:47:16,480 >>UNFORTUNATELY WE CAN'T TAKE 1051 00:47:16,480 --> 00:47:20,360 QUESTIONS FROM THE PEOPLE THAT 1052 00:47:20,360 --> 00:47:23,800 ARE OUTSIDE BUT HAPPY TO TAKE 1053 00:47:23,800 --> 00:47:24,480 QUESTIONS. 1054 00:47:24,480 --> 00:47:25,560 >>THANK YOU. 1055 00:47:25,560 --> 00:47:28,000 THE DOG MODEL'S REALLY 1056 00:47:28,000 --> 00:47:28,320 ATTRACTIVE. 1057 00:47:28,320 --> 00:47:31,720 IT'S UNIQUE TO NCI SO GOOD 1058 00:47:31,720 --> 00:47:31,960 RESULT. 1059 00:47:31,960 --> 00:47:36,440 SINCE HAVE THE GENOMICS, I'M 1060 00:47:36,440 --> 00:47:38,880 CURIOUS WHETHER THE TUMORS ALT 1061 00:47:38,880 --> 00:47:43,080 BY THE PCR BASE WHETHER THEY 1062 00:47:43,080 --> 00:47:43,760 LOSE THE EXPRESSION. 1063 00:47:43,760 --> 00:47:45,760 I LOOKED AT THE GENES YOU HAD ON 1064 00:47:45,760 --> 00:47:48,200 YOUR LIST BUT THERE WAS NO CHART 1065 00:47:48,200 --> 00:47:51,760 AND I WOULD ASSUME THE TUMOR IS 1066 00:47:51,760 --> 00:47:54,400 TELOMERASE POSITIVE IT SHOULD 1067 00:47:54,400 --> 00:47:55,800 HAVE TERT POSITIVITY IN THE 1068 00:47:55,800 --> 00:47:59,560 EXPRESSION AND THE ALT SHOULD BE 1069 00:47:59,560 --> 00:48:00,520 TERT NEGATIVE HAVE YOU LOOKED AT 1070 00:48:00,520 --> 00:48:01,560 THE TERT IN THE TUMORS. 1071 00:48:01,560 --> 00:48:04,400 >>I'VE BEEN TOLD I HAVE TO 1072 00:48:04,400 --> 00:48:05,040 REPEAT THE QUESTION FOR THE 1073 00:48:05,040 --> 00:48:06,160 PEOPLE ONLINE. 1074 00:48:06,160 --> 00:48:09,840 SO THE QUESTION IS BASICALLY IN 1075 00:48:09,840 --> 00:48:14,560 THE ALT TUMORS -- SORRY, IN THE 1076 00:48:14,560 --> 00:48:16,680 DOG TUMORS IF WE COMPARE 1077 00:48:16,680 --> 00:48:19,080 EXPRESSION LEVEL DID WE FIND 1078 00:48:19,080 --> 00:48:20,240 TELOMERASE AND EXPRESSION IN THE 1079 00:48:20,240 --> 00:48:21,880 POSITIVE AND NEGATIVE. 1080 00:48:21,880 --> 00:48:26,680 IN THE CASE OF THE DOG, I DIDN'T 1081 00:48:26,680 --> 00:48:28,440 MINE THE DATA SO THE DATA I HAVE 1082 00:48:28,440 --> 00:48:30,160 IS WHAT A SHOWED YOU BUT YOU 1083 00:48:30,160 --> 00:48:30,960 SHOULD BE ABLE TO. 1084 00:48:30,960 --> 00:48:36,160 IF YOU GO TO DETMAP AND LOOK AT 1085 00:48:36,160 --> 00:48:40,280 THE -- TRY TO STRATIFY TUMORS 1086 00:48:40,280 --> 00:48:42,120 BASED ON THE TELOMERE LENGTH, 1087 00:48:42,120 --> 00:48:46,640 TELOMERASE SAY -- IS A GOOD WAY 1088 00:48:46,640 --> 00:48:49,440 TO DO IT AND IT'S NOT EXPRESSED 1089 00:48:49,440 --> 00:48:50,760 HIGHLY SO YOU'LL HAVE TUMORS 1090 00:48:50,760 --> 00:48:53,440 THAT CANNOT BE CHARACTERIZED. 