1 00:00:06,566 --> 00:00:07,467 >> LET'S GET STARTED. 2 00:00:07,467 --> 00:00:09,102 GOT MORNING, GOOD AFTERNOON, 3 00:00:09,102 --> 00:00:10,870 GOOD EVENING EVERYONE EMPLOY 4 00:00:10,870 --> 00:00:16,776 WELCOME TO THE NIH DNA REPAIR 5 00:00:16,776 --> 00:00:18,578 INTEREST GROUP VIDEO 6 00:00:18,578 --> 00:00:18,878 CONFERENCES. 7 00:00:18,878 --> 00:00:22,949 TODAY WE'RE REALLY GLAD TO HAVE 8 00:00:22,949 --> 00:00:24,917 DR. GUO-MIN LI WHO IS A 9 00:00:24,917 --> 00:00:25,752 DISTINGUISHED INVESTIGATOR AND 10 00:00:25,752 --> 00:00:31,057 DIRECTOR FOR THE INSTITUTE OF 11 00:00:31,057 --> 00:00:32,892 CANCER RESEARCH FOR CHINESE 12 00:00:32,892 --> 00:00:36,629 INSTITUTES FOR MEDICAL RESEARCH. 13 00:00:36,629 --> 00:00:38,164 DR. LI RECEIVED HIS Ph.D. FROM 14 00:00:38,164 --> 00:00:42,235 THE UNIVERSITY AND WORKING WITH 15 00:00:42,235 --> 00:00:43,903 [INDISCERNIBLE] IN A POST DOC 16 00:00:43,903 --> 00:00:46,639 FELLOWSHIP, LATER HE STARTED HIS 17 00:00:46,639 --> 00:00:48,441 CAREER IN KENTUCKY UNIVERSITY 18 00:00:48,441 --> 00:00:51,210 AND THEN MOVED TO UNIVERSITY OF 19 00:00:51,210 --> 00:00:54,947 SOUTH CALIFORNIA AND THEN UT 20 00:00:54,947 --> 00:00:55,982 SOUTHWEST MEDICAL CENTER. 21 00:00:55,982 --> 00:00:57,717 HE SERVED AS DISTINGUISHED CHAIR 22 00:00:57,717 --> 00:01:01,921 FOR CANCER RESEARCH AND DIRECTOR 23 00:01:01,921 --> 00:01:03,956 OF TRANSLATIONAL RESEARCH IN 24 00:01:03,956 --> 00:01:09,729 2023 DR. LI MOVED BACK TO CHINA 25 00:01:09,729 --> 00:01:11,831 AND CONTINUE CANCER RESEARCH 26 00:01:11,831 --> 00:01:12,732 INSTITUTE STUDIES. 27 00:01:12,732 --> 00:01:15,268 DR. LI STUDIES THE MECHANISM OF 28 00:01:15,268 --> 00:01:18,037 DNA MISTACH REPAIR AND ITS ROLE 29 00:01:18,037 --> 00:01:20,306 IN CANCER AND HAS MADE SIMILAR 30 00:01:20,306 --> 00:01:21,440 CONTRIBUTION TOWARDS THE 31 00:01:21,440 --> 00:01:22,842 UNDERSTANDING OF THE MECHANISM 32 00:01:22,842 --> 00:01:25,144 BY WHICH THE MISMATCH REPAIR 33 00:01:25,144 --> 00:01:26,078 SYSTEM MAINTAINS GENOME 34 00:01:26,078 --> 00:01:28,514 STABILITY AND CANCER AVOIDANCE. 35 00:01:28,514 --> 00:01:32,251 SO, WITHOUT FURTHER ADO, WE WILL 36 00:01:32,251 --> 00:01:33,653 HAVE DR. LI GIVE THE 37 00:01:33,653 --> 00:01:34,754 PRESENTATION EMPLOY FOR THE 38 00:01:34,754 --> 00:01:37,590 AUDIENCES IF YOU HAVE ANY 39 00:01:37,590 --> 00:01:38,624 QUESTION, PLEASE LEAVE IT IN THE 40 00:01:38,624 --> 00:01:40,860 CHAT BOX AND WE WILL GO THROUGH 41 00:01:40,860 --> 00:01:42,461 AFTER THE SEMINAR. 42 00:01:42,461 --> 00:01:45,431 THANK YOU. 43 00:01:45,431 --> 00:01:47,667 DR. LI? 44 00:01:47,667 --> 00:01:50,870 >> THANK YOU SO MUCH CHUN FOR 45 00:01:50,870 --> 00:01:52,738 THE CANDID INTRODUCTION AND I 46 00:01:52,738 --> 00:01:56,576 WOULD ALSO LIKE TO THANK KAREN 47 00:01:56,576 --> 00:01:57,543 AND [INDISCERNIBLE] FOR PATIENT. 48 00:01:57,543 --> 00:02:00,313 I HAVE BEEN WORKING ON DNA 49 00:02:00,313 --> 00:02:03,916 MISMATCH REPAIR FOR THE PAST 50 00:02:03,916 --> 00:02:04,350 30-SOMETHING YEARS. 51 00:02:04,350 --> 00:02:06,819 I STARTED WITH [INDISCERNIBLE] 52 00:02:06,819 --> 00:02:11,424 OUT OF DUKE UNIVERSITY AT 53 00:02:11,424 --> 00:02:14,393 [INDISCERNIBLE] SO I WILL BE 54 00:02:14,393 --> 00:02:19,365 TODAY, MAINLY FOCUS ON 1 OF THE 55 00:02:19,365 --> 00:02:21,767 KEY MISMATCH REPAIR FACTOR MLH1 56 00:02:21,767 --> 00:02:23,202 NEXT, SLIDE, PLEASE. 57 00:02:23,202 --> 00:02:28,341 SO AS YOU ALL KNOW, THE DNA 58 00:02:28,341 --> 00:02:31,043 MISMATCH REPAIR IS A WAYS DNA 59 00:02:31,043 --> 00:02:32,945 REPLICATION TO INSURE 60 00:02:32,945 --> 00:02:35,481 REPLICATION FREE DAMAGING AND 61 00:02:35,481 --> 00:02:38,184 ALTHOUGH WE KNOW, REPLICATION 62 00:02:38,184 --> 00:02:41,153 PROCESS IS VERY UPDPRAID, EVEN 63 00:02:41,153 --> 00:02:42,455 FROM THE LOCATION AND 64 00:02:42,455 --> 00:02:44,757 OCCASIONALLY MAKE A MISTAKE BY 65 00:02:44,757 --> 00:02:49,262 DOING MISSING CORPORATION WHICH 66 00:02:49,262 --> 00:02:49,896 GENERATE BASE-BASE MISMATCH, 67 00:02:49,896 --> 00:02:55,134 THAT SHOW HERE IN THE MISMATCH 68 00:02:55,134 --> 00:02:56,936 OR INTRODUCE INSERTION DELETION 69 00:02:56,936 --> 00:03:00,873 MISPAIR WITHIN THE SIMPLE 70 00:03:00,873 --> 00:03:04,744 REPETITIVE DNA SEQUENCE, SO THIS 71 00:03:04,744 --> 00:03:10,182 MISPAIRS HAS TO BE CORRECTED TO 72 00:03:10,182 --> 00:03:11,117 AVOID MUTATIONS. 73 00:03:11,117 --> 00:03:12,618 AND UNLIKE OTHER DNA DAMAGES 74 00:03:12,618 --> 00:03:18,591 WHICH ARE VERY OBVIOUS, BUT 75 00:03:18,591 --> 00:03:22,461 MISMATCH REPAIR I MEAN, ACTUALLY 76 00:03:22,461 --> 00:03:24,897 THEIR NORMAL, MOST SPACES ARE 77 00:03:24,897 --> 00:03:28,534 NORMAL DNA COMPOSITION, SO IN 78 00:03:28,534 --> 00:03:30,269 ORDER TO REMOVE [INDISCERNIBLE] 79 00:03:30,269 --> 00:03:32,438 THE MISMATCH REPAIR SYSTEM HAS 80 00:03:32,438 --> 00:03:42,982 TO KNOW WHICH 1 IS 1 AND THIS IS 81 00:03:47,787 --> 00:03:49,555 ACTUALLY PROVIDED BY A SIGNAL IN 82 00:03:49,555 --> 00:03:51,424 THE LIGAND BREAK AND THIS 83 00:03:51,424 --> 00:03:53,793 CONTAINS A FRAG 84 00:03:53,793 --> 00:03:54,493 M-TEBURKEULOSEISOT MEDIAN STRAND 85 00:03:54,493 --> 00:04:00,833 YOU ALSO HAVE THE 3 PAIR FOR THE 86 00:04:00,833 --> 00:04:03,169 END THOSE ENDS OR BREAKS PROVIDE 87 00:04:03,169 --> 00:04:04,670 OPPORTUNITIES FOR MISMATCH 88 00:04:04,670 --> 00:04:08,441 REPAIR TO TARGET THE REPAIR ON 89 00:04:08,441 --> 00:04:09,141 THE NEW [INDISCERNIBLE]. 90 00:04:09,141 --> 00:04:13,112 SO THAT THE MISPAIRED ARE BASED 91 00:04:13,112 --> 00:04:17,817 OR THE INSERTION ON THE 92 00:04:17,817 --> 00:04:20,186 [INDISCERNIBLE] DRIVE OR BOTH 93 00:04:20,186 --> 00:04:22,321 MEDIUM STRAND WILL BE REMOVED TO 94 00:04:22,321 --> 00:04:26,926 INSURE REPLICATION FOR DAMAGING. 95 00:04:26,926 --> 00:04:32,098 NEXT SLIDE. 96 00:04:32,098 --> 00:04:37,370 SO MISMATCH IN HUMAN CELL HAVE 97 00:04:37,370 --> 00:04:39,305 BEEN RECONSITUTED INVITRO USING 98 00:04:39,305 --> 00:04:43,209 ALL PURIFIED PROTEINS BY SEVERAL 99 00:04:43,209 --> 00:04:44,043 LABORATORIES INCLUDING MYSELF 100 00:04:44,043 --> 00:04:52,918 AND [INDISCERNIBLE] AND WE KNOW 101 00:04:52,918 --> 00:04:53,819 THAT [INDISCERNIBLE] PROTEIN IS 102 00:04:53,819 --> 00:04:55,021 THE 1 THAT RECOGNIZE THE 103 00:04:55,021 --> 00:04:58,257 MISMATCH AND THEN LATER ON IT 104 00:04:58,257 --> 00:05:04,930 RECRUITS DUE TO L-PROTEIN TO THE 105 00:05:04,930 --> 00:05:07,333 DNA PROTEIN COMPLEX, AND I'M 106 00:05:07,333 --> 00:05:08,701 SURE [INDISCERNIBLE] LABORATORY 107 00:05:08,701 --> 00:05:11,871 MUTE ANT L-ALPHA IN HUMAN CELLS 108 00:05:11,871 --> 00:05:15,107 HAS THE ENDONUCLEUS ACTIVITY, 109 00:05:15,107 --> 00:05:17,510 AND IT CAN [INDISCERNIBLE] DNA 110 00:05:17,510 --> 00:05:20,379 ON A NEW STRAIN, MODIFIED 111 00:05:20,379 --> 00:05:23,582 PLACES, HOWEVER, WE RECENTLY 112 00:05:23,582 --> 00:05:26,285 SHOWED THAT ONLY IN THE STRAND 113 00:05:26,285 --> 00:05:30,156 BREAK THAT CLOSE THE 2 MISMATCH, 114 00:05:30,156 --> 00:05:31,023 [INDISCERNIBLE] FOR THE MISMATCH 115 00:05:31,023 --> 00:05:34,894 WILL BE SERVED AS A PLACE FOR 116 00:05:34,894 --> 00:05:39,632 AXON 1 RECRUITMENT AND THEN THE 117 00:05:39,632 --> 00:05:42,234 AXON 1 OR STARTING AT THIS 118 00:05:42,234 --> 00:05:44,837 STRAND BREAK AND THEN FROM 119 00:05:44,837 --> 00:05:46,272 [INDISCERNIBLE] TO 3 120 00:05:46,272 --> 00:05:48,007 [INDISCERNIBLE] TO REMOVE THE 121 00:05:48,007 --> 00:05:51,177 MISMATCH, AND THEN ONCE YOU 122 00:05:51,177 --> 00:05:57,483 REACH TO THE MISMATCH, WHERE THE 123 00:05:57,483 --> 00:06:01,787 MUTE ANT HAS A PLACE AND THE 124 00:06:01,787 --> 00:06:04,023 MUTANT SA SLIDING AWAY FROM THE 125 00:06:04,023 --> 00:06:09,195 MISMATCH, SO HERE ARE THE WAY TO 126 00:06:09,195 --> 00:06:11,630 ASK THE NUCLEUS 1 TO REMOVE THE 127 00:06:11,630 --> 00:06:14,400 MISPAIRED BASE, SO THEN THEY 128 00:06:14,400 --> 00:06:17,570 REPLICATE THE POLYMERASE SUCH AS 129 00:06:17,570 --> 00:06:19,505 [INDISCERNIBLE] AND CONJUNCTION 130 00:06:19,505 --> 00:06:23,476 WITH OUR RP A CANNED WHAT AND 131 00:06:23,476 --> 00:06:26,245 THE NEED TO FIND THE 132 00:06:26,245 --> 00:06:29,048 [INDISCERNIBLE] LIAISON GAS 1. 133 00:06:29,048 --> 00:06:33,752 SO THIS COMPLOATS THE REPAIR OF 134 00:06:33,752 --> 00:06:37,423 MISMATCH. 135 00:06:37,423 --> 00:06:38,691 NEXT SLIDE, PLEASE. 136 00:06:38,691 --> 00:06:42,561 SO TODAY, I WILL TRY TO 137 00:06:42,561 --> 00:06:46,432 EMPHASIZE ON HOW MUTANT L-ALPHA 138 00:06:46,432 --> 00:06:49,235 OR 1 OF THE K-MISMATCH REPAIR 139 00:06:49,235 --> 00:06:52,805 COMPONENT WHEN IT CONSISTS OF 140 00:06:52,805 --> 00:06:56,475 MH1, AND PMS2, AND HOW THIS MH01 141 00:06:56,475 --> 00:06:58,377 PLAY A CRITICAL ROLE IN MISMATCH 142 00:06:58,377 --> 00:07:07,019 REPAIR AS WELL AS IN NONMISMATCH 143 00:07:07,019 --> 00:07:14,793 REPAIR FUNCTION, NEXT SLIDE IT 144 00:07:14,793 --> 00:07:22,968 WAS THE FIRST MISMATCH REPAIR 145 00:07:22,968 --> 00:07:25,604 IDENTIFIED WHICH CAN ACTUALLY, 146 00:07:25,604 --> 00:07:31,744 AND THE IDENTIFICATION OF MUTANT 147 00:07:31,744 --> 00:07:34,346 L-ALPHA AND THE MUTANT BEATA AND 148 00:07:34,346 --> 00:07:37,483 THE OTHER HOMOLOGUES. 149 00:07:37,483 --> 00:07:37,950 NEXT SLIDE. 150 00:07:37,950 --> 00:07:40,619 SO THIS IS THE WAYS 151 00:07:40,619 --> 00:07:41,453 IDENTIFICATION OF 152 00:07:41,453 --> 00:07:43,689 [INDISCERNIBLE] IS THE STABILITY 153 00:07:43,689 --> 00:07:46,659 IN COLORECTAL CANCER BY 154 00:07:46,659 --> 00:07:49,795 DR. [INDISCERNIBLE] LABORATORY 155 00:07:49,795 --> 00:07:52,631 AS WELL AS 3 OTHER LABORATORY 156 00:07:52,631 --> 00:07:54,266 INCLUDING [INDISCERNIBLE] AT 157 00:07:54,266 --> 00:07:54,800 MAYO CLINIC. 158 00:07:54,800 --> 00:08:01,740 SO WHAT THEY SHOWED IN THEIR 159 00:08:01,740 --> 00:08:06,278 STUDY IS THAT THEY FIND TUMOR 160 00:08:06,278 --> 00:08:09,481 CELLS DISPLAY SIMPLE REPETITIVE 161 00:08:09,481 --> 00:08:11,450 SEQUENCE IN COMPARISON WITH 162 00:08:11,450 --> 00:08:16,088 NORMAL CELLS AND AS YOU CAN SEE 163 00:08:16,088 --> 00:08:18,924 IN THIS PICTURE THAT TUMOR CELL 164 00:08:18,924 --> 00:08:21,293 EITHER THEY HAVE EXPANSION OF 165 00:08:21,293 --> 00:08:22,127 SIMPLE REPETITIVE DNA SEQUENCE 166 00:08:22,127 --> 00:08:26,298 OR THEY HAVE A CONFRACTION, 167 00:08:26,298 --> 00:08:28,834 CONTRACTING THE SIMPLE 168 00:08:28,834 --> 00:08:29,401 REPETITIVE DNA SEQUENCE. 169 00:08:29,401 --> 00:08:32,738 SO AT THAT TIME, THEY ONLY 170 00:08:32,738 --> 00:08:35,674 KNOW--THEY DON'T KNOW EXACTLY 171 00:08:35,674 --> 00:08:37,076 WHAT'S HAPPENING, THEY CONCLUDE 172 00:08:37,076 --> 00:08:40,913 THIS IS DUE TO REPLICATION ARROW 173 00:08:40,913 --> 00:08:46,352 SO YOU CALL IT THE RER+ OR THE 174 00:08:46,352 --> 00:08:47,353 RER- TUMOR PLEASE. 