1 00:00:05,794 --> 00:00:08,430 SO WELCOME, EVERYBODY 2 00:00:08,430 --> 00:00:10,265 TO TODAY'S WALS SEMINAR. 3 00:00:10,265 --> 00:00:12,868 FOR THOSE OF YOU ONLINE, I 4 00:00:12,868 --> 00:00:16,071 SHOULD MENTION OR FOR ANYONE 5 00:00:16,071 --> 00:00:17,606 WHO'S GETTING CME CREDITS FOR 6 00:00:17,606 --> 00:00:21,610 THIS, REMEMBER THAT THE CODE IS 7 00:00:21,610 --> 00:00:21,910 57681. 8 00:00:21,910 --> 00:00:24,212 SO THAT'S THE CODE YOU WILL HAVE 9 00:00:24,212 --> 00:00:27,649 TO ENTER TO GET CREDIT. 10 00:00:27,649 --> 00:00:29,484 FOR THOSE OF YOU WHO DON'T KNOW 11 00:00:29,484 --> 00:00:35,490 ME, I'M BEN WHITE, I'M A MEMBER 12 00:00:35,490 --> 00:00:36,391 OF THE DROSOPHILA NEUROSCIENCE 13 00:00:36,391 --> 00:00:38,293 INTEREST GROUP AT NIH AND OUR 14 00:00:38,293 --> 00:00:39,428 GROUP IS COSPONSORED TOGETHER 15 00:00:39,428 --> 00:00:41,196 WITH THE WOMEN'S SCIENTISTS 16 00:00:41,196 --> 00:00:43,632 ADVISORS COMMITTEE OF TODAY'S 17 00:00:43,632 --> 00:00:48,904 WALS SPEAKER DR. DANIELA 18 00:00:48,904 --> 00:00:51,006 ZARNESCU, OF THE COLLEGE OF PENN 19 00:00:51,006 --> 00:00:52,474 STATE MEDICINE. 20 00:00:52,474 --> 00:00:56,712 OT DROSOPHILA SIDE I SHOULD SAY 21 00:00:56,712 --> 00:00:58,847 THAT DR. ZARNESCU, WAS AN 22 00:00:58,847 --> 00:01:00,916 APPROPRIATE PICK FOR THIS 23 00:01:00,916 --> 00:01:03,719 SEMINAR SERIES AND HER WORK 24 00:01:03,719 --> 00:01:06,488 SHOWS THE POWER OF WORK OF 25 00:01:06,488 --> 00:01:09,725 STUDYING FLIES TO BRIDGE THE GAP 26 00:01:09,725 --> 00:01:11,093 IN UNDERLYING HUMAN PROCESSES 27 00:01:11,093 --> 00:01:12,361 AND HUMAN DISEASE. 28 00:01:12,361 --> 00:01:15,430 AND MOST OF US WHO WORK IN FLIES 29 00:01:15,430 --> 00:01:21,837 KNOW THAT OUR WORK ALTHOUGH IT'S 30 00:01:21,837 --> 00:01:24,806 AIMED AT BIOMEDICAL ADVANCES, 31 00:01:24,806 --> 00:01:28,543 OFTEN WE WORK PRETTY FAR FROM 32 00:01:28,543 --> 00:01:32,214 THE CLINIC AND YOU WILL SEE THAT 33 00:01:32,214 --> 00:01:32,848 DR. ZARNESCU'S RESEARCH IS 34 00:01:32,848 --> 00:01:35,217 DIFFERENT IN A SENSE THAT IT'S 35 00:01:35,217 --> 00:01:39,187 VERY MUCH DIRECTED AT 36 00:01:39,187 --> 00:01:40,389 UNDERSTANDING THE CLINICAL 37 00:01:40,389 --> 00:01:41,690 NEURAL PATHOLOGIES BASED ON WORK 38 00:01:41,690 --> 00:01:43,325 THAT DONE'S IN THE FLY. 39 00:01:43,325 --> 00:01:45,994 SO HER LAB DEVELOPED FLY MODELS 40 00:01:45,994 --> 00:01:47,929 FOR ALS AND FRONTAL LOBE 41 00:01:47,929 --> 00:01:49,798 DEMENTIA AND THEN SHE USED THOSE 42 00:01:49,798 --> 00:01:52,734 MODELS TO GAIN INSIGHT INTOS 43 00:01:52,734 --> 00:01:53,568 MOLECULAR MECHANISMS THAT THEN 44 00:01:53,568 --> 00:01:57,572 COULD BE TESTED IN HUMAN TISSUES 45 00:01:57,572 --> 00:01:58,540 AND PATIENT DERIVED IPSC 46 00:01:58,540 --> 00:01:59,474 NEURONS. 