1091 00:48:53,440 --> 00:48:56,080 YOU'LL FIND THE BOTTOM 1% AND 1092 00:48:56,080 --> 00:48:57,520 EVERYTHING IN THE MIDDLE WILL BE 1093 00:48:57,520 --> 00:48:57,880 QUESTIONABLE. 1094 00:48:57,880 --> 00:48:59,320 >>THE REASON I'M ASKING IS 1095 00:48:59,320 --> 00:49:00,960 BECAUSE I LOOKED AT THE CANCER 1096 00:49:00,960 --> 00:49:02,920 CELL LINES AND IT'S VERY CLEAR 1097 00:49:02,920 --> 00:49:04,480 THIS WAS BLACK AND WHITE. 1098 00:49:04,480 --> 00:49:07,360 WE CAN DISCUSS FURTHER TOGETHER 1099 00:49:07,360 --> 00:49:09,320 BUT IT WOULD BE NICE TO MINE THE 1100 00:49:09,320 --> 00:49:10,600 GENOMIC DATA FROM THE DOGS AND 1101 00:49:10,600 --> 00:49:12,480 SEE IF THAT CAN BE USED BECAUSE 1102 00:49:12,480 --> 00:49:16,000 IT COULD BE AN EASY WAY TO DO 1103 00:49:16,000 --> 00:49:17,920 RNA SEQ AND WHAT IS POSITIVE AND 1104 00:49:17,920 --> 00:49:19,760 NEGATIVE AND THEN WHAT YOU SAID 1105 00:49:19,760 --> 00:49:21,440 IS IT THERAPEUTIC AND SOMETHING 1106 00:49:21,440 --> 00:49:22,200 WE CAN DEAL WITH. 1107 00:49:22,200 --> 00:49:23,240 >>FOR SURE. 1108 00:49:23,240 --> 00:49:27,680 THAT'S A GOOD WAY. 1109 00:49:27,680 --> 00:49:29,200 IT'S NOT -- AGAIN, THERE'S A 1110 00:49:29,200 --> 00:49:33,080 GRAY AREA AND WHEN I DID DATA 1111 00:49:33,080 --> 00:49:35,320 MINING ON HUMAN CELLS AND I'M 1112 00:49:35,320 --> 00:49:37,000 NOT SURE HOW TO CHARACTERIZE. 1113 00:49:37,000 --> 00:49:37,960 THERE'S A LOT OF TUMORS IN THE 1114 00:49:37,960 --> 00:49:39,240 MIDDLE. 1115 00:49:39,240 --> 00:49:43,720 >>THANKS. 1116 00:49:43,720 --> 00:49:45,200 >>HI, EROS. 1117 00:49:45,200 --> 00:49:48,000 WHAT DO YOU THINK IS UNIQUE TO 1118 00:49:48,000 --> 00:49:49,680 THE EMBRYONIC CELLS THAT ALLOWS 1119 00:49:49,680 --> 00:49:51,600 THEM OR WHAT ADVANTAGE THEY HAVE 1120 00:49:51,600 --> 00:49:53,440 FOR NOT REGULATING THEIR 1121 00:49:53,440 --> 00:49:55,080 TELOMERES IN THE SAME WAY AND 1122 00:49:55,080 --> 00:49:56,920 ALSO DO YOU THINK THE SAME 1123 00:49:56,920 --> 00:49:59,680 MECHANISM IS AT PLAY IN HUMAN 1124 00:49:59,680 --> 00:50:02,040 EMBRYOS OR EMBRYONIC CELLS? 1125 00:50:02,040 --> 00:50:04,240 >>AGAIN, I'M REPEAT. 1126 00:50:04,240 --> 00:50:07,400 THE QUESTION IS ABOUT BASICALLY 1127 00:50:07,400 --> 00:50:10,280 IT'S WHAT IS UNIQUE ABOUT THE 1128 00:50:10,280 --> 00:50:12,160 CELLS AND WHY THEY DON'T SEEM TO 1129 00:50:12,160 --> 00:50:13,640 CARE SO MUCH ABOUT THIS 1130 00:50:13,640 --> 00:50:14,560 PROTECTIVE PROTEINS AND THE 1131 00:50:14,560 --> 00:50:17,760 SECOND PART OF THE QUESTION IS 1132 00:50:17,760 --> 00:50:19,400 WHETHER HUMAN CELLS BEHAVE IN 1133 00:50:19,400 --> 00:50:20,080 THE SAME WAY. 1134 00:50:20,080 --> 00:50:21,800 SO THE SECOND QUESTION IS ARE 1135 00:50:21,800 --> 00:50:23,040 REALLY EASY TO ANSWER AND THE 1136 00:50:23,040 --> 00:50:23,920 ANSWER IS I DON'T KNOW. 1137 00:50:23,920 --> 00:50:28,320 WE DIDN'T DO IT SO WORKING ON 1138 00:50:28,320 --> 00:50:29,680 CELLS HERE IT'S CHALLENGING. 