175 00:08:47,353 --> 00:08:50,389 SO WE HAVE WAYS THIS, I MEAN 176 00:08:50,389 --> 00:08:51,757 PUBLICATIONS, AND THE WAY WE 177 00:08:51,757 --> 00:08:55,361 BELIEVE THAT PROBABLY RELATED TO 178 00:08:55,361 --> 00:08:58,163 MISMATCH REPAIR DEFACT, SO WE 179 00:08:58,163 --> 00:09:02,134 GOT A CELL ON THE 180 00:09:02,134 --> 00:09:05,771 [INDISCERNIBLE] MINOR CELLS FROM 181 00:09:05,771 --> 00:09:09,942 THE VOGELSTEIN AS WELL AS HIS 182 00:09:09,942 --> 00:09:17,650 POST DOC RAMON PARSONS AND WE 183 00:09:17,650 --> 00:09:18,951 GOT THE HRER+ H6, AND WE DIDN'T 184 00:09:18,951 --> 00:09:20,386 KNOW WHAT IT WAS AND AT THE TIME 185 00:09:20,386 --> 00:09:25,658 WE WERE TOLD THAT IT WAS A CELL 186 00:09:25,658 --> 00:09:26,292 GENERATED FROM [INDISCERNIBLE] 187 00:09:26,292 --> 00:09:28,761 LABORATORY AND LATER ON WE KNOW 188 00:09:28,761 --> 00:09:29,928 THIS ACTUALLY THE 189 00:09:29,928 --> 00:09:31,864 [INDISCERNIBLE] 6, SO IT WAS 190 00:09:31,864 --> 00:09:34,767 SHORT FOR H6, SO WE MEASURED A 191 00:09:34,767 --> 00:09:37,703 MISMEASURED REPAIR ACTIVITY IN 192 00:09:37,703 --> 00:09:44,777 BASE H6 AND SO WHICH STANDS FOR 193 00:09:44,777 --> 00:09:47,579 S WO480, AS CAN YOU SEE IN THIS 194 00:09:47,579 --> 00:09:50,749 MISMATCH REPAIR ASSAY, THE H6 195 00:09:50,749 --> 00:09:56,055 CELL LINE CANNOT GENERATE TOO 196 00:09:56,055 --> 00:09:57,890 SMALL FRAGMENTS, THE REPAIR PULL 197 00:09:57,890 --> 00:10:00,893 BACK OR SO CAN DO VERY WELL, AND 198 00:10:00,893 --> 00:10:03,395 AT LEAST 50% REPAIR ACTIVITY, 199 00:10:03,395 --> 00:10:08,600 YOU CAN SEE IN SO, SO THIS SHOWS 200 00:10:08,600 --> 00:10:11,837 FOR THE FIRST TIME THAT HUMAN 201 00:10:11,837 --> 00:10:17,609 CELL TUMOR CELL DISPLAY, 202 00:10:17,609 --> 00:10:19,545 REPLICATION DEFECTIVE IN THE 203 00:10:19,545 --> 00:10:23,415 MISMATCH REPAIR. 204 00:10:23,415 --> 00:10:24,116 NEXT SLIDES PLEASE. 205 00:10:24,116 --> 00:10:27,519 SO WE THEN KNOW THAT THIS MUST 206 00:10:27,519 --> 00:10:30,155 BE MISSING SOMETHING, RIGHT? 207 00:10:30,155 --> 00:10:31,023 CRITICAL MISMATCH REPAIR 208 00:10:31,023 --> 00:10:31,323 COMPONENT. 209 00:10:31,323 --> 00:10:36,028 SO I STARTED TO TRY TO IDENTIFY 210 00:10:36,028 --> 00:10:38,997 OR PIEWNIFY ACTIVITY TO RESTORE 211 00:10:38,997 --> 00:10:40,833 MISMATCH REPAIR TO H6 CELL LINES 212 00:10:40,833 --> 00:10:46,038 AND AS YOU CAN SEE, IN THE LEFT, 213 00:10:46,038 --> 00:10:53,011 THIS IS PUTIFICATION PROFILE. 214 00:10:53,011 --> 00:11:00,419 THERE WAS A VERY SHARP PEEK WITH 215 00:11:00,419 --> 00:11:02,388 DO THE EXTRACT AND THIS WE PUT 216 00:11:02,388 --> 00:11:04,256 ALL THE DATA TOGETHER AND THEN 217 00:11:04,256 --> 00:11:07,893 PUBLISHED THIS WORK IN CELL, AND 218 00:11:07,893 --> 00:11:10,229 THAT WAS A LONG TIME AGO, SO 30 219 00:11:10,229 --> 00:11:20,672 YEARS AGO AND NEXT SLIDE, 220 00:11:28,847 --> 00:11:29,248 PLEASE. 221 00:11:29,248 --> 00:11:33,986 SO I TARTED WORKING ON THE 222 00:11:33,986 --> 00:11:38,023 PURIFY THIS AND I PURE PIE THIS 223 00:11:38,023 --> 00:11:38,657 ACTIVITY THROUGH CHROMATOGRAPHYS 224 00:11:38,657 --> 00:11:43,829 AND AS YOU CAN SEE IN THE RIGHT 225 00:11:43,829 --> 00:11:46,498 GEL AND THOSE ARE 2 BANDS, I 226 00:11:46,498 --> 00:11:52,805 REALLY CANNOT SEPARATE THIS FOR 227 00:11:52,805 --> 00:11:53,038 THAT. 228 00:11:53,038 --> 00:11:55,140 AND THEN WE HAD NO IDEA WHAT 229 00:11:55,140 --> 00:11:57,075 THIS COMPONENT IS, BECAUSE 230 00:11:57,075 --> 00:11:58,944 COMPARED WITH THE 231 00:11:58,944 --> 00:12:01,814 [INDISCERNIBLE] OR MISMATCH 232 00:12:01,814 --> 00:12:04,183 REPAIR COMPONENT THE E.COLI ARE 233 00:12:04,183 --> 00:12:07,820 FULL OF SINGLE PEPTIDE, SO 1 234 00:12:07,820 --> 00:12:09,221 [INDISCERNIBLE] DILL O DALTON 235 00:12:09,221 --> 00:12:15,461 PROTEIN AND THE MUTANT IS ALSO 236 00:12:15,461 --> 00:12:18,497 SINGLE POLYPEPTIDE, SO IN ORDER 237 00:12:18,497 --> 00:12:22,201 TO KNOW WHAT THIS 2 BANDS ARE, 238 00:12:22,201 --> 00:12:26,171 SO WE PERFORMED A SEQUENCE AND 239 00:12:26,171 --> 00:12:27,339 ASSAYS ON DEGRADATION AND THAT 240 00:12:27,339 --> 00:12:32,277 YOU CAN SEE IN THIS TABLE AND WE 241 00:12:32,277 --> 00:12:34,746 FOUND AND WE FOUND THE PEPTIDES 242 00:12:34,746 --> 00:12:36,181 FROM THESE 2. 243 00:12:36,181 --> 00:12:39,051 THEY ARE HEREWITH HOMOLOGY WAYS 244 00:12:39,051 --> 00:12:42,254 OR HUMAN AT THAT TIME, 245 00:12:42,254 --> 00:12:46,091 IDENTIFIED AND ALSO HAS SOME 246 00:12:46,091 --> 00:12:50,829 YEAST YEENS AND THEN THE BIGGER 247 00:12:50,829 --> 00:12:53,131 1, ALTHOUGH THEY WERE--WE FOUND 248 00:12:53,131 --> 00:12:56,201 THERE WERE SEVERAL, SEVERAL OF 249 00:12:56,201 --> 00:12:57,970 THIS GENES BY AT LEAST WE KNOW 250 00:12:57,970 --> 00:13:01,073 THIS PROBABLY JUST COME FROM A 251 00:13:01,073 --> 00:13:09,715 SINGLE, MAYBE A SINGLE 252 00:13:09,715 --> 00:13:11,683 POLYPEPTIDE IN PMS2, SO BOTH OF 253 00:13:11,683 --> 00:13:19,791 THIS PMS2 AND THE MLH1 HAD THE 254 00:13:19,791 --> 00:13:21,193 SAME PHENOTYPE, SHOWING THE SAME 255 00:13:21,193 --> 00:13:23,595 PHENOTYPE, WE BELIEVE THIS 256 00:13:23,595 --> 00:13:25,364 PROBABLY MUTANT L HYDRODIMER SO 257 00:13:25,364 --> 00:13:27,499 WE PUBLISHED THIS WORK AS SHOWN 258 00:13:27,499 --> 00:13:30,769 THIS IS FOR A HUMAN MUTANT, AND 259 00:13:30,769 --> 00:13:40,212 IT IS A HYDRODIMER AND THE NEXT 260 00:13:40,212 --> 00:13:42,748 SLIDES PLEASE EMPLOY SO AS I 261 00:13:42,748 --> 00:13:44,616 TOLD YOU ABOUT THIS MUTANT EXTRA 262 00:13:44,616 --> 00:13:46,919 HELP TO DEFINE THE HUMAN 263 00:13:46,919 --> 00:13:49,121 MISMATCH REPAIR POSSIBLY, THAT'S 264 00:13:49,121 --> 00:13:51,657 AS WELL AS IDENTIFICATION OF 265 00:13:51,657 --> 00:13:53,191 HUMAN MUTANTS, THE REASON I SAY 266 00:13:53,191 --> 00:14:00,599 THAT, NEXT SLIDE, PLEASE EMPLOY 267 00:14:00,599 --> 00:14:04,903 I ACTUALLY PURE PIED A 268 00:14:04,903 --> 00:14:07,573 HYDRODIMER 6 MONTHS AT 269 00:14:07,573 --> 00:14:09,441 DR. [INDISCERNIBLE] LABORATORY 270 00:14:09,441 --> 00:14:10,342 AND THIS HYDRODIMERRIZED SHOWN 271 00:14:10,342 --> 00:14:19,318 ON THE RIGHT, THIS 160 AND 105 272 00:14:19,318 --> 00:14:21,286 KILODALTON RATINGS AND WE, I 273 00:14:21,286 --> 00:14:30,028 PURIFIED THIS USING AFFINITY 274 00:14:30,028 --> 00:14:31,530 COLUMN, OLIGO CAN CONTAIN A 275 00:14:31,530 --> 00:14:32,931 SINGLE MISMATCH AND I COUPLE 276 00:14:32,931 --> 00:14:34,766 THEM TOGETHER AND THEY ARE 277 00:14:34,766 --> 00:14:38,737 COUPLED TOGETHER, WITH THE DNA 278 00:14:38,737 --> 00:14:39,571 MOLECULES INTO [INDISCERNIBLE] 279 00:14:39,571 --> 00:14:41,306 AND WE ARE PAST THE 280 00:14:41,306 --> 00:14:44,343 [INDISCERNIBLE] YOU COULD 281 00:14:44,343 --> 00:14:46,511 EXTRACT THE SMOOTHEST COLUMN, 282 00:14:46,511 --> 00:14:49,715 AND IT DID 1 PART, IT DOESN'T 283 00:14:49,715 --> 00:14:57,456 MATTER, I MEAN HOW--IT'S THE 284 00:14:57,456 --> 00:14:59,324 WHOLE CELL EXTRACT, THE 285 00:14:59,324 --> 00:15:01,026 [INDISCERNIBLE] JUST PURIFIED 286 00:15:01,026 --> 00:15:02,427 THE FRACTIONS AND THE CANNED 287 00:15:02,427 --> 00:15:04,930 WHAT THEY CAN GET IN THESE 2 288 00:15:04,930 --> 00:15:09,301 BANS AND I WAS SO EXCITED BY THE 289 00:15:09,301 --> 00:15:11,169 DATA I COULD SHARE THAT 1 IS NO 290 00:15:11,169 --> 00:15:13,872 WAY--I MEAN IF YOU THINK OF THIS 291 00:15:13,872 --> 00:15:20,078 HAS A MISMATCH, WE GOTTA SAY, 1 292 00:15:20,078 --> 00:15:21,680 BAND, [INDISCERNIBLE] SO I'M 293 00:15:21,680 --> 00:15:23,415 STUDYING WORKING ON THIS FOR 294 00:15:23,415 --> 00:15:25,951 ANOTHER 6 MONTH, I REALLY CANNOT 295 00:15:25,951 --> 00:15:28,620 SEVER THIS 2 BAND, SO AT THIS 296 00:15:28,620 --> 00:15:31,923 TIME, WE CANNOT REALLY DEFINE 297 00:15:31,923 --> 00:15:32,691 WHAT IT IS. 298 00:15:32,691 --> 00:15:36,228 SO WE JUST PUT A PUTATIVE 299 00:15:36,228 --> 00:15:37,629 PROTEIN IN THE FREEZER AND 300 00:15:37,629 --> 00:15:39,831 WORKING ON SOMETHING ELSE. 301 00:15:39,831 --> 00:15:44,302 AND FROM THIS STUDY, I REALLY, I 302 00:15:44,302 --> 00:15:46,638 MEAN, LEARNED HOW TO DO SCIENCE 303 00:15:46,638 --> 00:15:47,272 FROM DR. [INDISCERNIBLE]. 304 00:15:47,272 --> 00:15:52,644 SO IF YOU ARE NOT A 305 00:15:52,644 --> 00:15:53,578 [INDISCERNIBLE] ON SOMETHING, 306 00:15:53,578 --> 00:15:54,980 DON'T PUBLISH, TRY TO FIGURE IT 307 00:15:54,980 --> 00:15:56,615 OUT BEFORE YOU PUBLISH THIS 308 00:15:56,615 --> 00:15:57,416 STUFF. 309 00:15:57,416 --> 00:16:00,052 SO BECAUSE OF THE IDENTIFICATION 310 00:16:00,052 --> 00:16:03,955 OF MUTANT L ALPHA WHICH IS THE 311 00:16:03,955 --> 00:16:06,258 HETERODIMER AND THEN THIS 312 00:16:06,258 --> 00:16:07,926 IMMEDIATELY MUST KNOW THAT THE 313 00:16:07,926 --> 00:16:09,528 MAYBE THE 1 I PURIFIED 4 YEARS 314 00:16:09,528 --> 00:16:13,565 AGO, IT IS NOT THE HYDRODIMER, 315 00:16:13,565 --> 00:16:18,537 AND THAT INDEED, WE FOUND FROM 316 00:16:18,537 --> 00:16:20,806 BOTH THE [INDISCERNIBLE] 317 00:16:20,806 --> 00:16:23,975 ANTIBODIES ALREADY, AVAILABLE, 318 00:16:23,975 --> 00:16:26,945 SO WE TO INVEST IN 319 00:16:26,945 --> 00:16:31,750 [INDISCERNIBLE] AND SHOWING THAT 320 00:16:31,750 --> 00:16:32,584 [INDISCERNIBLE] BAND 105 WAS 321 00:16:32,584 --> 00:16:35,954 [INDISCERNIBLE] AND THEN BY 322 00:16:35,954 --> 00:16:39,424 SEQUENCING ANALYSIS, WE FOUND 323 00:16:39,424 --> 00:16:40,592 THAT [INDISCERNIBLE] AND AT THIS 324 00:16:40,592 --> 00:16:44,463 TIME WE DON'T KNOW WHAT IS MHS6, 325 00:16:44,463 --> 00:16:47,032 THAT WE NAME OUR 160 KILODALTON 326 00:16:47,032 --> 00:16:49,568 PROTEIN WHICH IS A MUTANT S 327 00:16:49,568 --> 00:16:51,436 ALPHA, WE NAMED IN THIS SCIENCE 328 00:16:51,436 --> 00:16:58,577 PAPER. 329 00:16:58,577 --> 00:16:59,010 NEXT SLIDE, PLEASE. 330 00:16:59,010 --> 00:17:00,612 AND IN ADDITION TO THAT, I 331 00:17:00,612 --> 00:17:03,048 SHOWED HERE, AND ALL THE MUTANT 332 00:17:03,048 --> 00:17:04,483 S CELLS ARE HYDRODIMERS, THIS 333 00:17:04,483 --> 00:17:06,985 ALL STARTED A WAYS 334 00:17:06,985 --> 00:17:09,488 IDENTIFICATION OF MUTANT L 335 00:17:09,488 --> 00:17:13,258 ALPHA, WHETHER I SHOWED YOU 336 00:17:13,258 --> 00:17:15,527 ALREADY, THE [INDISCERNIBLE] FOR 337 00:17:15,527 --> 00:17:18,597 THE DIMER AND THERE ARE ALMOST, 338 00:17:18,597 --> 00:17:24,469 ALL ASSOCIATED WITH THE CANCER 339 00:17:24,469 --> 00:17:26,338 OR OTHER DISEASES. 340 00:17:26,338 --> 00:17:28,206 THAT'S NOT--I'M NOT GOING TO 341 00:17:28,206 --> 00:17:31,076 TALK ABOUT MSH3, WHICH IS REALLY 342 00:17:31,076 --> 00:17:35,247 INVOLVED IN NEURON DEGENERATIVE 343 00:17:35,247 --> 00:17:35,781 DISEASES. 344 00:17:35,781 --> 00:17:39,317 NEXT SLIDE, PLEASE. 345 00:17:39,317 --> 00:17:43,188 SO WHILE I'M GOING TO FOCUS ON 346 00:17:43,188 --> 00:17:46,758 MUTANT L-ALPHA ACTIVITY AGAIN, 347 00:17:46,758 --> 00:17:51,763 MUTANT L-ALPHA CONTAINS A LOT OF 348 00:17:51,763 --> 00:17:55,333 SIMILAR BIOCHEMICAL ACTIVITIES 349 00:17:55,333 --> 00:17:57,302 INCLUDING ATP ASE ACTIVITY WHICH 350 00:17:57,302 --> 00:18:02,808 WAS INITIALLY SHOWED BY DR. YANG 351 00:18:02,808 --> 00:18:07,145 IN E.COLI AND LATER 352 00:18:07,145 --> 00:18:08,313 DR. [INDISCERNIBLE] LABORATORY 353 00:18:08,313 --> 00:18:10,816 SHOWING HUMAN L-PROTEIN AS ATP 354 00:18:10,816 --> 00:18:16,621 ASE ACTIVITY AND I MENTIONED 355 00:18:16,621 --> 00:18:17,556 EARLIER DR. [INDISCERNIBLE] 356 00:18:17,556 --> 00:18:19,524 ENDONUCLEUS ACTIVITY OF MUSEUM 357 00:18:19,524 --> 00:18:21,493 TABT L-ALPHA WHICH IS CRITICAL 358 00:18:21,493 --> 00:18:28,099 TO PROVIDE THE INITIAL STRAND 359 00:18:28,099 --> 00:18:30,101 BREAK TO RECRUIT ALE 1 TO THE 360 00:18:30,101 --> 00:18:32,938 SIDE TO REMOVE THE MISMATCH. 361 00:18:32,938 --> 00:18:36,007 AND THEN TODAY I'M GOING TO SHOW 362 00:18:36,007 --> 00:18:40,245 YOU 1 OPPORTUNITY WE IDENTIFIED 363 00:18:40,245 --> 00:18:45,450 AFTER THIS MUTANT L-ALPHTHIS 364 00:18:45,450 --> 00:18:47,886 ACTIVITY LIKELY REGULATING THE 365 00:18:47,886 --> 00:18:48,987 AXON NUCLEUS 1, ACTIVITY AND I 366 00:18:48,987 --> 00:18:53,992 WILL SHOW YOU HOW THIS NUCLEUS 367 00:18:53,992 --> 00:18:57,529 ACTIVITY INVOLVED IN THE 368 00:18:57,529 --> 00:19:00,832 MUTANT-LOR MISMATCH REPAIR 369 00:19:00,832 --> 00:19:04,002 DEFICIENCY, MEDIATED CANCER 370 00:19:04,002 --> 00:19:06,905 THERAPY AND ALSO THE--INVOLVED 371 00:19:06,905 --> 00:19:09,641 IN THE NEURON DEGENERATIVE 372 00:19:09,641 --> 00:19:15,981 DISEASE INCLUDING HUNTING TON 373 00:19:15,981 --> 00:19:18,149 DISEASE. 