47 00:01:59,474 --> 00:02:01,777 SO USING THAT STRATEGY HER LAB 48 00:02:01,777 --> 00:02:05,614 HAS DISCOVERED NOVEL PATHOGENIC 49 00:02:05,614 --> 00:02:06,882 ROLES FOR A VARIETY OF MUTANT 50 00:02:06,882 --> 00:02:08,617 PROEN TOOS WHICH I AM SURE YOU 51 00:02:08,617 --> 00:02:10,152 WILL HEAR PLENTY MORE ABOUT IN 52 00:02:10,152 --> 00:02:11,586 HER TALK TODAY. 53 00:02:11,586 --> 00:02:12,754 IT'S WORTH POINTING OUT HOWEVER, 54 00:02:12,754 --> 00:02:14,856 THAT IN ADDITION TO USING THE 55 00:02:14,856 --> 00:02:17,392 FLY TO ELUCIDATE THE KEY 56 00:02:17,392 --> 00:02:21,163 MOLECULES INVOLVED IN PROCESSES, 57 00:02:21,163 --> 00:02:22,831 UNDERLYING NEUROPATHOGENESIS 58 00:02:22,831 --> 00:02:24,566 THAT DR. ZARNESCU IS ALSO 59 00:02:24,566 --> 00:02:26,701 LEVERAGING THE FLY MODEL AS A 60 00:02:26,701 --> 00:02:28,670 FIRST PASS TOOL FOR DRUG 61 00:02:28,670 --> 00:02:30,939 DISCOVERY, KREENING FOR SMALL 62 00:02:30,939 --> 00:02:32,374 MOLECULES THAT MAY PROVIDE 63 00:02:32,374 --> 00:02:33,442 THERAPEUTIC BENEFITS AND HUMANS, 64 00:02:33,442 --> 00:02:38,580 SO JUST AS A BIT OF BACKGROUND 65 00:02:38,580 --> 00:02:40,048 DR. ZARNESCU STUDIED PHYSICS IN 66 00:02:40,048 --> 00:02:44,853 ROMANIA BEFORE COMING TO THE 67 00:02:44,853 --> 00:02:46,822 U.S. BEFORE GETTING HER DEGREE 68 00:02:46,822 --> 00:02:47,889 IN MOLECULAR BIOLOGY AT PENN 69 00:02:47,889 --> 00:02:48,089 STATE. 70 00:02:48,089 --> 00:02:50,992 SHE WENT ON TO DO POST DOCTORAL 71 00:02:50,992 --> 00:02:52,928 WORK AT EMORY WITH KEVIN MOSES 72 00:02:52,928 --> 00:02:54,596 WITH THE FRAGILE X PROTEIN OF 73 00:02:54,596 --> 00:02:56,498 DROSOPHILA AND THEN JOINED THE 74 00:02:56,498 --> 00:02:58,033 FACULTY AT UNIVERSITY OF ARIZONA 75 00:02:58,033 --> 00:02:59,234 IN 2006. 76 00:02:59,234 --> 00:03:00,268 AND IT'S INTERESTING AT LEAST I 77 00:03:00,268 --> 00:03:04,306 WAS INTERESTED IN LOOKING AT HER 78 00:03:04,306 --> 00:03:06,141 CEV, TO SEE THAT THE GROWING 79 00:03:06,141 --> 00:03:07,742 CLINICAL RELEVANCE OF HER WORK 80 00:03:07,742 --> 00:03:10,645 COULD ALMOST BE FOLLOWED IN HER 81 00:03:10,645 --> 00:03:11,580 TRAJECTORY FROM ASSISTANT 82 00:03:11,580 --> 00:03:13,415 PROFESSOR TO FULL PROFESSOR AT 83 00:03:13,415 --> 00:03:15,317 THE UNIVERSITY OF ARIZONA IN THE 84 00:03:15,317 --> 00:03:17,686 JOINT APPOINTMENTS THAT SHE ADO 85 00:03:17,686 --> 00:03:18,086 YOU MEANULATED. 