1139 00:50:29,680 --> 00:50:33,640 SO WE STARTED A COLLABORATION 1140 00:50:33,640 --> 00:50:36,440 WITH A LAB IN WASH-U AND SO FAR 1141 00:50:36,440 --> 00:50:40,680 HE HASN'T BEEN ABLE TO GENERATE 1142 00:50:40,680 --> 00:50:42,720 TR2 KNOCKOUT CELLS BUT I CANNOT 1143 00:50:42,720 --> 00:50:47,160 KNOW IF THAT'S BECAUSE YOU CAN'T 1144 00:50:47,160 --> 00:50:48,480 OR -- SO THE FIRST QUESTION IS 1145 00:50:48,480 --> 00:50:50,360 THE MILLION-DOLLAR QUESTION, 1146 00:50:50,360 --> 00:50:52,080 LIKE WHY. 1147 00:50:52,080 --> 00:50:52,840 AND I DON'T KNOW. 1148 00:50:52,840 --> 00:50:57,040 THE TRUTH IS I DON'T KNOW. 1149 00:50:57,040 --> 00:50:59,960 IT COULD BE ONE POTENTIAL ISSUE 1150 00:50:59,960 --> 00:51:03,680 THERE IS THE CELLS, A, REALLY 1151 00:51:03,680 --> 00:51:04,960 SHORT SO THE CELLS DON'T STICK 1152 00:51:04,960 --> 00:51:06,960 AROUND VERY LONG. 1153 00:51:06,960 --> 00:51:09,680 WE ARTIFICIALLY BLOCK THEM IN 1154 00:51:09,680 --> 00:51:11,320 THIS SITUATION AND DURING EMBRYO 1155 00:51:11,320 --> 00:51:15,440 GENESIS WE HAVE THE BOOST OF 1156 00:51:15,440 --> 00:51:17,280 PROLIFERA 1157 00:51:17,280 --> 00:51:17,600 PROLIFERATION. 1158 00:51:17,600 --> 00:51:19,160 YOU MAY WANT TO FIND WAYS TO NOT 1159 00:51:19,160 --> 00:51:22,160 DEAL WITH THE CONVOLUTED WAY OF 1160 00:51:22,160 --> 00:51:22,760 PROTECTING TELOMERES FOR 1161 00:51:22,760 --> 00:51:24,520 INSTANCE IF YOU'RE THINKING OF 1162 00:51:24,520 --> 00:51:28,200 SECONDARY STRUCTURES AND THAT 1163 00:51:28,200 --> 00:51:29,400 COULD BE ONE EXPLANATION. 1164 00:51:29,400 --> 00:51:31,400 THE OTHER COULD BE INSTEAD OF 1165 00:51:31,400 --> 00:51:33,160 BEING A TELOMERE SPECIFIC THING 1166 00:51:33,160 --> 00:51:34,560 WE'RE SEEING, MAYBE WE'RE JUST 1167 00:51:34,560 --> 00:51:35,080 LOOKING AT SOMETHING MORE 1168 00:51:35,080 --> 00:51:38,280 GLOBAL. 1169 00:51:38,280 --> 00:51:40,200 FOR INSTANCE, THE CELLS ARE 1170 00:51:40,200 --> 00:51:43,040 KNOWN TO HAVE A WEIRD, LET'S 1171 00:51:43,040 --> 00:51:44,280 SAY, DNA DAMAGE RESPONSE. 1172 00:51:44,280 --> 00:51:46,680 IT COULD BE IN GENERAL THEY 1173 00:51:46,680 --> 00:51:48,920 DON'T RESPOND AS NICELY TO DNA 1174 00:51:48,920 --> 00:51:50,000 DAMAGED CELLS. 