374 00:19:18,149 --> 00:19:18,917 NEXT SLIDE. 375 00:19:18,917 --> 00:19:20,485 SO THIS ACTUALLY SHOW YOU, I 376 00:19:20,485 --> 00:19:24,990 TRIED TO SHOW YOU HOW I 377 00:19:24,990 --> 00:19:33,899 IDENTIFIED THIS MUTEITANT - 378 00:19:33,899 --> 00:19:34,332 -MUTANT L-ALPHA 379 00:19:34,332 --> 00:19:36,401 ACTIVITY AND DURING THE 380 00:19:36,401 --> 00:19:37,736 CONSTITUTION OF MISMATCH REPAIR 381 00:19:37,736 --> 00:19:40,038 REACTION AND I SHOWED IT HERE WE 382 00:19:40,038 --> 00:19:42,107 USE ALL PURIFIED PROTEIN AND 383 00:19:42,107 --> 00:19:45,343 THAT THE RIGHT 1 SHOWS THE 384 00:19:45,343 --> 00:19:48,880 MISMATCH REPAIR ASSAY USING ALL 385 00:19:48,880 --> 00:19:52,417 PURIFIED PROTEIN EMPLOY AS CAN 386 00:19:52,417 --> 00:19:55,854 YOU SEE IN 9-4 WHEN WE TAKING 387 00:19:55,854 --> 00:19:58,857 OUTTA A MUTANT L-ALPHA IN THIS 388 00:19:58,857 --> 00:20:03,228 REACTION, AND YOU CAN SEE, THAT 389 00:20:03,228 --> 00:20:05,530 DEALING ACTUALLY, THE TOTAL 390 00:20:05,530 --> 00:20:07,532 AMOUNT IS NARNLGINAL IN THIS 391 00:20:07,532 --> 00:20:09,601 REACTION COMPARE WITH OTHERS, 392 00:20:09,601 --> 00:20:10,635 EITHER THE UNREPAIRED POP BAND 393 00:20:10,635 --> 00:20:17,175 OR IN THE EVEN THE REPAIR, THE 394 00:20:17,175 --> 00:20:19,210 [INDISCERNIBLE] DIED USING THE 395 00:20:19,210 --> 00:20:20,612 SMALLEST FRAGMENT SO WE WANT TO 396 00:20:20,612 --> 00:20:22,247 FIND OUT WHAT IS HAPPENING IN 397 00:20:22,247 --> 00:20:24,683 THIS REACTION. 398 00:20:24,683 --> 00:20:25,583 NEXT, SLIDE, PLEASE. 399 00:20:25,583 --> 00:20:28,820 SO WE MEASURED IN THE LEFT, CAN 400 00:20:28,820 --> 00:20:32,691 YOU SEE, ACTUALLY, IN THE 401 00:20:32,691 --> 00:20:34,125 [INDISCERNIBLE], YOU CAN SEE 402 00:20:34,125 --> 00:20:40,265 AGAIN, THE DNA MOLECULES, AMOUNT 403 00:20:40,265 --> 00:20:41,566 OF MOLECULES, MUCH, MUCH WORSE 404 00:20:41,566 --> 00:20:44,302 COMPARED WITH OTHER ACTIONS 405 00:20:44,302 --> 00:20:48,940 BECAUSE THIS REACTION IS NO 406 00:20:48,940 --> 00:20:50,108 MUTANT L-ALPHA AND WE FINDALLY 407 00:20:50,108 --> 00:20:52,177 FIND OUT THIS IS PROBABLY DUE TO 408 00:20:52,177 --> 00:20:54,579 THE REASON THAT THE EXCISION 409 00:20:54,579 --> 00:20:59,551 STUDY FROM THE STRAND BREAK AND 410 00:20:59,551 --> 00:21:04,956 THE AXLE NUCLEUS WORKED FROM TOP 411 00:21:04,956 --> 00:21:08,293 TO THE BOTTOM AND THEN FOR THIS 412 00:21:08,293 --> 00:21:10,128 IS [INDISCERNIBLE] ASSAYS, YOU 413 00:21:10,128 --> 00:21:15,033 CAN SEE, ACTUALLY REMOVED ON THE 414 00:21:15,033 --> 00:21:17,302 SIDE FOR THE PROBE AND THAT'S 415 00:21:17,302 --> 00:21:25,076 WHY WE COULD BT DETECT THE 416 00:21:25,076 --> 00:21:26,711 MORNING CREWS ANYMORE, SO IT 417 00:21:26,711 --> 00:21:28,279 SHOWS THE AMOUNT OF DNA SO IN 418 00:21:28,279 --> 00:21:29,781 THE RIGHT, WHAT I WANT TO SHOW 419 00:21:29,781 --> 00:21:33,284 YOU IS THAT IN THE PRESENCE OF 420 00:21:33,284 --> 00:21:38,823 MUTANT L-ALPHA IN THE LEFT, 421 00:21:38,823 --> 00:21:40,992 THE--YOU CAN SEE [INDISCERNIBLE] 422 00:21:40,992 --> 00:21:45,063 IS IMMEDIATELY TERMINATED WHEN 423 00:21:45,063 --> 00:21:49,467 THE EXO1 REMOVES THE MISMATCH. 424 00:21:49,467 --> 00:21:51,903 AND ON THE RIGHT WHEN YOU DON'T 425 00:21:51,903 --> 00:21:56,841 HAVE A MUTANT L-PROTEIN, OR IN 426 00:21:56,841 --> 00:21:58,510 ABSENCE OF MLH1, THE EXCISION IS 427 00:21:58,510 --> 00:22:02,047 VERY EXTENSIVE AND THIS MUTANT 428 00:22:02,047 --> 00:22:04,382 L, THIS MUTANT L TO STOP THE 429 00:22:04,382 --> 00:22:07,385 NUCLEUS ACTIVITY, SO WE GENERATE 430 00:22:07,385 --> 00:22:09,654 LARGE AMOUNT OF SINGLE STRANDED 431 00:22:09,654 --> 00:22:11,022 DNA, WHICH COULD BE A PROBLEM AS 432 00:22:11,022 --> 00:22:21,499 I'M GOING TO SHOW YOU LATER, 433 00:22:24,102 --> 00:22:24,936 NEXT SLIDE, PLEASE. 434 00:22:24,936 --> 00:22:27,906 JUST ACTIVITY IN THE WORLD IN 435 00:22:27,906 --> 00:22:28,573 CANCER IMMUNOTHERAPY. 436 00:22:28,573 --> 00:22:37,415 AND THAT WE KNOW IN 1999-2015, 437 00:22:37,415 --> 00:22:40,485 IT HAS BEEN SHARED BY SEVERAL 438 00:22:40,485 --> 00:22:45,924 LABORATORY THAT MISMATCH REPAIR 439 00:22:45,924 --> 00:22:46,391 DEFICIENCY BENEFITS 440 00:22:46,391 --> 00:22:50,795 IMMUNOTHERAPY AND THIS HAS BEEN 441 00:22:50,795 --> 00:22:58,136 REPUTED IN ANIMAL MODEL AS WELL. 442 00:22:58,136 --> 00:22:58,803 NEXT SLIDE. 443 00:22:58,803 --> 00:23:03,741 AND THEN THE MECHANISM AT THAT 444 00:23:03,741 --> 00:23:05,110 TIME WAS--ESSENTIALLY CONDITION 445 00:23:05,110 --> 00:23:07,412 CLUEDED THIS SLIDE, SO IF YOU 446 00:23:07,412 --> 00:23:09,514 DON'T HAVE A MISMATCH REPAIR 447 00:23:09,514 --> 00:23:13,685 ACTIVITY, YOU ARE GOING TO BE 448 00:23:13,685 --> 00:23:16,154 MUTATE A LOT OF MUTATIONS, THE 449 00:23:16,154 --> 00:23:19,757 FRAMING SHIFT OF MUTATIONS WHICH 450 00:23:19,757 --> 00:23:21,593 CAN CAUSE EITHER SHORTER 451 00:23:21,593 --> 00:23:23,094 PEPTIDE, BECAUSE YOU MAY 452 00:23:23,094 --> 00:23:26,664 GENERATE A STOP CODON OR YOU 453 00:23:26,664 --> 00:23:32,737 CODING FOR SOMETHING THAT IS NOT 454 00:23:32,737 --> 00:23:36,674 ALSO NEEDED, THERE IS NOT A NEED 455 00:23:36,674 --> 00:23:39,210 THIS PROTEIN, SO THIS PROTEIN 456 00:23:39,210 --> 00:23:42,080 WILL BE DEGRADED WHICH WILL 457 00:23:42,080 --> 00:23:46,484 GENERATE MORE PEPTIDE, SO THAT'S 458 00:23:46,484 --> 00:23:49,354 GOING TO USE ITS NEOANTIGEN, SO 459 00:23:49,354 --> 00:23:51,890 THE IDEA IS THAT THE CANCER 460 00:23:51,890 --> 00:23:55,193 CELL, OR MISMATCH REPAIR 461 00:23:55,193 --> 00:23:57,962 DEFICIENT TUMOR CELL WILL NOT 462 00:23:57,962 --> 00:23:59,597 HAVE A NEOANTIGEN WHICH WILL NOT 463 00:23:59,597 --> 00:24:04,602 BE RECOGNIZED BY OUR T-CELL SO 464 00:24:04,602 --> 00:24:07,105 THAT THIS MUTATED TUMOR CELL 465 00:24:07,105 --> 00:24:09,174 WILL BE TREATED AS A FOREIGN, A 466 00:24:09,174 --> 00:24:13,845 FOREIGN AND THEN PROBABLY GOING 467 00:24:13,845 --> 00:24:16,581 TO BE KILLED BY T-CELL. 468 00:24:16,581 --> 00:24:19,884 NEXT SLIDE. 469 00:24:19,884 --> 00:24:20,718 INTERESTINGLY ALTHOUGH THIS 470 00:24:20,718 --> 00:24:22,086 EXPERIMENTATION IS REALLY NICE, 471 00:24:22,086 --> 00:24:26,157 I MEAN IT CAN EXPLAIN AT LEAST 472 00:24:26,157 --> 00:24:30,895 50% OF CASES THAT PATIENTS 473 00:24:30,895 --> 00:24:32,163 RESPONDED TO IMMUNOTHERAPY, BUT 474 00:24:32,163 --> 00:24:36,000 OTHER 50% OF TUMORS, MISMATCH 475 00:24:36,000 --> 00:24:39,671 REPAIR DEFICIENT TUMORS, THEY DO 476 00:24:39,671 --> 00:24:41,539 HAVE NEOANTIGEN, THEY DO NOT 477 00:24:41,539 --> 00:24:44,609 HAVE RESPONDED TO PD1, PDL 478 00:24:44,609 --> 00:24:49,647 ANTIBODY IMMUNOTHERAPY AND 479 00:24:49,647 --> 00:24:51,716 SUGGESTING SOMETHING ELSE MUST 480 00:24:51,716 --> 00:24:54,252 ENROLL IN THIS, SO WE ARE REALLY 481 00:24:54,252 --> 00:24:56,154 INTERESTED IN THIS AND WE TRY TO 482 00:24:56,154 --> 00:25:02,694 FIND THE ISO FOR THIS. 483 00:25:02,694 --> 00:25:03,595 NEXT SLIDE. 484 00:25:03,595 --> 00:25:05,530 SO THEN GOING INTO THE 485 00:25:05,530 --> 00:25:08,399 LITERATURE AND FIND THAT GENOME 486 00:25:08,399 --> 00:25:12,170 INSTABILITY IS ASSOCIATE WIDE 487 00:25:12,170 --> 00:25:15,306 THE DNA PRODUCTION AND THE CGAS 488 00:25:15,306 --> 00:25:18,943 EVALUATION, SO THIS IS A REVIEW 489 00:25:18,943 --> 00:25:21,012 BY MY FORMER COLLEAGUE JIM CHEN 490 00:25:21,012 --> 00:25:22,247 AND HEITATED HERE THAT IF YOU 491 00:25:22,247 --> 00:25:24,182 HAVE A DNA BREAKS OR CHROMOSOME 492 00:25:24,182 --> 00:25:28,453 BREAKS, YOU ARE ANYTHING TO 493 00:25:28,453 --> 00:25:29,487 GENERATE [INDISCERNIBLE] DNA AND 494 00:25:29,487 --> 00:25:31,823 IT WILL BE RECOGNIZED BY CGAS, 495 00:25:31,823 --> 00:25:42,333 CGAS WILL BE ABLE TO GENERATE 496 00:25:48,239 --> 00:25:50,642 CCAMP--HELPING IMMUNOTHERAPY AS 497 00:25:50,642 --> 00:25:54,579 HONE BY CHEN IN ANIMAL MODEL, IF 498 00:25:54,579 --> 00:26:00,184 HE SHOWS THAT CGAS IS INVOLVED 499 00:26:00,184 --> 00:26:03,154 IN PD1, PDL1, PLOK TREATMENT, I 500 00:26:03,154 --> 00:26:08,993 MEAN TRY TO KILL CANCER, CANCER 501 00:26:08,993 --> 00:26:09,193 CELLS. 502 00:26:09,193 --> 00:26:11,095 SO WE BELIEVE THAT A MISMATCH 503 00:26:11,095 --> 00:26:14,165 REPAIR DEFICIENCY CAN INDUCE 504 00:26:14,165 --> 00:26:17,835 GENOMIC INSTABILITY THAT TRIGGER 505 00:26:17,835 --> 00:26:20,672 CYTOSOLIC DNA [INDISCERNIBLE] 506 00:26:20,672 --> 00:26:22,807 AND ACTIVATE CGAS POSSIBLY 507 00:26:22,807 --> 00:26:25,743 IMMUNOTHERAPY, THIS IS WORK BY 508 00:26:25,743 --> 00:26:27,945 [INDISCERNIBLE] AND THIS WORK WE 509 00:26:27,945 --> 00:26:38,456 HAVE PUBLISHED A COUPLE YEARS 510 00:26:59,811 --> 00:27:03,448 AGO, NEXT SLIDE. 511 00:27:03,448 --> 00:27:09,587 CELLS SHOWING THE DNA BUT ONCE 512 00:27:09,587 --> 00:27:11,923 YOU PUT MHL1 BACK AND REDUCED A 513 00:27:11,923 --> 00:27:16,327 LOT, DOESN'T MATTER, YOU 514 00:27:16,327 --> 00:27:18,629 JUSTIFIED CELL ITSELF OR TREAT 515 00:27:18,629 --> 00:27:21,399 IT WITH RADIATION WHICH CAN CAN 516 00:27:21,399 --> 00:27:24,369 TRIGGER A LOT OF DNA PRODUCTION 517 00:27:24,369 --> 00:27:27,772 BUT WHEN YOU PUT IT THE MH1 518 00:27:27,772 --> 00:27:30,041 BACK, THE AMOUNT IS DRAMATICALLY 519 00:27:30,041 --> 00:27:35,813 ARE I DUCED NEXT SLIDE PLEASE SO 520 00:27:35,813 --> 00:27:37,448 THIS IS THE CELL LINE, I MEAN 521 00:27:37,448 --> 00:27:40,218 YOU ESTABLISH IT ON 50 OR 60 522 00:27:40,218 --> 00:27:43,154 YEARS AGO, WHAT ABOUT JUST KNOCK 523 00:27:43,154 --> 00:27:46,924 OUT A CELL MHL1 FROM NORMAL OR 524 00:27:46,924 --> 00:27:48,826 NOT A NORMAL MISMATCH REPAIR 525 00:27:48,826 --> 00:27:50,194 PROFICIENT CELLS AND WE ACTUALLY 526 00:27:50,194 --> 00:27:53,464 DID THAT, AS CAN YOU SEE HERE 527 00:27:53,464 --> 00:28:00,338 WHEN YIEW KNOCK OUT MHR1 WHERE 528 00:28:00,338 --> 00:28:03,708 MIDDLE THEN, YOU CAN SEE THE 529 00:28:03,708 --> 00:28:08,980 DEFICIENCY CELLS SHOWING 530 00:28:08,980 --> 00:28:10,047 CYTOSOLIC POINTED BY THE 531 00:28:10,047 --> 00:28:11,215 [INDISCERNIBLE] AND YOU PUT THE 532 00:28:11,215 --> 00:28:14,652 GENES BACK ON THE CELL LINES, 533 00:28:14,652 --> 00:28:18,322 YOU RESTORED THE WILD-TYPE 534 00:28:18,322 --> 00:28:18,623 PHENOMENON. 535 00:28:18,623 --> 00:28:20,758 AND IT'S--WHEN YOU TREAT IT, 536 00:28:20,758 --> 00:28:24,228 TRADE A CELL WITH IONIZING 537 00:28:24,228 --> 00:28:26,964 RADIATION, WITH THE DNA 538 00:28:26,964 --> 00:28:34,772 PRODUCTION, I SHOW IT IN MH6 539 00:28:34,772 --> 00:28:39,844 CELLS SO THIS EVALUATING THE 540 00:28:39,844 --> 00:28:41,813 NEXT SLIDE, PLEASE SO NOW WE 541 00:28:41,813 --> 00:28:44,549 WANT TO SAY WEATHER OR NOT, 542 00:28:44,549 --> 00:28:52,390 DETISHT CELL CAN IMENERATE A 543 00:28:52,390 --> 00:29:02,767 CCAMP AND IN UNTREATED 544 00:29:05,503 --> 00:29:07,972 AND--INCREASED CCAMP PRODUCTION, 545 00:29:07,972 --> 00:29:09,674 MHL1 DEFICIENT CELLS WHICH YOU 546 00:29:09,674 --> 00:29:13,077 CAN SEE UNDER THE LIGHT THERE 547 00:29:13,077 --> 00:29:15,413 FOR QUANTIFICATION, AND SO 548 00:29:15,413 --> 00:29:16,581 SUGGESTING THAT THIS IS 549 00:29:16,581 --> 00:29:26,357 ACTIVATED AND PRODUCED THE CCAMP 550 00:29:26,357 --> 00:29:27,058 NEXT SLIDES. 