1175 00:51:50,000 --> 00:51:53,440 DOESN'T SEEM TO BE TOTALLY TRUE 1176 00:51:53,440 --> 00:51:55,240 FOR DOUBLE STRAND BREAKS AND 1177 00:51:55,240 --> 00:51:57,200 HAVE CANONICAL DNA RESPONSE BUT 1178 00:51:57,200 --> 00:51:57,880 THERE MAY BE THINGS WE DON'T 1179 00:51:57,880 --> 00:51:59,600 SEE. 1180 00:51:59,600 --> 00:52:04,040 >>BEAUTIFUL STORY. 1181 00:52:04,040 --> 00:52:05,320 THIS MAY BE A NAIVE QUESTION. 1182 00:52:05,320 --> 00:52:10,400 I WAS STRUCK BY THE PHENOTYPIC 1183 00:52:10,400 --> 00:52:12,480 DIFFERENCE BETWEEN THE CELLS WHY 1184 00:52:12,480 --> 00:52:16,840 YOU SEE LONG STRINGS OF INFUSION 1185 00:52:16,840 --> 00:52:20,080 AND YOU HAVE TWO OR THREE 1186 00:52:20,080 --> 00:52:22,360 CHROMOSOMES FUSING DO YOU THINK 1187 00:52:22,360 --> 00:52:23,800 IT'S A GAIN OF FUNCTION 1188 00:52:23,800 --> 00:52:25,000 PHENOTYPE OR LOSS OF FUNCTION 1189 00:52:25,000 --> 00:52:28,320 PHENOTYPE BECAUSE THE CRISPR 1190 00:52:28,320 --> 00:52:30,200 SCREEN SUGGESTS LOSS OF FUNCTION 1191 00:52:30,200 --> 00:52:32,560 GIVEN THE BIOLOGY BUT IS THAT A 1192 00:52:32,560 --> 00:52:34,800 REASON TO THINK THAT OR IS THERE 1193 00:52:34,800 --> 00:52:36,200 A POSSIBILITY THERE'S SOMETHING 1194 00:52:36,200 --> 00:52:38,760 NEW IN THE DIFFERENTIATED CELLS? 1195 00:52:38,760 --> 00:52:41,280 >>SO THE QUESTION HAS TO DO 1196 00:52:41,280 --> 00:52:43,120 WITH PRETTY MUCH THE AMOUNT OF 1197 00:52:43,120 --> 00:52:43,720 FUSIONS WE SEE. 1198 00:52:43,720 --> 00:52:47,320 SO IN THE SOMATIC CELLS WE SEE 1199 00:52:47,320 --> 00:52:49,240 LONG STRAINS OF SPAGHETTI LIKE 1200 00:52:49,240 --> 00:52:52,520 FUSIONS AND IN THE ESL CELLS WE 1201 00:52:52,520 --> 00:52:55,680 CAB GET FUSIONS WE GET FEW. 1202 00:52:55,680 --> 00:53:00,760 SO WHY IS THAT? 1203 00:53:00,760 --> 00:53:04,000 AND DON'T KNOW. 1204 00:53:04,000 --> 00:53:05,800 WE DON'T HAVE THE PROOF YET 1205 00:53:05,800 --> 00:53:07,760 HOWEVER, I THINK THERE'S A 1206 00:53:07,760 --> 00:53:09,600 COUPLE THINGS. 1207 00:53:09,600 --> 00:53:13,440 SO THE DIFFUSION ARE MAINLY 1208 00:53:13,440 --> 00:53:16,760 OCCURRING IN G1. 1209 00:53:16,760 --> 00:53:21,000 SO IT COULD BE IN THE EMBRYONIC 1210 00:53:21,000 --> 00:53:22,480 STEM CELLS WHEN YOU GET THE 1211 00:53:22,480 --> 00:53:25,040 FUSION FROM THE CRISPR SCREEN 1212 00:53:25,040 --> 00:53:27,400 THEY'RE STILL OCCURRING TUMORS 1213 00:53:27,400 --> 00:53:31,480 BUT THE G1 AND S CELLS ARE 1214 00:53:31,480 --> 00:53:32,920 RELATIVELY SHORT TO SOMATIC 1215 00:53:32,920 --> 00:53:33,120 CELLS. 1216 00:53:33,120 --> 00:53:34,840 YOU MAY NOT HAVE IF YOU HAVE 1217 00:53:34,840 --> 00:53:35,120 TIME. 1218 00:53:35,120 --> 00:53:37,720 IF YOU THINK ABOUT IT FROM A 1219 00:53:37,720 --> 00:53:38,360 PROBABILITY POINT OF VIEW YOU 1220 00:53:38,360 --> 00:53:40,520 HAVE TO FIND A PROTECTED 1221 00:53:40,520 --> 00:53:42,400 TELOMERE YOU HAVE TO FIND 1222 00:53:42,400 --> 00:53:43,960 ANOTHER AND AND ANOTHER ONE AND 1223 00:53:43,960 --> 00:53:45,080 ANOTHER ONE YOU MAY NEED MORE 1224 00:53:45,080 --> 00:53:47,520 TIME FOR THIS TO HAPPEN. 1225 00:53:47,520 --> 00:53:50,720 IT MAY BE A BORING EXPLANATION 1226 00:53:50,720 --> 00:53:52,440 BUT HOWEVER, IT COULD BE THERE'S 1227 00:53:52,440 --> 00:53:53,880 SOMETHING ELSE. 1228 00:53:53,880 --> 00:53:55,560 IDEALLY THAT'S WHY WE KEEP DOING 1229 00:53:55,560 --> 00:53:56,960 THE SCREENS. 1230 00:53:56,960 --> 00:54:01,000 IDEALLY I'D LIKE TO GET TO THE 1231 00:54:01,000 --> 00:54:03,200 POINT WHERE EVERYTHING FUSES AND 1232 00:54:03,200 --> 00:54:05,760 THEN I'D BE SATISFIED I FOUND 1233 00:54:05,760 --> 00:54:06,040 EVERYTHING. 1234 00:54:06,040 --> 00:54:08,400 >>THANK YOU. 1235 00:54:08,400 --> 00:54:10,520 >>I HAVE A QUICK QUESTION. 1236 00:54:10,520 --> 00:54:15,280 FIRST QUESTION OUT OF CURIOSITY 1237 00:54:15,280 --> 00:54:19,440 HAVE YOU LOOKED FOR THINGS THAT 1238 00:54:19,440 --> 00:54:23,080 ARE ESSENTIAL IN WILD TYPE CELLS 1239 00:54:23,080 --> 00:54:26,440 BUT STOP BEING ESSENTIAL IN 1240 00:54:26,440 --> 00:54:28,080 HUMAN CELLS PROTECTED. 1241 00:54:28,080 --> 00:54:29,400 >>THE QUESTION IS WHERE ARE WE 1242 00:54:29,400 --> 00:54:32,640 IN ADDITION TO LOOKING FOR 1243 00:54:32,640 --> 00:54:43,160 SYNTHETIC AND LOOKING FOR GENES 1244 00:54:46,000 --> 00:54:49,240 SYNTHETICALLY BOUND WITH THE 1245 00:54:49,240 --> 00:54:50,400 CELL TYPES. 1246 00:54:50,400 --> 00:54:53,440 AND IF YOU NOTICE BASICALLY THE 1247 00:54:53,440 --> 00:54:55,520 ESL CELLS WHEN WE DELETE EVEN IF 1248 00:54:55,520 --> 00:54:58,720 THEY SURVIVE AND DON'T GROW AS 1249 00:54:58,720 --> 00:54:59,840 WELL AS THE WILD TYPE. 1250 00:54:59,840 --> 00:55:01,680 ONE OF THE REASON WE DID THE 1251 00:55:01,680 --> 00:55:03,080 SECOND SCREEN WAS ALSO FINDING 1252 00:55:03,080 --> 00:55:03,760 THINGS THAT WOULD MAKE THE CELLS 1253 00:55:03,760 --> 00:55:06,360 GROW FASTER. 1254 00:55:06,360 --> 00:55:07,960 THAT COULD GIVE AN INDICATE OF 1255 00:55:07,960 --> 00:55:10,120 WHAT WAS HAPPENING. 1256 00:55:10,120 --> 00:55:13,560 AND WE DID FIND SOME GENES SO A 1257 00:55:13,560 --> 00:55:19,400 LOT OF CHROMATIN MODIFYING GENES 1258 00:55:19,400 --> 00:55:21,400 BUT DIDN'T FOLLOW-UP AND WE 1259 00:55:21,400 --> 00:55:23,840 DIDN'T VERIFY IT. 1260 00:55:23,840 --> 00:55:27,200 WE DON'T KNOW IF THAT'S THE CASE 1261 00:55:27,200 --> 00:55:28,280 BUT IT'S INTERESTING. 