551 00:29:27,058 --> 00:29:33,264 WE SHOWED YOU, WE CAN CLEARLY 552 00:29:33,264 --> 00:29:40,104 SHOW THAT THIS IS PHOSPHORYLATED 553 00:29:40,104 --> 00:29:42,173 AND THEN IT IS ATTENUATE BUT 554 00:29:42,173 --> 00:29:44,575 BECAUSE OF TIME, I WILL SKIP 555 00:29:44,575 --> 00:29:45,476 THAT BECAUSE IT'S PUBLISHED 556 00:29:45,476 --> 00:29:47,378 ALREADY, IF YOU ARE INTERESTED 557 00:29:47,378 --> 00:29:49,547 YOU CAN LOOK AT THOSE 2 PAPERS. 558 00:29:49,547 --> 00:29:51,549 SO NEXT QUESTION, WE WANT TO ASK 559 00:29:51,549 --> 00:29:53,217 IS PRETTY TOUGH. 560 00:29:53,217 --> 00:29:57,989 SO HOW DOES MHL1 DEFICIENCY CAN 561 00:29:57,989 --> 00:30:00,691 INTRODUCE CYTOSOLIC DNA AND THEN 562 00:30:00,691 --> 00:30:06,464 THIS IS HAS COME TO THE 1 I 563 00:30:06,464 --> 00:30:09,667 SHOWED YOU EARLIER, RECONSITUTE 564 00:30:09,667 --> 00:30:11,469 MISMATCH REPAIR, PAPER WE 565 00:30:11,469 --> 00:30:13,704 PUBLISHED IN 2005. 566 00:30:13,704 --> 00:30:16,474 WE KNOW THAT MLH1 CAN ISHT ACT 567 00:30:16,474 --> 00:30:20,144 WITH EXOMUSEUM CLE ACE 1 AND CAN 568 00:30:20,144 --> 00:30:21,445 NEGATIVELY REGULATE EXO1 MUSEUM 569 00:30:21,445 --> 00:30:24,782 CLE ACE ACTIVITY, SO WIEW KNEW 570 00:30:24,782 --> 00:30:26,083 THIS REGULATION IS CRITICAL FOR 571 00:30:26,083 --> 00:30:32,423 WHETHER OR NOT THE CELL CAN 572 00:30:32,423 --> 00:30:33,524 GENERATE THE CYTOSOLIC DNA. 573 00:30:33,524 --> 00:30:34,792 NEXT SLIDE, PLEASE EMPLOY SO 574 00:30:34,792 --> 00:30:36,327 THEN WE THINK IF THAT'S THE 575 00:30:36,327 --> 00:30:38,796 CASE, IF WE ARE KNOCKING OUT 576 00:30:38,796 --> 00:30:39,530 CERTAINLY--CERTAINLY XO1, SO YOU 577 00:30:39,530 --> 00:30:42,833 MAY NOT BE ABLE TO STAY IN THE 578 00:30:42,833 --> 00:30:44,569 DNA IN THE EMERNG DEFICIENT 579 00:30:44,569 --> 00:30:51,776 CELLS AND THAT'S EXCITING WHAT A 580 00:30:51,776 --> 00:30:56,480 WAY WE SAW AS YOU CAN SEE HERE, 581 00:30:56,480 --> 00:31:00,251 IN THE KNOCK OUT ITS OR IMOWBL 582 00:31:00,251 --> 00:31:01,786 DOCK OUT, MLH1 AND IMOWBL DOCK 583 00:31:01,786 --> 00:31:04,088 OUT AS SHOWN ON THE RIGHT, YOU 584 00:31:04,088 --> 00:31:08,225 REALLY CANNOT SEE THE CYTOSOLIC 585 00:31:08,225 --> 00:31:10,061 DNA AND AGAIN, ARRESTING SHOWS 586 00:31:10,061 --> 00:31:13,030 THAT YOU CAN SAY IMMEDIATELY 587 00:31:13,030 --> 00:31:15,566 THEY ARE ACTIVATION OF CGAS 588 00:31:15,566 --> 00:31:17,301 IMPOSSIBLY WHEN YOU HAVE MLH1 589 00:31:17,301 --> 00:31:20,838 KNOCK OUT BUT A DOUBLE KNOCK 590 00:31:20,838 --> 00:31:24,442 OUT, YOU KNOW PHOSPHORYLATED, SO 591 00:31:24,442 --> 00:31:25,476 SUGGIESTING THAT ACTIVATION FOR 592 00:31:25,476 --> 00:31:28,079 THE INTERNET, IN THE SECOND 593 00:31:28,079 --> 00:31:31,082 PATHWAY, NO MAN CAN EXIST WHEN 594 00:31:31,082 --> 00:31:35,086 YOU KNOCK OUT [INDISCERNIBLE] 1. 595 00:31:35,086 --> 00:31:36,253 NEXT SLIDE, PLEASE. 596 00:31:36,253 --> 00:31:40,491 SO FROM LITERATURE, WE ALSO KNOW 597 00:31:40,491 --> 00:31:43,461 THAT MLH1 IS INVOLVED IN TOP O 598 00:31:43,461 --> 00:31:44,996 STRAND BREAK REPAIR AS SHOWN IN 599 00:31:44,996 --> 00:31:54,271 THIS PAPER, NEXT SLIDE, PLEASE. 600 00:31:54,271 --> 00:31:57,375 SO WE BELIEVE THAT MLH1 IS 601 00:31:57,375 --> 00:32:00,144 CONTROL ACTIVITY IN THE DOUBLE 602 00:32:00,144 --> 00:32:01,846 STRANDED BREAK REPAIR WITH THE 603 00:32:01,846 --> 00:32:02,780 FOUNDATION, AND MISMATCH REPAIR 604 00:32:02,780 --> 00:32:05,316 AS I SHOW YOU EARLIER. 605 00:32:05,316 --> 00:32:12,456 SO IN DOUBLE STRANDED REPAIR, IN 606 00:32:12,456 --> 00:32:14,825 THE RESECTION, AND ACTUALLY 1 OF 607 00:32:14,825 --> 00:32:17,662 THE ENZYME I HAD BEEN HOEING IN 608 00:32:17,662 --> 00:32:20,531 THIS ENDORESECTION, AND WE 609 00:32:20,531 --> 00:32:22,933 BELIEVE MLH1 OR MUTANT L-ALPHA 610 00:32:22,933 --> 00:32:25,002 ALSO CONTROLS THIS PROCESS, IF 611 00:32:25,002 --> 00:32:28,472 YOU DON'T HAVE MUTANT LR-5 SHOWN 612 00:32:28,472 --> 00:32:30,341 IN THE RIGHT, THE EXCISION WILL 613 00:32:30,341 --> 00:32:33,010 BE REALLY EXTENSIVE AND GENERATE 614 00:32:33,010 --> 00:32:34,879 A LARGER QUANTITY OF SINGLE 615 00:32:34,879 --> 00:32:39,450 STRANDED DNA WHICH ACCRUED, GAVE 616 00:32:39,450 --> 00:32:40,951 THE CHROMOSOME, BIGGER PROBLEMS, 617 00:32:40,951 --> 00:32:44,655 SO THEY WILL BE CHROMOSOME BREAK 618 00:32:44,655 --> 00:32:44,989 OUTS. 619 00:32:44,989 --> 00:32:45,423 SO NEXT SLIDE. 620 00:32:45,423 --> 00:32:46,991 SO WE WANT TO SAY WHETHER OR NOT 621 00:32:46,991 --> 00:32:49,960 IF THAT'S REALLY THE CASE 622 00:32:49,960 --> 00:32:54,765 BECAUSE MLH1 AND EXO1 INTACT, 623 00:32:54,765 --> 00:32:57,735 WITH EACH OTHER AND THE DOMAIN 624 00:32:57,735 --> 00:33:02,873 HAS BEEN AMENDED BY SEVERAL 625 00:33:02,873 --> 00:33:03,474 LABORATORY, PARTICULARLY 626 00:33:03,474 --> 00:33:04,475 [INDISCERNIBLE] LABORATORY 627 00:33:04,475 --> 00:33:07,945 SHOWING THEY ARE INTO 628 00:33:07,945 --> 00:33:10,381 INTERACTIVE DOMAINS, MLH1, AND 629 00:33:10,381 --> 00:33:13,684 EXO1, ARE SHARED BY THIS GREEN 630 00:33:13,684 --> 00:33:14,452 AREA. 631 00:33:14,452 --> 00:33:18,222 SO WE TRY TO MARK OUT THIS 632 00:33:18,222 --> 00:33:22,727 INDUCTION DOMAIN AND WE TAKE 633 00:33:22,727 --> 00:33:24,929 EITHER TAKING THE FF, WHICH IS 634 00:33:24,929 --> 00:33:29,600 THE MIDDLE ACTION, OR TAKING OUT 635 00:33:29,600 --> 00:33:31,769 THE WHOLE C-TERMINAL OR COMBINE 636 00:33:31,769 --> 00:33:34,138 THIS 2 TOGETHER, SO I SHOWED 637 00:33:34,138 --> 00:33:35,840 HERE, THE PURIFIED PROTEIN IN 638 00:33:35,840 --> 00:33:38,275 THE BOTTOM LEFT AND THEN THERE 639 00:33:38,275 --> 00:33:40,377 ARE 3 MUTANT, I SHOWED HERE AND 640 00:33:40,377 --> 00:33:43,013 THEN ON THE RIGHT, BOTTOM RIGHT, 641 00:33:43,013 --> 00:33:45,816 AND THIS IS ACTUALLY 642 00:33:45,816 --> 00:33:46,917 SHOWING--TRIED TO SHOW 643 00:33:46,917 --> 00:33:49,720 INTERACTION OF THIS 3 MUTANT 644 00:33:49,720 --> 00:33:50,955 PROTEIN WITH MUTANT L-ALPHA AS 645 00:33:50,955 --> 00:33:54,458 YOU CAN SEE IN THE FAR RIGHT, 646 00:33:54,458 --> 00:33:56,961 THE DOWNL MUTANT, WHERE WE 647 00:33:56,961 --> 00:34:00,631 KNOCKING OUT THE C-TERMINAL AND 648 00:34:00,631 --> 00:34:03,968 ALSO THE FF [INDISCERNIBLE] 649 00:34:03,968 --> 00:34:08,405 AA YOU DO REALLY SEE THE 650 00:34:08,405 --> 00:34:10,141 INTERACTION BETWEEN MUTANT 651 00:34:10,141 --> 00:34:11,575 L-ALPHA AND EXO1 AND THAT'S 652 00:34:11,575 --> 00:34:13,911 EXACTLY WHAT WE NEED AND WE KNOW 653 00:34:13,911 --> 00:34:17,114 WE ACTUALLY TASK THIS MUTANT, 654 00:34:17,114 --> 00:34:20,351 ALL THIS MUTANT HAVE THE REALLY 655 00:34:20,351 --> 00:34:23,187 ACTIVE NUCLEACE ACTIVITY, SO 1 656 00:34:23,187 --> 00:34:28,692 PAST WITH THIS MUTANT 657 00:34:28,692 --> 00:34:29,527 ENDORESECTION, CATALYZE THE 658 00:34:29,527 --> 00:34:36,467 ENDORESECTION CAN BE CONTROLLED 659 00:34:36,467 --> 00:34:37,234 BY THE ALPHA. 660 00:34:37,234 --> 00:34:38,769 NEXT SLIDE, PLEASE. 661 00:34:38,769 --> 00:34:44,241 SO THIS IS THE ENDOASSAY 662 00:34:44,241 --> 00:34:47,011 NUCLEUS, THIS CREATED BY 663 00:34:47,011 --> 00:34:48,779 DR. [INDISCERNIBLE] DIFFICULT TR 664 00:34:48,779 --> 00:34:55,486 ME TO SAY LAST NIEM, I MEAN TRY 665 00:34:55,486 --> 00:34:59,323 TO USE DUKE [INDISCERNIBLE] NAME 666 00:34:59,323 --> 00:35:01,492 ALTHOUGH THEY ARE NOT EXACTLY 667 00:35:01,492 --> 00:35:03,494 THE SAME NAME, SO AS YOU CAN 668 00:35:03,494 --> 00:35:11,235 SEE, HERE, ON THE RIGHT, WE ARE 669 00:35:11,235 --> 00:35:12,970 USING THE WILD-TYPE--EXO1, AND 670 00:35:12,970 --> 00:35:14,839 THE MUTANT EXO1 TO DO THIS 671 00:35:14,839 --> 00:35:16,941 ENDORESECTION IN THE PRESENCE OR 672 00:35:16,941 --> 00:35:19,810 ABSENCE OF MUTANT L-ALPHA SO 673 00:35:19,810 --> 00:35:22,213 WHEN YOU SAY, WILL YOU USE 674 00:35:22,213 --> 00:35:26,250 WILD-TYPE AXON 1 IN THE ABSENCE 675 00:35:26,250 --> 00:35:29,320 OF MUTANT LH1, CAN YOU ONLY SEE 676 00:35:29,320 --> 00:35:32,323 THE PRODUCT OF VERY SHORT, WE 677 00:35:32,323 --> 00:35:34,758 NAME THE PRODUCT 1 AND THEN WHEN 678 00:35:34,758 --> 00:35:37,428 YOU INCREASE THE MUTANT-L 679 00:35:37,428 --> 00:35:42,600 CONCENTRATION IN THIS REACTION, 680 00:35:42,600 --> 00:35:44,134 CAN YOU SEE THE [INDISCERNIBLE] 681 00:35:44,134 --> 00:35:46,237 COMING BACK, SO MUCH BIGGER SO 682 00:35:46,237 --> 00:35:48,105 SUGGESTING IT CONTROLS THE 683 00:35:48,105 --> 00:35:49,073 NUCLEUS ACTIVITY OF 1, HOWEVER, 684 00:35:49,073 --> 00:35:52,610 IF YOU LOOK AT THE FAR RIGHT 1 685 00:35:52,610 --> 00:35:55,145 WHICH WE SHOWED THIS 1 LITTLE IN 686 00:35:55,145 --> 00:35:57,248 THE ACTION, CONTAIN THIS MUTANT 687 00:35:57,248 --> 00:35:59,350 X-1 AND THE MUTANT L-ALPHA WHAT 688 00:35:59,350 --> 00:36:01,485 DO YOU SEE, YOU REALLY CAN HAVE 689 00:36:01,485 --> 00:36:04,488 THE SEE MUCH OF THIS WITH 2 IN 690 00:36:04,488 --> 00:36:09,460 THIS REACTION, SO SUGGESTING 691 00:36:09,460 --> 00:36:12,196 EXTENSIVE ECCISIONS IN THIS 692 00:36:12,196 --> 00:36:14,398 MUTANT REACTION SO SUGGESTING, 693 00:36:14,398 --> 00:36:16,734 GENERATING A LOT OF SINGLE 694 00:36:16,734 --> 00:36:20,704 STRANDED DNA, NEXT SLIDE, 695 00:36:20,704 --> 00:36:20,938 PLEASE. 696 00:36:20,938 --> 00:36:23,807 SO WHEN WE LOOK AT THIS, WHETHER 697 00:36:23,807 --> 00:36:25,943 OR NOT THAT--THEY'RE NOT SINGLE 698 00:36:25,943 --> 00:36:29,013 STRAND DNA GENERATED AND AS YOU 699 00:36:29,013 --> 00:36:33,017 CAN SEE HERE, IN MLH1 DEFICIENT 700 00:36:33,017 --> 00:36:38,355 CELLS AND WE DO THIS BRDU 701 00:36:38,355 --> 00:36:45,696 INCORPORATION AND YOU CAN SEE A 702 00:36:45,696 --> 00:36:49,700 LOT OF BRDU INCORPORATED IN MLH1 703 00:36:49,700 --> 00:36:52,136 CELLS COMPARED WITH WILD-TYPE 704 00:36:52,136 --> 00:36:55,272 AND ALSO IN THE RPA BINDING, YOU 705 00:36:55,272 --> 00:36:58,642 CAN SEE NICELY, MHL1 DEFICIENT 706 00:36:58,642 --> 00:37:01,011 CELLS BUT NOT MANY IN WILD-TYPE 707 00:37:01,011 --> 00:37:02,146 CELLS, SO THAT'S--I WAS THINKING 708 00:37:02,146 --> 00:37:12,489 WHAT DO WE THINK IS HAPPENING IN 709 00:37:12,489 --> 00:37:15,693 THERE IN VIVO IN THE SUCCESSES 710 00:37:15,693 --> 00:37:17,528 SECTION PLEASE. 711 00:37:17,528 --> 00:37:18,362 THIS WAS CHROMOSOME 712 00:37:18,362 --> 00:37:22,533 ABNORMALITIES SO WE DID THIS 713 00:37:22,533 --> 00:37:27,037 EXPERIMENT, CAN YOU SEE MLH1 714 00:37:27,037 --> 00:37:29,340 DEFESHT CELLS AND ALSO IN HUMAN 715 00:37:29,340 --> 00:37:31,308 CELLS WE SHOW EXACTLY THE SAME. 716 00:37:31,308 --> 00:37:41,852 THEY'RE NOT ALL [INDISCERNIBLE] 717 00:38:03,774 --> 00:38:05,843 IN THE SO WE CAN'T SO HOW THESE 718 00:38:05,843 --> 00:38:07,644 ARE AWARDED TO 2, THE DNA BY THE 719 00:38:07,644 --> 00:38:10,414 WAY WE BELIEVE, THAT'S PROBABLY 720 00:38:10,414 --> 00:38:11,715 HAPPENING AND THAT'S WHERE I'M 721 00:38:11,715 --> 00:38:13,283 LOOKING AT IT, BECAUSE I YOU CAN 722 00:38:13,283 --> 00:38:15,486 IMAGINE HOW IT OCCURRED SO THE 723 00:38:15,486 --> 00:38:20,290 MODEL, BASED ON THE DATA I SHOW 724 00:38:20,290 --> 00:38:22,359 YOU, I CAN--I CALCULATE IT HERE 725 00:38:22,359 --> 00:38:25,229 IS THAT, ON THE LEFT SIDE, WHERE 726 00:38:25,229 --> 00:38:27,898 YOU HAVE THE MUTANT L-PROTEIN 727 00:38:27,898 --> 00:38:30,768 HERE, SO YOU ARE ENDORESECTION, 728 00:38:30,768 --> 00:38:41,245 IN DOUBLE STRANDED BREAK, IS 729 00:38:50,921 --> 00:38:52,389 REALLY LIMITED ON THE RIGHT, AND 730 00:38:52,389 --> 00:38:54,591 IF YOU HAVE LIMITED PROTEIN IN 731 00:38:54,591 --> 00:38:58,529 THE CELL, OR BREAK FOR X-1 732 00:38:58,529 --> 00:38:59,563 NUCLEUS ACTIVITY, SO THE 733 00:38:59,563 --> 00:39:01,231 DECISION CAN BE REALLY 734 00:39:01,231 --> 00:39:03,967 EXTENSIVE, GENERALLY A LOT OF 735 00:39:03,967 --> 00:39:05,969 SINGLE STRAND DNA, AND YOU CAN 736 00:39:05,969 --> 00:39:08,605 SOURCE THE RPA SO YOU WANT 737 00:39:08,605 --> 00:39:10,240 SINGLE STRAND DNA CANNOT BE 738 00:39:10,240 --> 00:39:11,842 PROTECTED SO THAT'S WHY YOU 739 00:39:11,842 --> 00:39:13,944 CAUSE THE CHROMOSOME BREAK AND 740 00:39:13,944 --> 00:39:18,182 YOU SET IT OF THE DNA WHICH CAN 741 00:39:18,182 --> 00:39:23,620 INDUCE THE C-GAS DEACTIVATION 742 00:39:23,620 --> 00:39:25,322 WHICH FACILITATED, AND IT'S THE 743 00:39:25,322 --> 00:39:31,695 NATION FOR YOU FOR 50%. 