1262 00:55:28,280 --> 00:55:30,440 >>IT'S VERY INTERESTING. 1263 00:55:30,440 --> 00:55:32,360 AND SECOND QUICK QUESTION SO TO 1264 00:55:32,360 --> 00:55:33,600 FIND THE PROTEINS THAT HAVE 1265 00:55:33,600 --> 00:55:36,720 TELOMERE TO FIND THE ENDS OF 1266 00:55:36,720 --> 00:55:38,040 CHROMOSOMES YOU DID LIGATION 1267 00:55:38,040 --> 00:55:42,960 WITH TPP1 BUT WHY COULDN'T YOU 1268 00:55:42,960 --> 00:55:45,160 JUST USE TELOMERE BASED PROTEIN. 1269 00:55:45,160 --> 00:55:48,120 >>THE QUESTION IS FOR THE 1270 00:55:48,120 --> 00:55:51,000 PROTEOMICS WHY GO WITH TPP1 1271 00:55:51,000 --> 00:55:52,760 RATHER THAN TELOMERASE. 1272 00:55:52,760 --> 00:55:55,400 THE ANSWER IS TARGETING 1273 00:55:55,400 --> 00:55:58,000 TELOMERASE IS TRICKY. 1274 00:55:58,000 --> 00:55:59,560 YOU CAN AFFECT DYSFUNCTION. 1275 00:55:59,560 --> 00:56:01,760 THERE'S A LOT OF REPORTS ON THAT 1276 00:56:01,760 --> 00:56:05,640 SO WE DID NOT WANT TO TOUCH 1277 00:56:05,640 --> 00:56:07,080 TELOMERASE IF POSSIBLE. 1278 00:56:07,080 --> 00:56:08,080 >>OKAY. 1279 00:56:08,080 --> 00:56:10,640 >>SO THE OTHER THING WE 1280 00:56:10,640 --> 00:56:13,120 CONSIDERED DOING WAS INSTEAD 1281 00:56:13,120 --> 00:56:13,800 TARGETING THE RNA. 1282 00:56:13,800 --> 00:56:18,280 YOU CAN TAG THE RNA COMPONENT, 1283 00:56:18,280 --> 00:56:20,480 YOU KNOW AND THE FACTORS THERE 1284 00:56:20,480 --> 00:56:22,080 BUT TPP1 WAS THE EASIEST WAY TO 1285 00:56:22,080 --> 00:56:24,000 DO IT. 1286 00:56:24,000 --> 00:56:24,280 >>I SEE. 1287 00:56:24,280 --> 00:56:29,640 THANK YOU. 1288 00:56:29,640 --> 00:56:32,960 >>JUST WITH YOUR VERY COOL 1289 00:56:32,960 --> 00:56:36,480 ASSAY FOR MUTANT TELOMERASE 1290 00:56:36,480 --> 00:56:41,080 ADDITION, CAN YOU -- HAVE PEOPLE 1291 00:56:41,080 --> 00:56:43,920 STUDIED THE PROCESS THE CRISIS 1292 00:56:43,920 --> 00:56:47,720 THAT THEN LEADS TO RE-EXPRESSION 1293 00:56:47,720 --> 00:56:50,680 OF TELOMERASE AND BECAUSE THAT'S 1294 00:56:50,680 --> 00:56:52,560 WHAT HAPPENS RIGHT AFTER THE 1295 00:56:52,560 --> 00:56:54,360 PROCESS YOU GET RE-EXPRESSION 1296 00:56:54,360 --> 00:56:59,880 AND WONDERING ABOUT THE 1297 00:56:59,880 --> 00:57:00,440 FREQUENCY OF THAT IN VITRO. 1298 00:57:00,440 --> 00:57:02,800 >>I SEE -- SO IT'S A GOOD -- SO 1299 00:57:02,800 --> 00:57:04,800 BASICALLY WE -- WE DIDN'T DO IT. 1300 00:57:04,800 --> 00:57:05,800 YOU COULD DO IT. 1301 00:57:05,800 --> 00:57:10,280 SO ONE OF THE ISSUE WITH THAT IS 1302 00:57:10,280 --> 00:57:11,800 IN SOMATIC CELLS EXPRESSION OF 1303 00:57:11,800 --> 00:57:13,840 THAT SO WHENEVER THE MUTANT 1304 00:57:13,840 --> 00:57:14,760 WOULD BE EXPRESSED IT COULD 1305 00:57:14,760 --> 00:57:16,360 BECOME TOXIC. 1306 00:57:16,360 --> 00:57:19,440 SO YOU MIGHT BE SELECTING OUT 1307 00:57:19,440 --> 00:57:19,800 YOUR CELLS. 1308 00:57:19,800 --> 00:57:22,120 BUT IF YOU ONLY LOOK AT 1309 00:57:22,120 --> 00:57:23,600 SHORT-TERM IT COULD BE -- 1310 00:57:23,600 --> 00:57:26,840 >>AND IN YOUR ESLs, DID YOU 1311 00:57:26,840 --> 00:57:30,600 LOOK FOR DEPLETION OF TRF 1312 00:57:30,600 --> 00:57:33,760 PROTEINS OR WOULD THAT TAKE TOO 1313 00:57:33,760 --> 00:57:34,520 LONG BECAUSE THAT'S YOUR 1314 00:57:34,520 --> 00:57:35,560 PREDICTION, RIGHT, YOU WOULDN'T 1315 00:57:35,560 --> 00:57:37,840 HAVE THE BINDING, YOU MENTIONED 1316 00:57:37,840 --> 00:57:39,760 THAT BUT PROBABLY SEE ROBUST 1317 00:57:39,760 --> 00:57:41,040 BINDING. 1318 00:57:41,040 --> 00:57:43,120 >>WE DIDN'T DO LIKE TO SEE IF 1319 00:57:43,120 --> 00:57:47,360 WE COULD FIND THE MUTANTS BUT 1320 00:57:47,360 --> 00:57:50,840 AGAIN WE DIDN'T INVENT THE 1321 00:57:50,840 --> 00:57:53,680 MUTANT RNA ASSAY. 1322 00:57:53,680 --> 00:57:55,960 SO THERE'S A LOT OF WORK DONE IN 1323 00:57:55,960 --> 00:57:57,960 VITRO AND SUPPOSEDLY NONE OF THE 1324 00:57:57,960 --> 00:57:58,600 PROTEINS BIND. 1325 00:57:58,600 --> 00:58:00,880 >>DOES IT DECREASE THE BINDING? 1326 00:58:00,880 --> 00:58:02,360 >>IT SHOULD DECREASE THE 1327 00:58:02,360 --> 00:58:02,560 BINDING. 1328 00:58:02,560 --> 00:58:06,760 WE DID IT IN THE MUTANT. 1329 00:58:06,760 --> 00:58:08,080 >>EVEN IN THE SHORT-TERM 1330 00:58:08,080 --> 00:58:09,560 BECAUSE TELOMERES ARE VERY LONG. 1331 00:58:09,560 --> 00:58:11,880 >>YOU WOULD DO A STAINING YOU 1332 00:58:11,880 --> 00:58:16,960 WOULD STAY AND YOU MAY HAVE 45KB 1333 00:58:16,960 --> 00:58:18,680 OF WILD TYPE AND A LITTLE BIT OF 1334 00:58:18,680 --> 00:58:19,480 DEN. 1335 00:58:19,480 --> 00:58:26,920 >>SO AS YOU SAID THE ALTERNATE 1336 00:58:26,920 --> 00:58:28,280 TELOMERE ASSAY HAS BEEN DONE BY 1337 00:58:28,280 --> 00:58:29,480 OTHERS IN OTHER TYPES OF CELLS. 1338 00:58:29,480 --> 00:58:34,320 I'M WONDERING IF YOU SEE ANY 1339 00:58:34,320 --> 00:58:38,000 CORRELATION BETWEEN THE -- IF 1340 00:58:38,000 --> 00:58:38,960 THERE'S ANY CORRELATION BETWEEN 1341 00:58:38,960 --> 00:58:42,640 THE LENGTH OF THE PRE-EXISTING 1342 00:58:42,640 --> 00:58:47,080 TELOMERES AND THOSE THAT ARE 1343 00:58:47,080 --> 00:58:49,640 CHOSEN TO HAVE NEW SEQUENCES 1344 00:58:49,640 --> 00:58:50,040 ADDED. 