744 00:39:31,695 --> 00:39:34,198 FOR THE DEFICIENT 1S TO RESPOND 745 00:39:34,198 --> 00:39:38,102 AND WE BELIEVE THAT CGAS IS 746 00:39:38,102 --> 00:39:39,069 IMPOSSIBLE, PROBABLY 747 00:39:39,069 --> 00:39:44,108 CONTRIBUTING TO THOSE SOME OF 748 00:39:44,108 --> 00:39:46,009 THOSE BECAUSE MISMATCH REPAIR 749 00:39:46,009 --> 00:39:49,780 DEFICIENCY CAN CAUSE ANY GENE 750 00:39:49,780 --> 00:39:51,615 MUTATIONS, SO CGAS IS CAN THEN 751 00:39:51,615 --> 00:39:53,550 PASS AWAY, ALL THE YEEPS CAN BE 752 00:39:53,550 --> 00:39:56,420 MUTATED IF THAT HAPPENING, AND 753 00:39:56,420 --> 00:39:58,856 THEN, THOSE TUMORS EVEN THEY ARE 754 00:39:58,856 --> 00:39:59,723 DEFECTIVE IN MISMATCH REPAIR, 755 00:39:59,723 --> 00:40:02,192 THEY'RE NOT GOING TO RESPOND FOR 756 00:40:02,192 --> 00:40:03,694 THE IMMUNOTHERAPY, SO THAT'S WHY 757 00:40:03,694 --> 00:40:09,733 I PUT THAT MARKER HERE, SO NEXT 758 00:40:09,733 --> 00:40:14,338 SLIDE, PLEASE TO, EVALUATED THAT 759 00:40:14,338 --> 00:40:24,781 SO WE, COME BACK, CONTACT 760 00:40:26,183 --> 00:40:29,453 FOR--COME ON THERE OF THE WHO 761 00:40:29,453 --> 00:40:33,090 INITIALLY SHOWED THE MISMATCH 762 00:40:33,090 --> 00:40:35,559 REPAIR DEFICIENCY, HELP THE 763 00:40:35,559 --> 00:40:35,893 IMMUNOTHERAPY. 764 00:40:35,893 --> 00:40:38,795 SO WE HAVE TO HAVE PATIENTS 765 00:40:38,795 --> 00:40:40,764 DEFECTIVE, WITH THE MH1 IS 766 00:40:40,764 --> 00:40:42,332 TREATED WITH THERAPY AND THIS IS 767 00:40:42,332 --> 00:40:46,036 THE PAPER, CAN YOU SEE, SO 768 00:40:46,036 --> 00:40:48,839 WHENEVER THE PATIENT HAVE 769 00:40:48,839 --> 00:40:50,140 REALIZED OR GO TO EXPRESSION OF 770 00:40:50,140 --> 00:40:52,743 CGAS AS SHOWN IN THE LEFT OR 771 00:40:52,743 --> 00:40:57,314 STING ON THE RIGHT, THEN THEY 772 00:40:57,314 --> 00:41:00,517 RESPONDED TO THE PDL1 ANTIBODY 773 00:41:00,517 --> 00:41:02,019 TREATMENT AS WELL, BUT IF THEY 774 00:41:02,019 --> 00:41:03,987 DON'T HAVE THIS GOOD EXPRESSION, 775 00:41:03,987 --> 00:41:07,891 THOSE PATIENT DO NOT RESPOND TO 776 00:41:07,891 --> 00:41:17,501 IMMUNOTHERAPY AT ALL, SO THIS 777 00:41:17,501 --> 00:41:19,136 CONFORMS OUR DATA 778 00:41:19,136 --> 00:41:21,004 [INDISCERNIBLE] FOR BIOCHEMICAL 779 00:41:21,004 --> 00:41:21,338 ASSAYS. 780 00:41:21,338 --> 00:41:22,573 NEXT SLIDE, PLEASE, SO I HOPE I 781 00:41:22,573 --> 00:41:26,276 HAVE SHOWED YOU THAT WE HAVE 782 00:41:26,276 --> 00:41:30,380 ACTUALLY FOUND THE MLH1 783 00:41:30,380 --> 00:41:34,351 DEFICIENCY, AND CGAS, AND THE 784 00:41:34,351 --> 00:41:38,422 DEFICIENT OR NO EXPRESSION OF 785 00:41:38,422 --> 00:41:42,159 CGAS THIS FACTOR CAN BE CAUSED 786 00:41:42,159 --> 00:41:44,995 BY MISMATCH REPAIR DEFICIENCY 787 00:41:44,995 --> 00:41:49,199 CONTRIBUTES TO THE 50% OR MAYBE 788 00:41:49,199 --> 00:41:53,737 CERTAIN IN THIS MISMATCH REPAIR 789 00:41:53,737 --> 00:41:56,173 ALLOW THEM TO EXIST WITH THE 790 00:41:56,173 --> 00:42:03,046 IMMUNOTHERAPY AND WE ALSO SHOW 791 00:42:03,046 --> 00:42:05,115 THAT ACTUALLY REALLY IMPORTANT 792 00:42:05,115 --> 00:42:11,421 BUT CONTROLLING EXO1 ACTIVITY. 793 00:42:11,421 --> 00:42:12,256 NEXT SLIDE, PLEASE. 794 00:42:12,256 --> 00:42:14,424 SO NOWIME GOING TO SWITCH, AND 795 00:42:14,424 --> 00:42:18,895 IT'S TALKING ABOUT THIS MLH1 796 00:42:18,895 --> 00:42:21,265 MUTANT L-ALPHA CONTROL, THE EXO1 797 00:42:21,265 --> 00:42:24,434 NUCLEACE ACTIVITY AND WE SHOW 798 00:42:24,434 --> 00:42:34,878 THIS ACTUAL LOW, ATHE SAME 799 00:42:40,951 --> 00:42:43,420 FUNCTION, SEEMS DOING AND 800 00:42:43,420 --> 00:42:44,655 CONTRIBUTING TO EXO1 GENERATING. 801 00:42:44,655 --> 00:42:46,890 SO NEXT SLIDE PLEASE. 802 00:42:46,890 --> 00:42:50,961 SO I SHOWED OTHER IN THE 803 00:42:50,961 --> 00:42:52,462 AUDIENCE, ALSO THE 804 00:42:52,462 --> 00:42:55,165 [INDISCERNIBLE] AND THE 805 00:42:55,165 --> 00:42:55,532 [INDISCERNIBLE]. 806 00:42:55,532 --> 00:43:06,009 SO AS YOU PROBABLY KNOW, YOU 807 00:43:07,311 --> 00:43:09,212 HADITANT SYSTEM ACTUALLY BRAIN 808 00:43:09,212 --> 00:43:13,350 DISORDER AND IT'S A GENE AND OUR 809 00:43:13,350 --> 00:43:15,085 NORMAL INDIVIDUAL, YOU WOULD 810 00:43:15,085 --> 00:43:18,255 HAVE REPEAT NUMBER AROUND THE 811 00:43:18,255 --> 00:43:20,991 [INDISCERNIBLE] BUT ONCE THIS 812 00:43:20,991 --> 00:43:23,093 CAKB SIN REPEATS, IT REACHES 36, 813 00:43:23,093 --> 00:43:28,398 THEN YOU WILL HAVE ISSUES AND 814 00:43:28,398 --> 00:43:31,368 THERE ARE PATIENTS, YOU KNOW 815 00:43:31,368 --> 00:43:36,540 THAT, SO THIS EXPRESSION WILL 816 00:43:36,540 --> 00:43:38,775 ALLOW THE WILD-TYPE HTT TO 817 00:43:38,775 --> 00:43:43,113 MUTATE TO THE FORM OF HTT, AND 818 00:43:43,113 --> 00:43:45,215 THE SO CALLED MHTT. 819 00:43:45,215 --> 00:43:47,284 OKAY, THIS PROTEIN, THIS 820 00:43:47,284 --> 00:43:48,785 MUTATING THERE, WITH THE 821 00:43:48,785 --> 00:43:53,156 PROTEIN, HAS BEEN SHARED TO 822 00:43:53,156 --> 00:43:54,725 INDUCE CGAS ACTIVATION, MEDIATED 823 00:43:54,725 --> 00:43:56,660 APOPTOSIS BUT THE NECKANISM IS 824 00:43:56,660 --> 00:43:59,863 NOT REALLY LOW, AND WE KNOW FROM 825 00:43:59,863 --> 00:44:03,800 PREVIOUS TODAY, THAT MISMATCHED 826 00:44:03,800 --> 00:44:07,537 REPAIR GENES, PARTICULARLY MSH3 827 00:44:07,537 --> 00:44:09,539 AND MHSL3, ARE KEY GENETIC 828 00:44:09,539 --> 00:44:11,174 MODIFIERS BUT NOW PEOPLE ARE 829 00:44:11,174 --> 00:44:12,709 THINKING NOT JUST THE MODIFIER, 830 00:44:12,709 --> 00:44:16,413 IT'S JUST THE DRIVERS OF HUNTING 831 00:44:16,413 --> 00:44:23,019 TON'S DEC. 832 00:44:23,019 --> 00:44:23,520 NEXT SLIDE, PLEASE. 833 00:44:23,520 --> 00:44:28,091 SO WE WILL TRY TO--I MEAN, MLH3, 834 00:44:28,091 --> 00:44:30,093 WHICH IS THE SUBUNIT OF THE 835 00:44:30,093 --> 00:44:33,430 MUTANTS ALPHA THERE ARE MANY 836 00:44:33,430 --> 00:44:35,499 LABORATORIES SHOWING THAT THE 837 00:44:35,499 --> 00:44:37,834 MUTANT ALPHA, HOW MUTANTS ALPHA 838 00:44:37,834 --> 00:44:39,936 CONTRIBUTED TO REPEAT THE 839 00:44:39,936 --> 00:44:42,172 EXPANSION AND RELATED TO HUNTING 840 00:44:42,172 --> 00:44:48,712 TON'S DEC AND ENCLOUDING 841 00:44:48,712 --> 00:44:51,715 DR. [INDISCERNIBLE] SHE WAS IN A 842 00:44:51,715 --> 00:44:53,950 MEETING, SHOWING HOW--I MEAN 843 00:44:53,950 --> 00:44:56,920 MUTANT S-BETA IS BINDING AND 844 00:44:56,920 --> 00:44:58,455 PROMOTE EXPANSIONS, THERE ARE 845 00:44:58,455 --> 00:44:59,623 MANY OTHER LABORATORIES ALSO 846 00:44:59,623 --> 00:45:03,393 WORK ON THAT, HOWEVER, HOW MLH1 847 00:45:03,393 --> 00:45:06,596 CONTRIBUTED TO THIS IS NOT 848 00:45:06,596 --> 00:45:08,064 REALLY KNOW, NOWHERE THE 849 00:45:08,064 --> 00:45:09,032 MECHANISM OUT OF THERE. 850 00:45:09,032 --> 00:45:11,935 SO WE HAVE A 2 POSTER FELLOWS 851 00:45:11,935 --> 00:45:13,470 WHILE WE'RE INTERESTED IN THIS 852 00:45:13,470 --> 00:45:17,574 PROJECT, AND TRIED LOOKING AT 853 00:45:17,574 --> 00:45:20,777 HOW MLH1 DOING AND THEY BELIEVE 854 00:45:20,777 --> 00:45:22,379 THAT MLH1 PROBABLY IPT GREATER 855 00:45:22,379 --> 00:45:23,914 TACT WITH THE HUNTING TON 856 00:45:23,914 --> 00:45:29,052 PROTEIN, NEXT SLIDE, PLEASE. 857 00:45:29,052 --> 00:45:31,955 AND WITH THAT IN MIND, THEY 858 00:45:31,955 --> 00:45:33,223 ACTUALLY TRY TO DETERMINE 859 00:45:33,223 --> 00:45:36,126 WHETHER OR NOT MUTANT L-PROTEIN 860 00:45:36,126 --> 00:45:39,162 IMPACT WITH HUNTING TON PROTEIN, 861 00:45:39,162 --> 00:45:43,467 AND THEY DID WITH PURIFIED 862 00:45:43,467 --> 00:45:44,601 PROTEIN, WITH IMMUNO 863 00:45:44,601 --> 00:45:46,336 PRECIPITATION, WERE ALSO USING 864 00:45:46,336 --> 00:45:48,638 THE CELL, I MEAN THEY JUST 865 00:45:48,638 --> 00:45:50,474 [INDISCERNIBLE], THE MUTANT 866 00:45:50,474 --> 00:45:51,775 L-PROTEIN AND DIFFERENT LENGTH 867 00:45:51,775 --> 00:45:53,743 OF HUNTING TON DEC OR REPEATS 868 00:45:53,743 --> 00:45:58,448 HUNTING TON PROTEIN IN CELL, IT 869 00:45:58,448 --> 00:46:01,051 SHOWS REAL NICELY, SO THE 870 00:46:01,051 --> 00:46:03,220 WILD-TYPE HUNTING TON PROTEIN 871 00:46:03,220 --> 00:46:06,056 SHOWING IN THE WESTERN BLOT CAN 872 00:46:06,056 --> 00:46:10,694 INTERACT WITH BOTH EXO1 AND 873 00:46:10,694 --> 00:46:11,094 MUTANT L-ALPHA. 874 00:46:11,094 --> 00:46:14,498 BUT NOT THE MUTATED HGT, SO BOTH 875 00:46:14,498 --> 00:46:18,702 SHOWED THE SAME RESULT, AND 876 00:46:18,702 --> 00:46:21,171 WE'RE ALSO USING THE PROXIMITY 877 00:46:21,171 --> 00:46:22,439 MEDICATION ASSAY, SHOWN ON THE 878 00:46:22,439 --> 00:46:25,976 RIGHT, YOU CAN SEE THAT THE 879 00:46:25,976 --> 00:46:32,582 SHORTER 1, I MEAN, IF HTT ONLY 880 00:46:32,582 --> 00:46:35,352 CONTAIN 23 CUE, I MEAN SAGE, 881 00:46:35,352 --> 00:46:40,757 REPEAT, AND THEN THEY CAN BUT, 882 00:46:40,757 --> 00:46:44,060 IF YOU INCREASE THIS REPEATS TO 883 00:46:44,060 --> 00:46:47,130 73, AND THE INDUCTION 884 00:46:47,130 --> 00:46:50,667 [INDISCERNIBLE] SO THIS DATA IS 885 00:46:50,667 --> 00:46:51,935 REALLY CONSISTENT WITH OUR 886 00:46:51,935 --> 00:46:56,606 WESTERN BLOT OF DATA, IMMUNO 887 00:46:56,606 --> 00:46:59,643 PRECIPITATION DATA. 888 00:46:59,643 --> 00:47:00,110 NEXT SLIDE, PLEASE. 889 00:47:00,110 --> 00:47:05,315 NEXT WE WANTED TO KNOW WHERE 890 00:47:05,315 --> 00:47:07,751 THIS HTT IMPACTS WITH EXO1, SO 891 00:47:07,751 --> 00:47:12,789 WE CHOP DOWN THE X-WHY INTO 892 00:47:12,789 --> 00:47:20,730 MODIFIED FRAGMENTS AND YOU CAN 893 00:47:20,730 --> 00:47:22,732 SEE THEM AND THEY'RE REGULATING 894 00:47:22,732 --> 00:47:27,270 HOW INTACT WITH HTT AND YOU CAN 895 00:47:27,270 --> 00:47:31,474 SEE ON THE RIGHT AND THEN, WE 896 00:47:31,474 --> 00:47:33,944 HAVE THIS NUMBER OF THE SAME AS 897 00:47:33,944 --> 00:47:38,114 THE FLASH NUMBER ON THE LEFT, 898 00:47:38,114 --> 00:47:42,152 AND FRACTION OR FRAGMENT 1-3, 899 00:47:42,152 --> 00:47:45,322 1-4 AND ALSO SEVERAL AND YOU CAN 900 00:47:45,322 --> 00:47:51,261 SEE SEVERAL ON THE LAB AND IT'S 901 00:47:51,261 --> 00:47:54,764 A FRAGMENT IN BLUE, INTACT WITH 902 00:47:54,764 --> 00:47:56,399 PROTEIN, VERY, VERY WELL, THAT'S 903 00:47:56,399 --> 00:47:57,801 PROBABLY THE BEST, AND YOU HAVE 904 00:47:57,801 --> 00:48:00,303 THAT AND THEN THE FULL 905 00:48:00,303 --> 00:48:00,670 [INDISCERNIBLE]. 906 00:48:00,670 --> 00:48:05,175 SO WE BELIEVE THIS IS--THIS IS 907 00:48:05,175 --> 00:48:09,512 THE REASON THE REASON HOW 908 00:48:09,512 --> 00:48:16,653 HUNTING TON PROTEIN SOMEHOW ALSO 909 00:48:16,653 --> 00:48:19,422 REGULATE THE EXO1 ACTIVITY, AND 910 00:48:19,422 --> 00:48:22,058 INTERESTINGLY WE KNOW THIS 911 00:48:22,058 --> 00:48:23,360 IS--THIS REGION, THIS FRAGMENT 912 00:48:23,360 --> 00:48:28,031 IS SERVING IT IS IN THE NUCLEUS 913 00:48:28,031 --> 00:48:30,433 DOMAIN, SO YOU CLUE CLE ACE 914 00:48:30,433 --> 00:48:32,569 DOMAIN, WE SEE IT FOR MUTANT L 915 00:48:32,569 --> 00:48:34,738 PROTEIN, SO NEXT SLIDE, PLEASE. 916 00:48:34,738 --> 00:48:39,075 NOW WE DO EXACTLY THE SAME 917 00:48:39,075 --> 00:48:41,177 ENDORESECTION ASSAY AS I DID 918 00:48:41,177 --> 00:48:44,614 EARLIER, AND THEN YOU CAN SEE, 919 00:48:44,614 --> 00:48:47,150 EXO1 BY ITSELF CAN AJUST THE DNA 920 00:48:47,150 --> 00:48:54,357 INTO MORE PIECES AND THEN, WHEN 921 00:48:54,357 --> 00:48:57,127 YOU--WHEN YOU INCUBATE EXO1 WITH 922 00:48:57,127 --> 00:49:02,666 THE MUTANT PROTEIN, AND YOU KNOW 923 00:49:02,666 --> 00:49:05,635 NO LONGER SHOW THAT EXO 1 ITSELF 924 00:49:05,635 --> 00:49:09,005 BUT IF YOU INCUBATE THE EXO1 925 00:49:09,005 --> 00:49:13,276 WITH MUTANT L-ALPHA OR WILD-TYPE 926 00:49:13,276 --> 00:49:15,345 HTT, THEN CAN YOU SEE THE 927 00:49:15,345 --> 00:49:19,315 NUCLEACE ACTIVITIES IS REALLY 928 00:49:19,315 --> 00:49:21,985 SUPPRESSED, NOW YOU CUT A LITTLE 929 00:49:21,985 --> 00:49:24,754 BIT, NOT A--ALL THE WAY TO 930 00:49:24,754 --> 00:49:27,691 THERE'S MORE PIECES, OKAY. 931 00:49:27,691 --> 00:49:30,427 AND INTERESTINGLY, IF YOU 932 00:49:30,427 --> 00:49:33,396 INCUBATE MUTANT L-ALPHA AND EXO1 933 00:49:33,396 --> 00:49:37,500 AND THE MUTANT HTT, YOU SEND 934 00:49:37,500 --> 00:49:38,735 MUTANT L-ALPHA 1 BY TOPPING IT. 935 00:49:38,735 --> 00:49:43,406 SO NOW THE QUESTION IS, IF WE 936 00:49:43,406 --> 00:49:46,910 THINK THIS EXO 1, AND MUTANT 937 00:49:46,910 --> 00:49:52,582 THERE PROTEIN CANNOT CONTROL EXO 938 00:49:52,582 --> 00:49:54,451 1'S ACTIVITY,--FIND THERE AND 939 00:49:54,451 --> 00:49:56,052 HOW MUTANT L-ALPHA SHOULD BE 940 00:49:56,052 --> 00:49:58,822 ABLE TO CONTROL THE NUCLEUS 941 00:49:58,822 --> 00:50:00,190 ACTIVITY, SO WHY WE SEE THE 942 00:50:00,190 --> 00:50:05,462 PROBLEM IN THE MUTANT CELLS, SO 943 00:50:05,462 --> 00:50:08,698 NEXT SLIDE. 944 00:50:08,698 --> 00:50:15,538 SO THEN WE SAY--I'M SORRY, THIS 945 00:50:15,538 --> 00:50:16,606 IS SHOWING THE ENDORESECTION 946 00:50:16,606 --> 00:50:19,275 ASSAY, WHERE WE WANT TO SAY, WE 947 00:50:19,275 --> 00:50:22,378 ONLY HAVE 1, 1 DOUBLE STRANDED 948 00:50:22,378 --> 00:50:25,181 BREAK AND WANT TO SEE HOW THE 949 00:50:25,181 --> 00:50:27,283 RESECTION, ENDORESECTION GOES, 950 00:50:27,283 --> 00:50:31,387 WE HAVE 3 DIFFERENT PLACES 951 00:50:31,387 --> 00:50:32,188 CONTAINED THIS RESTRICTION SIZE 952 00:50:32,188 --> 00:50:36,192 AND THEN WE WILL BE ABLE TO 953 00:50:36,192 --> 00:50:38,495 MODULATING AMOUNT OF SINGLE 954 00:50:38,495 --> 00:50:41,164 STRAND D NA GENERATED, AND AS 955 00:50:41,164 --> 00:50:43,466 YOU CAN SEE IN THE BOTTOM THAT 956 00:50:43,466 --> 00:50:49,739 YOU REALLY MUTANT CELLS THAT 957 00:50:49,739 --> 00:50:51,574 HTT CELLS THAT DEGENERATE A LOT 958 00:50:51,574 --> 00:50:55,245 OF SINGLE STRAND DNA HAD IS ON 959 00:50:55,245 --> 00:50:59,783 THE RIGHT, THE RIGHT COLUMN, SO 960 00:50:59,783 --> 00:51:01,951 SUGGESTING THAT WHEN HAVE YOU 961 00:51:01,951 --> 00:51:04,420 THIS MUTATED HTT AND THEY'RE NOT 962 00:51:04,420 --> 00:51:11,494 SINGLE STRANDED DNA GENERATED SO 963 00:51:11,494 --> 00:51:15,265 WHAT HAPPENED TO THIS MUTANT L 964 00:51:15,265 --> 00:51:16,533 PROTEIN, WORSE, MUTANT L GOING 965 00:51:16,533 --> 00:51:19,936 INTO FUNCTION WITHOUT THE 966 00:51:19,936 --> 00:51:24,474 PRESENCE OF MTT, SO NEXT SLIDE, 967 00:51:24,474 --> 00:51:25,909 PLEASE. 968 00:51:25,909 --> 00:51:27,811 SO WE HAD THE IMAGE OF THE HALF 969 00:51:27,811 --> 00:51:32,115 LIFE OF THE MUTANT L-ALPHA 970 00:51:32,115 --> 00:51:33,516 EMPLOY OUR HYPOTHESIS IS THAT 971 00:51:33,516 --> 00:51:36,152 MAYBE THIS MUSEUM TABT PROTEIN 972 00:51:36,152 --> 00:51:37,787 CAN DETROIT MUTANT L-PROTEIN, 973 00:51:37,787 --> 00:51:40,190 THAT'S INDEED WHAT WE FIND, THE 974 00:51:40,190 --> 00:51:42,759 HALF LIFE OF MLH1 IS MUCH 975 00:51:42,759 --> 00:51:45,461 SHORTER IN THE MUTANT HTT CELLS 976 00:51:45,461 --> 00:51:47,497 AND THAT'S SHOWED IN THE WESTERN 977 00:51:47,497 --> 00:51:49,299 BLOT AND ALSO ON THE QUANTIFIED 978 00:51:49,299 --> 00:51:53,837 AND THEN WE LOOK AT THE--THE--IN 979 00:51:53,837 --> 00:51:56,272 THE ANIMAL MODEL, IN THE FAR 980 00:51:56,272 --> 00:51:58,908 RIGHT, THIS IS--THIS IS WESTERN 981 00:51:58,908 --> 00:52:02,245 BLOT SHOWING THE WILD-TYPE AND 982 00:52:02,245 --> 00:52:04,180 ACTUALLY HUNTING TON MOUSE 983 00:52:04,180 --> 00:52:09,986 MODEL, AND THEN WE MAJORED MLH1 984 00:52:09,986 --> 00:52:12,055 EXPRESSION AND YOU CAN SEE IN 985 00:52:12,055 --> 00:52:16,626 THE WILD-TYPE IT WAS MUCH HIGHER 986 00:52:16,626 --> 00:52:20,430 COMPARED WITH THE HD-HUNTING TON 987 00:52:20,430 --> 00:52:21,097 PATIENT ANIMAL MODEL. 988 00:52:21,097 --> 00:52:25,401 SO THIS SHOWS THE SAME RESULT IN 989 00:52:25,401 --> 00:52:29,138 CELL LINES SO WE BELIEVE THAT 990 00:52:29,138 --> 00:52:39,682 INDEED THERE ARE MUTANT WITH THE 991 00:52:40,950 --> 00:52:43,887 PROTEIN, PROBABLY, NOW THIS MLH1 992 00:52:43,887 --> 00:52:47,757 DEGRADED THAT'S PROBABLY DO YOU 993 00:52:47,757 --> 00:52:49,125 TO PROTEIN-PROTEIN INTERACTION 994 00:52:49,125 --> 00:52:50,293 BECAUSE THE WILD-TYPE INTERACT 995 00:52:50,293 --> 00:52:52,362 WITH THE PROTEIN BUT THE MUTANT 996 00:52:52,362 --> 00:52:54,898 PROTEIN CANNOT WHICH NOW MAY BE 997 00:52:54,898 --> 00:52:56,532 A NOW MLH1 TO BE DEGREATED IN 998 00:52:56,532 --> 00:53:01,404 THE CELL. 999 00:53:01,404 --> 00:53:04,440 NEXT UP, PLEASE. 1000 00:53:04,440 --> 00:53:10,546 OKAY, YEAH, WE SHOW THAT THE DNA 1001 00:53:10,546 --> 00:53:13,316 GENERATION AND IN THIS 1002 00:53:13,316 --> 00:53:14,183 HTT MUTANT CELLS EMPLOY NEXT 1003 00:53:14,183 --> 00:53:16,452 SLIDE, PLEASE AND THEN THE 1004 00:53:16,452 --> 00:53:18,855 ACTIVATION OF CGAS, POSSIBLY AS 1005 00:53:18,855 --> 00:53:21,257 WE SHOWED IN THE MLH1 DEFICIENT 1006 00:53:21,257 --> 00:53:25,094 TUMOR CELLS AND YOU CAN SEE 1007 00:53:25,094 --> 00:53:28,798 HERE, IT'S--IT'S REALLY 1008 00:53:28,798 --> 00:53:30,767 ACTIVATION OF WHEN YOU MARK IT 1009 00:53:30,767 --> 00:53:35,004 ON AGONIST 1, OF COURSE, YOU 1010 00:53:35,004 --> 00:53:36,205 ARE--YOU ARE SITTING 1011 00:53:36,205 --> 00:53:39,242 PHOSPHORYLATION HAS DISAPPEARED 1012 00:53:39,242 --> 00:53:45,348 AND THIS IS IN BOTH--ON 1013 00:53:45,348 --> 00:53:51,254 THE--YEAH, WE TRIED TO SHOW, I 1014 00:53:51,254 --> 00:53:56,426 MEAN THERE ACTIVATION OF CGAS 1015 00:53:56,426 --> 00:53:58,695 PATHWAY IN THE MUTANT CELLS AND 1016 00:53:58,695 --> 00:54:00,463 THIS ALSO CAUSE THE CELL DIES, 1017 00:54:00,463 --> 00:54:03,266 WHERE ON THE RIGHT WE SHOW THE 1018 00:54:03,266 --> 00:54:07,136 CASP A SE 3 AND 9, AND ACTUALLY 1019 00:54:07,136 --> 00:54:09,672 CLEAVED IN THE MUTANT CELLS BUT 1020 00:54:09,672 --> 00:54:11,874 WHEN YOU'RE KNOCKING OUT AGONIST 1021 00:54:11,874 --> 00:54:14,110 1, THIS IS NOW HAPPENING 1022 00:54:14,110 --> 00:54:16,312 SUGGESTING JUST ALIKE A MISMATCH 1023 00:54:16,312 --> 00:54:19,682 REPAIR DEFICIENT CANCER CELLS, 1024 00:54:19,682 --> 00:54:24,253 THAT WOULD REQUIRE EXO1 TO 1025 00:54:24,253 --> 00:54:28,024 ACTIVATE THE CYTOSOLIC DNA AND 1026 00:54:28,024 --> 00:54:29,659 ALSO CGAS [INDISCERNIBLE]. 1027 00:54:29,659 --> 00:54:30,560 NEXT SLIDE, PLEASE. 1028 00:54:30,560 --> 00:54:36,132 SO HERE'S THE MODEL THAT I--WE 1029 00:54:36,132 --> 00:54:38,101 PUTTING TOGETHER AND ON THE LEFT 1030 00:54:38,101 --> 00:54:39,936 AND WITH THE NORMAL SITUATION, 1031 00:54:39,936 --> 00:54:43,639 AND AS I SHOWED EARLIER IS A 1032 00:54:43,639 --> 00:54:46,843 CONTROL OF THE EXO1'S TROLL WELL 1033 00:54:46,843 --> 00:54:55,952 BUT ON THE RIGHT, YOU CAN SEE 1034 00:54:55,952 --> 00:54:59,155 THE--THERE'S 2 PROBLEMS, 1 THE 1035 00:54:59,155 --> 00:55:00,089 HTT PROTEIN IS [INDISCERNIBLE] 1036 00:55:00,089 --> 00:55:02,558 TO THE MUTANT PROTEIN, THE 1037 00:55:02,558 --> 00:55:04,360 MUTANT PROTEIN DID NOT INTERACT 1038 00:55:04,360 --> 00:55:07,263 WITH EXO1 AND AT THE SAME TIME 1039 00:55:07,263 --> 00:55:12,135 MUTANT L PROTEINS DEGRADED SO 1040 00:55:12,135 --> 00:55:13,770 EXO1, SO MUTANT CONTROLS SO 1041 00:55:13,770 --> 00:55:17,140 THAT'S WHY CGAS IS IMPOSSIBLY 1042 00:55:17,140 --> 00:55:18,274 ACTIVATED. 1043 00:55:18,274 --> 00:55:22,478 IN NEURON CELLS IT CAUSE, I MEAN 1044 00:55:22,478 --> 00:55:25,314 OR ACTUALLY WITH APOP TOAZIS, SO 1045 00:55:25,314 --> 00:55:27,316 THAT'S WHY YOU CAUSING THE 1046 00:55:27,316 --> 00:55:30,153 PATHOLOGY TO THE PATIENT, NEXT 1047 00:55:30,153 --> 00:55:31,354 SLIDE, PLEASE. 1048 00:55:31,354 --> 00:55:32,522 WELL I'M ACTUALLY--IN THE NEXT I 1049 00:55:32,522 --> 00:55:39,595 WILL TRY TO GO FAST, SO IN THE 1050 00:55:39,595 --> 00:55:42,832 LAST MAYBE 2 MINUTES, I WILL TRY 1051 00:55:42,832 --> 00:55:45,435 TO TALK ABOUT ANOTHER ACTIVITY, 1052 00:55:45,435 --> 00:55:47,703 WE IDENTIFIED AND WE HAVE NOT 1053 00:55:47,703 --> 00:55:52,675 PUBLISHED YET WHICH ISY 1054 00:55:52,675 --> 00:55:57,814 SHORTENING THE MUTANT L, 1055 00:55:57,814 --> 00:55:59,315 FOUNDATION IN INFERTILITY. 1056 00:55:59,315 --> 00:56:03,186 NEXT SLIDE, PLEASE. 1057 00:56:03,186 --> 00:56:06,155 SO IT HAS BEEN SHARED ALMOST 30 1058 00:56:06,155 --> 00:56:09,192 YEARS AGO THAT MALE MICE 1059 00:56:09,192 --> 00:56:12,495 DEFECTIVE MLH1 OR PMS2 ARE 1060 00:56:12,495 --> 00:56:12,762 INFERTILE. 1061 00:56:12,762 --> 00:56:17,266 AND BECAUSE THESE ANIMALS CANNOT 1062 00:56:17,266 --> 00:56:19,202 HAVE A NORMAL MEIOSIS AND THEY 1063 00:56:19,202 --> 00:56:23,005 CANNOT HAVE A NORMAL SPERMAT O 1064 00:56:23,005 --> 00:56:25,308 GENESIS, AND THEY ALSO HAVE 1065 00:56:25,308 --> 00:56:28,878 SMALLER TESTIS, AND NOBODY KNOWS 1066 00:56:28,878 --> 00:56:31,514 WHAT REALLY--I MEAN, WHY THIS IS 1067 00:56:31,514 --> 00:56:31,747 CAUSED. 1068 00:56:31,747 --> 00:56:34,617 AND THEN AT A CERTAIN TIME, WE 1069 00:56:34,617 --> 00:56:40,156 NOTICE THAT THERE'S ANOTHER 1070 00:56:40,156 --> 00:56:42,225 ANIMAL THAT'S DEFECTIVE IN YAPP, 1071 00:56:42,225 --> 00:56:45,294 ARE ALSO SHOWING SOME MORE 1072 00:56:45,294 --> 00:56:47,797 TESTIS AND KIND THAT GENERATE 1073 00:56:47,797 --> 00:56:52,568 MATURE SPERM AND SO, IT'S ALSO 1074 00:56:52,568 --> 00:56:53,202 COMPLETELY INFERTILE. 1075 00:56:53,202 --> 00:56:57,507 SO NEXT SLIDE, PLEASE. 1076 00:56:57,507 --> 00:57:00,176 SO WE BELIEVE THAT THIS MAY BE 1077 00:57:00,176 --> 00:57:02,478 DOING SOMETHING, WELL IT MAY BE 1078 00:57:02,478 --> 00:57:03,913 DOING SOMETHING WITH YAPP, SO 1079 00:57:03,913 --> 00:57:08,684 FIRST ORDER IS THE YAPP, IT'S 1080 00:57:08,684 --> 00:57:10,520 ACTUALLY THE TRANSCRIPTION CO 1081 00:57:10,520 --> 00:57:14,724 ACTIVATOR AND IT'S A DOWN STREAM 1082 00:57:14,724 --> 00:57:16,893 EFFECT OF HIPPO PATHWAY WHICH 1083 00:57:16,893 --> 00:57:18,494 CONTROLS ORGAN GROWTH AND THE 1084 00:57:18,494 --> 00:57:18,761 QUEPMENT. 1085 00:57:18,761 --> 00:57:22,532 AND THIS IS WHAT A YAPP DOES. 1086 00:57:22,532 --> 00:57:26,235 SO YAPP NORMALLY CAN BE 1087 00:57:26,235 --> 00:57:28,871 PHOSPHORYLATED BY MASTER 1 OR 2, 1088 00:57:28,871 --> 00:57:36,746 AND THIS WILL BE DEGRADED IN 1089 00:57:36,746 --> 00:57:38,281 CYTOSOL, BUT OUR WORK GOES INTO 1090 00:57:38,281 --> 00:57:41,551 NUCLEUS AND THEN PLAY A CO 1091 00:57:41,551 --> 00:57:43,052 ACTIVELY, TRANSCRIBING CO ACTIVE 1092 00:57:43,052 --> 00:57:46,689 FUNCTION AND EXPRESS A NUMBER OF 1093 00:57:46,689 --> 00:57:49,559 YEENS INVOLVED IN ORGAN 1094 00:57:49,559 --> 00:57:50,860 DEVELOPMENT OR GROWTH EMPLOY SO 1095 00:57:50,860 --> 00:57:51,661 NEXT SLIDE, PLEASE. 