1345 00:58:50,040 --> 00:58:52,680 OF COURSE NOT EVERY SPOT WAS 1346 00:58:52,680 --> 00:58:53,880 LABELLED. 1347 00:58:53,880 --> 00:58:57,920 AND WHETHER THAT'S THE SAME IN 1348 00:58:57,920 --> 00:58:58,800 ESL CELLS AS IT IS IN OTHER CELL 1349 00:58:58,800 --> 00:59:00,240 LINES. 1350 00:59:00,240 --> 00:59:01,200 >>YES. 1351 00:59:01,200 --> 00:59:04,560 SO THE QUESTION HAS TO DO WITH 1352 00:59:04,560 --> 00:59:05,960 BASICALLY I'M GOING TO SLIGHTLY 1353 00:59:05,960 --> 00:59:07,040 REPHRASE IT. 1354 00:59:07,040 --> 00:59:11,200 CAN WE USE THE MUTANT TELOMERE 1355 00:59:11,200 --> 00:59:14,320 EASY TO STUDY THE ELONGATION AND 1356 00:59:14,320 --> 00:59:16,760 ACT ON THE SHORTEST TELOMERE 1357 00:59:16,760 --> 00:59:16,920 NOT. 1358 00:59:16,920 --> 00:59:18,840 SO THERE ARE INDICATIONS THAT'S 1359 00:59:18,840 --> 00:59:19,440 THE CASE. 1360 00:59:19,440 --> 00:59:20,200 IN OUR ASSAY WE DIDN'T DO THAT 1361 00:59:20,200 --> 00:59:22,160 YET. 1362 00:59:22,160 --> 00:59:24,400 SO AGAIN, THIS IS FAIRLY NEW. 1363 00:59:24,400 --> 00:59:27,120 BUT YOU CAN DEFINITELY DO IT AND 1364 00:59:27,120 --> 00:59:30,320 ONE THING -- ONE COOL THING WE 1365 00:59:30,320 --> 00:59:34,320 DID WAS TO USE ANDRE'S ASSAY TO 1366 00:59:34,320 --> 00:59:36,600 SEE WHETHER THE MUTANTS ARE AT 1367 00:59:36,600 --> 00:59:40,880 THE VERY END OR NOT OR HOW LONG 1368 00:59:40,880 --> 00:59:42,160 IT IS. 1369 00:59:42,160 --> 00:59:43,440 THINK OF FOR INSTANCE, MEASURING 1370 00:59:43,440 --> 00:59:45,800 THE LENGTH OF THE PROGRESSIVE 1371 00:59:45,800 --> 00:59:46,080 TELOMERASE. 1372 00:59:46,080 --> 00:59:47,160 THESE ARE THINGS WE'RE THINKING 1373 00:59:47,160 --> 00:59:47,560 OF DOING. 1374 00:59:47,560 --> 00:59:48,600 WE'RE IN THE PROCESS OF DOING IT 1375 00:59:48,600 --> 00:59:51,880 BUT THOSE ARE GOOD POINTS. 1376 00:59:51,880 --> 00:59:52,720 YEP. 1377 00:59:52,720 --> 00:59:56,800 >>SO I WANT TO THANK EROS AGAIN 1378 00:59:56,800 --> 00:59:57,760 FOR A BEAUTIFUL TALK AND THE 1379 00:59:57,760 --> 00:59:59,400 KIND OF TALK WHERE YOU WANT TO 1380 00:59:59,400 --> 01:00:00,120 INVITE HIM BACK. 1381 01:00:00,120 --> 01:00:01,400 IT'S PRETTY EXCITING. 1382 01:00:01,400 --> 01:00:02,680 YOU WANT TO KNOW WHAT THE 1383 01:00:02,680 --> 01:00:04,000 RESULTS ARE GOING TO BE IN A 1384 01:00:04,000 --> 01:00:04,760 YEAR, RIGHT. 1385 01:00:04,760 --> 01:00:06,560 I DON'T KNOW WHO ORGANIZES THIS 1386 01:00:06,560 --> 01:00:09,120 BUT CAN WE INVITE HIM BACK? 1387 01:00:09,120 --> 01:00:11,960 GREAT. 1388 01:00:11,960 --> 01:00:12,280 >>THANK YOU. 1389 01:00:12,280 --> 00:00:00,000 >>THANK YOU, THANK YOU.