1096 00:57:51,661 --> 00:57:55,097 SO WE BELIEVE THAT MLH1 PROBABLY 1097 00:57:55,097 --> 00:57:57,767 CONTROLS TESTIS DEVELOPMENT BY 1098 00:57:57,767 --> 00:58:00,670 REGULATING PATHWAY, SO THIS 1099 00:58:00,670 --> 00:58:03,539 OTHER WORK TEND TO POST 1100 00:58:03,539 --> 00:58:04,874 [INDISCERNIBLE] AND THIS CATANOT 1101 00:58:04,874 --> 00:58:05,341 PUBLISHED YET. 1102 00:58:05,341 --> 00:58:09,645 NEXT SLIDE, I WILL GO REAL 1103 00:58:09,645 --> 00:58:10,513 QUICK. 1104 00:58:10,513 --> 00:58:13,583 OKAY, THE FIRST IMMEDIATE 1105 00:58:13,583 --> 00:58:14,550 INTERACTION BETWEEN MUTANT 1106 00:58:14,550 --> 00:58:16,419 L-PROTEIN AND THE YAPP, AS YOU 1107 00:58:16,419 --> 00:58:19,622 CAN SEE HERE THIS IS A POA ASSAY 1108 00:58:19,622 --> 00:58:23,292 AND YOU CAN SEE WHEN THEY HAVE 1109 00:58:23,292 --> 00:58:24,927 THE MLH1 ON WITH THE YAPP AND 1110 00:58:24,927 --> 00:58:31,968 YOU CAN SEE THEY ARE LINKED 1111 00:58:31,968 --> 00:58:32,235 TOGETHER. 1112 00:58:32,235 --> 00:58:33,569 AND THE IMMUNO PRECIPITATION 1113 00:58:33,569 --> 00:58:35,671 ALSO SHOWED ON THE RIGHT THAT 1114 00:58:35,671 --> 00:58:38,874 THOSE YAPP OR IRPT ACTION WITH 1115 00:58:38,874 --> 00:58:41,744 THE MLH1, OVER YAPP, I MEAN 1116 00:58:41,744 --> 00:58:44,780 MOUSE 1 IS THE KINASE FOR YAPP, 1117 00:58:44,780 --> 00:58:47,483 THOSE 2 PROTEINS TOGETHER BUT IF 1118 00:58:47,483 --> 00:58:53,155 YOU USE MLH1, IT'S A BASE TO 1119 00:58:53,155 --> 00:58:54,423 IMMUNO PRECIPITATION, AND YOU 1120 00:58:54,423 --> 00:58:56,692 OWN IT FOR YAB AND THAT'S' THE 1121 00:58:56,692 --> 00:59:00,696 1, SO THIS SHOWS THIS 1122 00:59:00,696 --> 00:59:02,431 INTERACTION BETWEEN MLH1 AND THE 1123 00:59:02,431 --> 00:59:06,202 YAPP. 1124 00:59:06,202 --> 00:59:06,769 NEXT SLIDE, PLEASE. 1125 00:59:06,769 --> 00:59:08,571 SO WE KNOW HOW THIS INTERACTION 1126 00:59:08,571 --> 00:59:11,340 WORKS AND WE HOE THIS 1127 00:59:11,340 --> 00:59:15,278 INTERACTION ACTUALLY INHIBITS 1128 00:59:15,278 --> 00:59:16,712 YAPP POSFORALATION FOR LATS1. 1129 00:59:16,712 --> 00:59:20,316 YOU CAN SEE IN THE LAB, IN THE 1130 00:59:20,316 --> 00:59:22,018 WILD-TYPE, THE YAPP IS VERY 1131 00:59:22,018 --> 00:59:26,489 LITTLE, WHEN YOU KNOCK OUT MLH1, 1132 00:59:26,489 --> 00:59:29,659 PHOSPHORYLATION YOU SEE A LOT OF 1133 00:59:29,659 --> 00:59:34,330 PHOSPHORYLATED YAPP, SO 1134 00:59:34,330 --> 00:59:37,867 SUGGESTING THE MLH1 YAPP CAN 1135 00:59:37,867 --> 00:59:40,403 KEEP YAPP UNPHOSPHORYLATED AND 1136 00:59:40,403 --> 00:59:43,205 MAYBE YAPP HAS THE CHANCE TO 1137 00:59:43,205 --> 00:59:45,074 BUILD A TRANSCRIPTION PHASE SO 1138 00:59:45,074 --> 00:59:46,709 THAT IT'S AFFECTUATED, AND THEN 1139 00:59:46,709 --> 00:59:49,478 THEY ARE TESTED TO HOW THIS 1140 00:59:49,478 --> 00:59:51,447 PROTEIN INTERACTOR WORKS IN THE 1141 00:59:51,447 --> 00:59:52,715 INTERACTION DOMAIN, AND WE FIND 1142 00:59:52,715 --> 00:59:56,152 OR IN THE RIGHT HERE, JUST HERE 1143 00:59:56,152 --> 01:00:03,926 AND THEN WE SHOWED ON THE 1144 01:00:03,926 --> 01:00:06,495 BOTTOM, MLH1 WITH YAPP IN THE 1145 01:00:06,495 --> 01:00:09,065 WW1 AND WW2 DOMAIN, MOST DOMAIN, 1146 01:00:09,065 --> 01:00:13,402 IT'S ACTUALLY THE SAME DOMAIN 1147 01:00:13,402 --> 01:00:15,504 THAT YAPP INTERACT WITH LATS 1, 1148 01:00:15,504 --> 01:00:16,806 SO IT'S THE KINASE AND WE THINK 1149 01:00:16,806 --> 01:00:20,509 ABOUT THAT AND THIS INTERACTION 1150 01:00:20,509 --> 01:00:21,744 BLOCKS YAPP, [INDISCERNIBLE] 1151 01:00:21,744 --> 01:00:23,412 BECAUSE IT'S COMPETING FOR 1152 01:00:23,412 --> 01:00:25,781 BONDING. 1153 01:00:25,781 --> 01:00:26,449 NEXT SLIDE, PLEASE. 1154 01:00:26,449 --> 01:00:31,387 SO WE ALSO KNOW THAT YAPP, WE 1155 01:00:31,387 --> 01:00:35,191 CHECK, YAPP HAS SO FAR AS 1156 01:00:35,191 --> 01:00:38,728 NO--NOT BEEN IDENTIFIED WITH THE 1157 01:00:38,728 --> 01:00:41,197 NUCLEAR TRANSLOCATION SEQUENCE, 1158 01:00:41,197 --> 01:00:45,768 SO THE INTERACTION PROBABLY HELP 1159 01:00:45,768 --> 01:00:47,903 YAB, LOCATED 2 NUCLEACE AND THAT 1160 01:00:47,903 --> 01:00:50,673 YOU CAN SEE HERE WHEN YOU ARE IN 1161 01:00:50,673 --> 01:00:53,242 THE WILD-TYPE YOU WILL SAY YAPP, 1162 01:00:53,242 --> 01:00:55,845 OR YAPP WAS RED, YAPP, LOCATED 1163 01:00:55,845 --> 01:00:57,580 IN THE NUCLEUS, VERY WELL, SO 1164 01:00:57,580 --> 01:01:02,551 YOU KNOCK OUT MLH1, AND MOST OF 1165 01:01:02,551 --> 01:01:06,355 YAPP IS OUTSIDE OF THE NUCLEUS, 1166 01:01:06,355 --> 01:01:07,556 RESILIENCE STORE MLH1, YOU GET 1167 01:01:07,556 --> 01:01:09,992 THE NUCLEUS BACK IN AND 1168 01:01:09,992 --> 01:01:13,295 SUGGIESTING NLH1 DOES HELP YAPP 1169 01:01:13,295 --> 01:01:14,096 TRANSLOCATE INTO A NUCLEUS. 1170 01:01:14,096 --> 01:01:15,831 ON THE RIGHT, IF YOU ARE 1171 01:01:15,831 --> 01:01:19,902 KNOCKING OUT THE TRANSLOCATION 1172 01:01:19,902 --> 01:01:23,172 SEQUENCE IN MLH1 AND YAPP CANNOT 1173 01:01:23,172 --> 01:01:25,574 GO THROUGH THE NUCLEACE, BUT NOT 1174 01:01:25,574 --> 01:01:27,176 HUNDRED PERCENT, ANY TO 1175 01:01:27,176 --> 01:01:29,378 NUCLEACE, THEY SHOW NOW, THAT 1176 01:01:29,378 --> 01:01:30,379 IT'S NEIGHBORED SO YOU CAN SEE 1177 01:01:30,379 --> 01:01:37,987 THE BOTTOM 1 WHEN YOU KNOCK 1178 01:01:37,987 --> 01:01:39,588 OUT--YOU DISRUPT THE NUCLEAR 1179 01:01:39,588 --> 01:01:42,124 SEQUENCE AND YOU SEE THAT YAPP 1180 01:01:42,124 --> 01:01:44,994 IS OUTSIDE SO SUGGESTING THAT 1181 01:01:44,994 --> 01:01:47,096 MLH1 HELP YAPP ANY INTO 1182 01:01:47,096 --> 01:01:49,231 NUCLEACE. 1183 01:01:49,231 --> 01:01:49,832 NEXT SLIDE, PLEASE. 1184 01:01:49,832 --> 01:01:50,900 SO NOW WHAT WE WANT TO KNOW 1185 01:01:50,900 --> 01:02:01,377 WHETHER OR NOT THAT'S REALLY 1186 01:02:02,178 --> 01:02:03,412 HELP YAPP CONTROLLED GENE IN THE 1187 01:02:03,412 --> 01:02:05,981 TOP LEFT, YOU CAN SEE, WHEN YOU 1188 01:02:05,981 --> 01:02:08,250 KNOCK OUT MLH1, YOU DO NOT 1189 01:02:08,250 --> 01:02:09,251 AFFECT THE YAPP EXPRESSION, AND 1190 01:02:09,251 --> 01:02:12,021 THEY ARE THE SAME, BUT IF YOU 1191 01:02:12,021 --> 01:02:13,889 LOOK AT A YEEN THAT CONTROLLED 1192 01:02:13,889 --> 01:02:18,794 IT BY YAPP, FOR TRANSCRIPTION, 1193 01:02:18,794 --> 01:02:23,098 THE MLH1 KNOCK OUT DRAMATICALLY 1194 01:02:23,098 --> 01:02:24,033 REDUCE THE EXPRESSION, SO THOSE 1195 01:02:24,033 --> 01:02:26,569 ARE THE 1S YOU KNOW IT'S 1196 01:02:26,569 --> 01:02:28,604 DEVELOPMENT SO THAT'S PROBABLY 1197 01:02:28,604 --> 01:02:32,741 THE REASON WHY MLH1 DEFICIENCY 1198 01:02:32,741 --> 01:02:34,677 CANNOT GENERATE MATURE SPERM OR 1199 01:02:34,677 --> 01:02:38,714 THEN DEVELOPMENT OF TESTIS. 1200 01:02:38,714 --> 01:02:42,418 AND NOW, THIS MLH1 DEFICIENT 1201 01:02:42,418 --> 01:02:45,888 CELLS TO MICE, GO INTO IN PHOTO, 1202 01:02:45,888 --> 01:02:47,556 NEXT SLIDE, PLEASE. 1203 01:02:47,556 --> 01:02:50,025 THIS IS THE SUMMARY SLIDE FOR 1204 01:02:50,025 --> 01:02:50,860 THIS 1. 1205 01:02:50,860 --> 01:02:53,729 AND IN TERMAL CIRCUMSTANCES, IT 1206 01:02:53,729 --> 01:02:58,300 SHOWED IN THE LEFT, YOU HAVE 1207 01:02:58,300 --> 01:03:01,704 MLH1 INTACT WITH YAPP, WHICH 1208 01:03:01,704 --> 01:03:04,573 COMPETE WITH THE 1 FOR BONDING 1209 01:03:04,573 --> 01:03:08,611 WITH YAPP, AND THEN PROTECT YAPP 1210 01:03:08,611 --> 01:03:12,982 NOT FROM PHOSPHORYLATION SO YAPP 1211 01:03:12,982 --> 01:03:14,617 CAN GO INTO NUCLEUS SO THEY 1212 01:03:14,617 --> 01:03:16,151 CHANGE SOME OF THE DEVELOPMENT 1213 01:03:16,151 --> 01:03:17,786 GENE CAN EXPRESS, SO NOW TESTIS 1214 01:03:17,786 --> 01:03:20,956 TO HAVE A NORMAL DEVELOPMENT BUT 1215 01:03:20,956 --> 01:03:25,394 IF YOU DON'T HAVE THE MLH1 AND 1216 01:03:25,394 --> 01:03:28,297 YOU ARE YAPP OR IT'S MOSTLY 1217 01:03:28,297 --> 01:03:29,498 DEGRADED AND THEN ONLY A FEW OF 1218 01:03:29,498 --> 01:03:32,468 THEM CAN GO TO A NUCLEACE, SO 1219 01:03:32,468 --> 01:03:35,604 THAT'S WHY YOU ARE TESTING, IT'S 1220 01:03:35,604 --> 01:03:39,241 NOT NORMAL--NEXT SLIDE, PLEASE. 1221 01:03:39,241 --> 01:03:40,809 SO THIS CONVERT MY TALK AND IN 1222 01:03:40,809 --> 01:03:43,245 THE FINAL THERE, I WOULD LIKE TO 1223 01:03:43,245 --> 01:03:45,114 THANK THE PEOPLE WHO DID THE 1224 01:03:45,114 --> 01:03:47,149 WORK, I WOULD MENTION SOME OF 1225 01:03:47,149 --> 01:03:57,059 THEM, AND THE HIGHLIGHTED IN THE 1226 01:03:57,059 --> 01:03:58,594 SLIDE DID THE MAIORITY OF THE 1227 01:03:58,594 --> 01:04:01,597 WORK AND ALSO LISTS ALL THE 1228 01:04:01,597 --> 01:04:04,366 COLLABORATORS, MANY, MANY 1229 01:04:04,366 --> 01:04:08,504 RECRUITING, MANY COLLABORATORS 1230 01:04:08,504 --> 01:04:11,006 AND SO MANY OTHERS. 1231 01:04:11,006 --> 01:04:18,847 THANK YOU SO MUCH FOR LISTENING. 1232 01:04:18,847 --> 01:04:19,415 YEAH. 1233 01:04:19,415 --> 01:04:19,782 >> GREAT. 1234 01:04:19,782 --> 01:04:24,320 THANK YOU DR. LI FOR THE 1235 01:04:24,320 --> 01:04:24,787 WONDERFUL TALK. 1236 01:04:24,787 --> 01:04:27,690 WE WILL TART THE Q&A SECTION. 1237 01:04:27,690 --> 01:04:35,030 LET'S SEE, THERE IS A--KAREN DO 1238 01:04:35,030 --> 01:04:37,466 YOU WANT TO START? 1239 01:04:37,466 --> 01:04:37,933 >> YES. 1240 01:04:37,933 --> 01:04:42,204 SO PHILLIP JORDAN'S ASKED GOING 1241 01:04:42,204 --> 01:04:43,405 BACK TO THE CGAS PART OF THE 1242 01:04:43,405 --> 01:04:49,645 TALK THAT BASED ON THE CGAS 1243 01:04:49,645 --> 01:04:52,081 EXPRESSION,--SORRY EXPRESSION, 1244 01:04:52,081 --> 01:04:53,349 FOR IMMUNOTHERAPY, ARE THERE 1245 01:04:53,349 --> 01:04:54,883 ATTEMPTS TO INCREASE THESE 1246 01:04:54,883 --> 01:04:58,187 PROTEINS EXPERIMENTALLY TO SHOW 1247 01:04:58,187 --> 01:04:59,855 IT CAN POSITIVELY INFLUENCE 1248 01:04:59,855 --> 01:05:04,059 THEIR RESPONSE? 1249 01:05:04,059 --> 01:05:07,763 >> WELL, YEAH, THE ANSWER IS 1250 01:05:07,763 --> 01:05:12,768 YES, WE HAVE LIKE H6 CELL, THE 1251 01:05:12,768 --> 01:05:17,840 HTT6 CELL LINE IT HAS WILD-TYPE 1252 01:05:17,840 --> 01:05:20,876 CGAS GENE BUT UNFORT MATILY THE 1253 01:05:20,876 --> 01:05:27,049 PROMOTER OF CGAS GENE IS 1254 01:05:27,049 --> 01:05:28,117 [INDISCERNIBLE] AND THE GENE IS 1255 01:05:28,117 --> 01:05:33,055 NOT EXPRESSED BUT WHEN WE PUT IN 1256 01:05:33,055 --> 01:05:34,857 THIS H6 CELL IN ANIMAL MODEL FOR 1257 01:05:34,857 --> 01:05:39,328 TRYING TO DO THE IMMUNOTHERAPY 1258 01:05:39,328 --> 01:05:40,596 IS A LOT OF WORK BUT APPROXIMATE 1259 01:05:40,596 --> 01:05:42,498 WE RESTORE THE EXPRESSION OF 1260 01:05:42,498 --> 01:05:45,934 CGAS AND THEN IMMEDIATELY THE 1261 01:05:45,934 --> 01:05:49,638 RESPONDED TO ANTIBODY AGAINST 1262 01:05:49,638 --> 01:05:50,806 THE PDL1, PD1, THERAPY COMING 1263 01:05:50,806 --> 01:05:53,375 BACK O THAT'S REALLY--THAT'S THE 1264 01:05:53,375 --> 01:05:56,578 ONLY DATA WE HAVE, BUT IN HUMAN, 1265 01:05:56,578 --> 01:05:59,181 IN HUMAN PATIENT AS I SHOWED YOU 1266 01:05:59,181 --> 01:06:04,319 IN 1 OF THE SLIDES, THOSE 1267 01:06:04,319 --> 01:06:05,587 PATIENTS EXPRESS CGAS OR STING 1268 01:06:05,587 --> 01:06:08,991 VERY WELL AND THEY RESPONDED TO 1269 01:06:08,991 --> 01:06:13,529 IMMUNO THERAPY ANDARY 1270 01:06:13,529 --> 01:06:14,163 IMMUNOTHERAPY VERY POSITIVE. 1271 01:06:14,163 --> 01:06:16,465 >> OKAY, I WILL GO WITH THE 1272 01:06:16,465 --> 01:06:18,567 SECOND QUESTION WHICH IS 1273 01:06:18,567 --> 01:06:21,837 ACTUALLY ALSO FROM PHILLIP 1274 01:06:21,837 --> 01:06:23,072 JORDAN. 1275 01:06:23,072 --> 01:06:26,842 THE QUESTION IS, MLH1S OR 3, 1276 01:06:26,842 --> 01:06:29,244 INVOLVE IN CROSS OVER FORMATION 1277 01:06:29,244 --> 01:06:31,880 BETWEEN HOMOLOGOUS CHROMOSOME 1278 01:06:31,880 --> 01:06:33,148 DURING [INDISCERNIBLE] GENESIS 1279 01:06:33,148 --> 01:06:37,052 WHICH MEANS A SPERM O GENESIS 1280 01:06:37,052 --> 01:06:38,821 AND O-GENESIS, MLH1 OR 3 1281 01:06:38,821 --> 01:06:40,789 MUTATION RESULT IN ALMOST A 1282 01:06:40,789 --> 01:06:43,726 COMPLETE LOSS OF CROSS OVER 1283 01:06:43,726 --> 01:06:47,262 EVENTS WITH ALL SPERMAT O 1284 01:06:47,262 --> 01:06:49,465 SIGHTS, THIS LED TO ANUE 1285 01:06:49,465 --> 01:06:50,799 EMPLOYEDY AND INFER TITTY. 1286 01:06:50,799 --> 01:06:54,937 IS THE YAPP SHOWING ANY SIMILAR 1287 01:06:54,937 --> 01:06:56,538 LOCALIZATION, FOR EXAMPLE AT 1288 01:06:56,538 --> 01:06:59,441 CROSS OVER SITES OR DIM 1289 01:06:59,441 --> 01:07:03,545 SARDEFECTS WHERE GAP IS MUTATED. 1290 01:07:03,545 --> 01:07:04,646 NWOW, THAT'S A GOOD QUESTION, 1291 01:07:04,646 --> 01:07:13,188 BECAUSE I DO HAVE AN ASSAY, YET, 1292 01:07:13,188 --> 01:07:16,859 BUT I THINK--WELL I CANNOT BUT 1293 01:07:16,859 --> 01:07:20,796 GUESS BECAUSE YAPP IS DOING 1294 01:07:20,796 --> 01:07:23,031 SOMETHING, MANY NORMALLY IN 1295 01:07:23,031 --> 01:07:24,967 TRANSCRIPTION, BUT MLH1 I 1296 01:07:24,967 --> 01:07:28,103 BELIEVE, PROBABLY HELP, HELP 1297 01:07:28,103 --> 01:07:32,574 YAPP TO TRANSLOCATE FROM 1298 01:07:32,574 --> 01:07:35,110 CYTOSOLTO NUCLEACE BUT WITHOUT 1299 01:07:35,110 --> 01:07:39,281 THE MLH1, YAPP PROBABLY HAS A 1300 01:07:39,281 --> 01:07:41,517 VERY LIMITED AMOUNT OF YAPP OR 1301 01:07:41,517 --> 01:07:46,288 BE ABLE TO TRANSLOCATE IT INTO 1302 01:07:46,288 --> 01:07:50,192 NUCLEISOTOPE, SO THAT'S GOING TO 1303 01:07:50,192 --> 01:07:52,394 CAUSE THE--PARTICULARLY THE 1304 01:07:52,394 --> 01:07:53,495 TESTIS DEVELOPMENT ISSUES. 1305 01:07:53,495 --> 01:08:02,971 BUT I'M NOT SO SURE,--THERE 1306 01:08:02,971 --> 01:08:06,108 WAS--I WAS WITHOUT MLH1, MLH3, I 1307 01:08:06,108 --> 01:08:11,013 HAVE NO IDEA WHETHER OR NOT YAPP 1308 01:08:11,013 --> 01:08:14,917 INTERACTION WITH MLH1 ABSOLUTELY 1309 01:08:14,917 --> 01:08:17,886 OR PMS, 2, MLH3, I REALLY DON'T 1310 01:08:17,886 --> 01:08:19,354 KNOW WE HAVE NOT GOTTEN TO THAT 1311 01:08:19,354 --> 01:08:22,658 POINT YET BUT I WILL SAY THE 1312 01:08:22,658 --> 01:08:24,326 LOCALIZATION OF YAPP WITHOUT 1313 01:08:24,326 --> 01:08:30,699 MLH1, MOST OF THEM WILL BE IN 1314 01:08:30,699 --> 01:08:32,501 THE CYTOSOL, SO THEY'RE PROBABLY 1315 01:08:32,501 --> 01:08:37,639 IN THE CYTOSOL. 1316 01:08:37,639 --> 01:08:40,209 >> OKAY, NEXT QUESTION FROM 1317 01:08:40,209 --> 01:08:41,543 GEN-LI, VERY NICE TALK AND THERE 1318 01:08:41,543 --> 01:08:43,512 ARE 2 QUESTIONS. 1319 01:08:43,512 --> 01:08:45,280 HAVE YOU SHOWN THAT 1320 01:08:45,280 --> 01:08:47,349 [INDISCERNIBLE] ALPHA AND 1321 01:08:47,349 --> 01:08:48,450 HTTMODDULATE THE ACTIVITY TO 1322 01:08:48,450 --> 01:08:49,384 INCREASE THE SECTION, 2 1323 01:08:49,384 --> 01:08:50,118 QUESTIONS, WHICH PROTEIN IS 1324 01:08:50,118 --> 01:08:54,089 RESPONSIBLE FOR THE DECKER NA IN 1325 01:08:54,089 --> 01:08:54,723 RECOGNITION? 1326 01:08:54,723 --> 01:08:55,958 IS ANY [INDISCERNIBLE] PROTEIN 1327 01:08:55,958 --> 01:08:59,461 INVOLVED IN THIS PROCESS? 1328 01:08:59,461 --> 01:09:03,065 AND I WILL ASK YOU THE SECOND 1329 01:09:03,065 --> 01:09:03,632 QUESTION LATER. 1330 01:09:03,632 --> 01:09:07,169 >> SAY IT AGAIN, I DID NOT HEAR 1331 01:09:07,169 --> 01:09:07,536 CLEARLY. 1332 01:09:07,536 --> 01:09:10,138 >> WHICH PROTEIN IS RESPONSIBLE 1333 01:09:10,138 --> 01:09:14,543 FOR THE DNA END RECOGNITION? 1334 01:09:14,543 --> 01:09:19,281 DURING THE PROCESS OF WHERE 1335 01:09:19,281 --> 01:09:23,919 MUTANT L ALPHA IS BLOCKING HTT? 1336 01:09:23,919 --> 01:09:27,890 >> OH, MUTANT L-ALPHA, I THINK 1337 01:09:27,890 --> 01:09:31,360 IN THIS CASE MUTANT L-ALPHA 1338 01:09:31,360 --> 01:09:34,696 INTACT WITH HTTTHEY BOTH CAN 1339 01:09:34,696 --> 01:09:35,731 CONCUR NUCLEACE ACTIVITY, THEY 1340 01:09:35,731 --> 01:09:38,767 ONLY NEED 1 BUT IN CASE OF 1341 01:09:38,767 --> 01:09:41,937 MUTATING THE HTT, BECAUSE YOU 1342 01:09:41,937 --> 01:09:44,606 YOU NOTICE THE HTT ALREADY AND 1343 01:09:44,606 --> 01:09:49,378 NOW IT'S HTT AND THE 1344 01:09:49,378 --> 01:09:50,779 AGG NORMALLY, WILD-TYPE WITH THE 1345 01:09:50,779 --> 01:09:53,949 MLH1 WHICH WE BELIEVE WAS 1346 01:09:53,949 --> 01:09:54,583 STABILIZE MLH1. 1347 01:09:54,583 --> 01:09:58,387 BUT THE RESULT OF WILD-TYPE 1348 01:09:58,387 --> 01:10:01,123 PROTEIN, MLH1, PARTICULARLY I DO 1349 01:10:01,123 --> 01:10:05,727 KNOW IN BRAND CELLS MAYBE 1350 01:10:05,727 --> 01:10:08,497 FRAGILE TO DEGRADE IT-- 1351 01:10:08,497 --> 01:10:10,832 >> [INDISCERNIBLE] PROTEIN 1352 01:10:10,832 --> 01:10:14,102 SYSTEM NOT INVOLVED? 1353 01:10:14,102 --> 01:10:15,103 [INDISCERNIBLE] PROTEINS. 1354 01:10:15,103 --> 01:10:15,837 >> [INDISCERNIBLE] PROTEINS WE 1355 01:10:15,837 --> 01:10:19,574 HAVE NOT TESTED BUT YES, IT 1356 01:10:19,574 --> 01:10:21,043 COULD INTERAT WITH THE L-PROTEIN 1357 01:10:21,043 --> 01:10:22,444 BUT THAT WILL REQUIRE A 1358 01:10:22,444 --> 01:10:24,379 MISMATCH, SO YOU HAVE TO HAVE A 1359 01:10:24,379 --> 01:10:26,915 DNA, SO THAT THOSE 2 PROTEIN 1360 01:10:26,915 --> 01:10:28,750 PROBABLY GOING TO IMPACT VERY 1361 01:10:28,750 --> 01:10:31,253 WELL WITH THE MISTACH REPAIR 1362 01:10:31,253 --> 01:10:33,221 REACTION IN THE ABSENCE OF 1363 01:10:33,221 --> 01:10:35,757 MISMATCH, I DO NOT KNOW HOW IT 1364 01:10:35,757 --> 01:10:39,828 PROTEIN WILL BE A CHANCE TO GET 1365 01:10:39,828 --> 01:10:40,595 IT TOGETHER. 1366 01:10:40,595 --> 01:10:43,065 >> OKAY, THE FOLLOW UP QUESTION 1367 01:10:43,065 --> 01:10:44,466 IS HTTISSUING INVOLVEMENT NEURON 1368 01:10:44,466 --> 01:10:44,833 CELLS SPECIFIC? 1369 01:10:44,833 --> 01:10:47,202 OR IS IT A GENERAL MECHANISM 1370 01:10:47,202 --> 01:10:54,576 THAT'S ACTIVE IN ALL CELL TYPES 1371 01:10:54,576 --> 01:10:56,778 >> YEAH, I AM NOT A 1372 01:10:56,778 --> 01:10:57,145 NEUROBIOLOGIST. 1373 01:10:57,145 --> 01:10:58,847 I CANNOT ANSWER THIS QUESTION, 1374 01:10:58,847 --> 01:11:01,650 BUT AT THE LEAST IN THE CELL, WE 1375 01:11:01,650 --> 01:11:02,818 ARE STUDYING ALTHOUGH WE ARE 1376 01:11:02,818 --> 01:11:06,922 TRYING TO USE THE ANIMAL MODEL, 1377 01:11:06,922 --> 01:11:08,890 I MEAN DERIVED FROM NEURON 1378 01:11:08,890 --> 01:11:11,526 CELLS, BUT IN OTHER CELLS, EVEN 1379 01:11:11,526 --> 01:11:16,531 WITHIN THE CELL, WE FIND, YOU 1380 01:11:16,531 --> 01:11:18,600 STILL HAVE HTT AND THOSE 1381 01:11:18,600 --> 01:11:22,237 REACTION, WHAT I TOLD YOU IS 1382 01:11:22,237 --> 01:11:25,240 ACTUALLY HAPPENING IN NONNEURON 1383 01:11:25,240 --> 01:11:25,807 CELL AS WELL. 1384 01:11:25,807 --> 01:11:27,676 >> MAY I ASK A QUESTION OF MY 1385 01:11:27,676 --> 01:11:30,445 OWN AT THIS POINT DO WE ARE 1386 01:11:30,445 --> 01:11:31,646 TIME? 1387 01:11:31,646 --> 01:11:32,681 >> YEAH. 1388 01:11:32,681 --> 01:11:36,551 >> SO, AS YOU KNOW MLH1 IS 1389 01:11:36,551 --> 01:11:40,522 REQUIRED FOR EXPANSION IN 1390 01:11:40,522 --> 01:11:42,090 HUNTING TON'S DEC, AND AUTOKNOWN 1391 01:11:42,090 --> 01:11:43,992 THAT EXPANSION CONTINUES IN 1392 01:11:43,992 --> 01:11:47,929 NEURONS THROUGHOUT THE LIFETIME 1393 01:11:47,929 --> 01:11:50,499 OF THAT NEURON SO OBVIOUSLY MLH1 1394 01:11:50,499 --> 01:11:54,002 REMAINS EVEN THOUGH MUTANTS 1395 01:11:54,002 --> 01:11:55,537 HUNTING TON'S IS THERE. 1396 01:11:55,537 --> 01:11:56,938 CAN YOU SPECULATE WHY THAT IS? 1397 01:11:56,938 --> 01:12:00,275 SO YOU MIGHT EXPECT THAT, WELL, 1398 01:12:00,275 --> 01:12:01,810 ARE YOU SUGGESTING THAT 1399 01:12:01,810 --> 01:12:03,879 INCREASED MUTANT HUNTING TON 1400 01:12:03,879 --> 01:12:05,580 WOULD EVENTUALLY LEAD TO 1401 01:12:05,580 --> 01:12:08,784 EXPANSION TOPPING IN THAT CELL 1402 01:12:08,784 --> 01:12:09,084 TYPE? 1403 01:12:09,084 --> 01:12:10,452 BECAUSE THAT DOESN'T SEEM TO BE 1404 01:12:10,452 --> 01:12:15,724 WHAT'S HAPPENING IN NYEAH, THE 1405 01:12:15,724 --> 01:12:23,565 EXPANSION OF HUNTING TON OR 1406 01:12:23,565 --> 01:12:26,301 EXPANSION GOING TO CAUSE--WELL, 1407 01:12:26,301 --> 01:12:29,971 I WILL SAY, MUTANT L-PROTEIN 1408 01:12:29,971 --> 01:12:34,543 WILL NOT BE PROTECTED WHEN YOU 1409 01:12:34,543 --> 01:12:35,811 HAVE 1 MUTATED HUNTING TON 1410 01:12:35,811 --> 01:12:39,281 PROTEIN BECAUSE WE KNOW THEY ARE 1411 01:12:39,281 --> 01:12:41,116 HTT AND MLH1 INTACT VERY WELL 1412 01:12:41,116 --> 01:12:43,585 ALTHOUGH WE DO NOT--ESTIMATION 1413 01:12:43,585 --> 01:12:45,954 THAT WE HAVEN'T TESTERRED 1414 01:12:45,954 --> 01:12:52,294 WHETHER OR NOT, IN BRAND CELLS, 1415 01:12:52,294 --> 01:12:56,865 THE MLH1 PROTEIN, INTACT WAYS 1416 01:12:56,865 --> 01:12:58,567 CLEARLY THROUGH MLH3, I BELIEVE 1417 01:12:58,567 --> 01:13:00,335 WILL BUT AT THE LEAST, I MEAN, 1418 01:13:00,335 --> 01:13:04,106 MAYBE SOME PART OF THE MLH1 1419 01:13:04,106 --> 01:13:06,308 PROTEIN IS STILL ENACT WITH 1420 01:13:06,308 --> 01:13:09,511 HTTBUT WHEN YOU HAVE A MUTATED 1421 01:13:09,511 --> 01:13:12,514 EDGE, THAT PART OF THE MUTANT 1422 01:13:12,514 --> 01:13:16,284 L-IS NOT IMPACTED WITH THE 1423 01:13:16,284 --> 01:13:18,987 DEGRADED THAT'S PROBABLY THE 1424 01:13:18,987 --> 01:13:26,428 REASON WE SEE IN OUR STUDY. 1425 01:13:26,428 --> 01:13:27,729 >> OKAY, SO SINCE TIME IS 1426 01:13:27,729 --> 01:13:28,864 RESTRAINED WE WILL GO TO THE 1427 01:13:28,864 --> 01:13:31,700 LAST QUESTION OF THE DAY SO THIS 1428 01:13:31,700 --> 01:13:32,467 IS FROM [INDISCERNIBLE]. 1429 01:13:32,467 --> 01:13:35,570 DID YOU LOOK AT THE ROW OF POLO 1430 01:13:35,570 --> 01:13:41,443 LIKE CAIN ACE IN ACTIVATING MLH1 1431 01:13:41,443 --> 01:13:43,078 IN MALES IN STUDIES SUCH AS 1432 01:13:43,078 --> 01:13:45,413 CANNED MEN AND WOMEN ACTIVATES 1433 01:13:45,413 --> 01:13:48,216 MLH1 WITH THE INTERACTION WITH 1434 01:13:48,216 --> 01:13:49,918 EXO1, EXO1 IN THIS CASE MAY JUST 1435 01:13:49,918 --> 01:13:53,655 BE MEDIATE BE THE INTERACTION OF 1436 01:13:53,655 --> 01:13:57,459 CDC5 WITH MLH1? 1437 01:13:57,459 --> 01:14:01,263 >> WOW, THAT'S A QUESTION, I 1438 01:14:01,263 --> 01:14:02,631 REALLY, REALLY DON'T HAVE ANSWER 1439 01:14:02,631 --> 01:14:03,131 FOR. 1440 01:14:03,131 --> 01:14:04,733 SO WHAT THE ENZYME HE WAS 1441 01:14:04,733 --> 01:14:10,972 TALKING ABOUT, I DID NOT 1442 01:14:10,972 --> 01:14:19,181 HEAR--THE COMPONENT, THE ENZYME 1443 01:14:19,181 --> 01:14:19,981 >> THE COLO-LIKE PRK,. 1444 01:14:19,981 --> 01:14:22,918 >> NO, NO, I HAVE NO IDEA. 1445 01:14:22,918 --> 01:14:24,886 >> I DON'T HAVE A LOT OF 1446 01:14:24,886 --> 01:14:25,420 INFORMATION. 1447 01:14:25,420 --> 01:14:29,124 NO I CANNOT ANSWER THOSE 1448 01:14:29,124 --> 01:14:29,958 QUESTIONS. 1449 01:14:29,958 --> 01:14:30,225 >> OKAY. 1450 01:14:30,225 --> 01:14:32,561 OKAY, SO, I THINK THAT'S PRETTY 1451 01:14:32,561 --> 01:14:34,763 MUCH ALL THE QUESTIONS. 1452 01:14:34,763 --> 01:14:36,798 SO, AGAIN, PLEASE JOIN ME TO 1453 01:14:36,798 --> 01:14:41,369 THANK DR. LI AGAIN FOR THIS 1454 01:14:41,369 --> 01:14:42,771 WONDERFUL TALK AND PLEASE BE 1455 01:14:42,771 --> 01:14:46,241 AWARE THAT DR. LI IS IN CHINESE 1456 01:14:46,241 --> 01:14:49,244 TIME ZONE SO IT'S ALREADY 1457 01:14:49,244 --> 01:14:49,978 MIDNIGHT, SO, AGAIN, PLEASE 1458 01:14:49,978 --> 01:14:51,613 THANK YOU FOR THE WONDERFUL TALK 1459 01:14:51,613 --> 01:14:55,317 AND TO HAVE A GREAT GOOD REST. 1460 01:14:55,317 --> 01:15:00,755 >> OKAY, THANK YOU SO MUCH, 1461 01:15:00,755 --> 01:15:01,523 KAREN AND CHEN. 1462 01:15:01,523 --> 01:15:02,224 THANK YOU. 1463 01:15:02,224 --> 01:15:03,325 >> THANK YOU NTHANK YOU. 1464 01:15:03,325 --> 01:15:06,561 >> THANK YOU TAKE CARE. 1465 01:15:06,561 --> 01:15:16,738 >> BYE.