86 00:03:18,086 --> 00:03:20,222 SO BY THE TIME SHE LEFT THE 87 00:03:20,222 --> 00:03:22,624 UNIVERSITY OF ARIZONA IN 2022, 88 00:03:22,624 --> 00:03:25,527 TO GO BACK TO PENN STATE, SHE 89 00:03:25,527 --> 00:03:27,262 HAD APPOINTMENTS NOT ONLY IN HER 90 00:03:27,262 --> 00:03:29,130 PRIMARY DEPARTMENT OF MOLECULAR 91 00:03:29,130 --> 00:03:31,199 AND CELLULAR BIOLOGY, BUT ALSO 92 00:03:31,199 --> 00:03:33,602 IN NEUROLOGY, CELLULAR AND 93 00:03:33,602 --> 00:03:35,370 MOLECULAR MEDICINE, NEUROSCIENCE 94 00:03:35,370 --> 00:03:35,704 AND GENETICS. 95 00:03:35,704 --> 00:03:38,306 SO I THINK THAT SPEAKS TO THE 96 00:03:38,306 --> 00:03:40,809 BROAD IMPACT OF HER WORK. 97 00:03:40,809 --> 00:03:47,148 SO FINALLY AS I MENTIONED, 98 00:03:47,148 --> 00:03:48,984 DR. ZARNESCU, IS A -- IS ALSO 99 00:03:48,984 --> 00:03:52,187 SPONSORED TODAY BY THE WOMEN 100 00:03:52,187 --> 00:03:53,355 SCIENTISTS ADVISOR'S COMMITTEE 101 00:03:53,355 --> 00:03:55,090 AND THAT REFLECTS HER LONG 102 00:03:55,090 --> 00:03:57,392 STANDING COMMITMENT TO MENTORING 103 00:03:57,392 --> 00:03:58,527 EDUCATION AND TRAINING, FOR 104 00:03:58,527 --> 00:04:01,296 WHICH SHE HAS WON NUMEROUS 105 00:04:01,296 --> 00:04:01,630 AWARDS. 106 00:04:01,630 --> 00:04:02,697 SHE'S CURRENTLY ASSOCIATE DEAN 107 00:04:02,697 --> 00:04:04,766 FOR GRADUATE EDUCATION AND POST 108 00:04:04,766 --> 00:04:06,134 DOCTORAL TRAINING AT PENN STATE 109 00:04:06,134 --> 00:04:07,836 COLLEGE OF MEDICINE AND I WANT 110 00:04:07,836 --> 00:04:09,871 TO MENTION THAT SHE KINDLY 111 00:04:09,871 --> 00:04:11,940 OFFERED TO ALSO LEAD A SPECIAL 112 00:04:11,940 --> 00:04:14,809 CAREER WORKSHOP TOMORROW FOR 113 00:04:14,809 --> 00:04:16,211 FELLOWS, THAT'S ENTITLED STAYING 114 00:04:16,211 --> 00:04:18,079 THE COURSE, LESSONS IN BUILDING 115 00:04:18,079 --> 00:04:18,780 RESILIENCE. 116 00:04:18,780 --> 00:04:20,515 SO FOR FELLOWS WHO WOULD LIKE TO 117 00:04:20,515 --> 00:04:21,650 ATTEND THE WORKSHOP, IT'S 118 00:04:21,650 --> 00:04:24,786 TOMORROW AT 2:00 P.M. IN 119 00:04:24,786 --> 00:04:25,654 BUILDING 40. 120 00:04:25,654 --> 00:04:27,923 SPOTLIGHT TODAY OF COURSE IS ON 121 00:04:27,923 --> 00:04:29,424 HER SCIENCE, HER EXCITING 122 00:04:29,424 --> 00:04:31,593 SCIENCE AND SO, WITHOUT FURTHER 123 00:04:31,593 --> 00:04:33,695 ADO, I WANT TO TURN THE PODIUM 124 00:04:33,695 --> 00:04:39,301 OVER TO DR. ZARNESCU, WHO'S TALK 125 00:04:39,301 --> 00:04:43,071 TITLE PERHAPS WITH APOLOGIES TO 126 00:04:43,071 --> 00:04:44,906 ROBERT BERNE AND JOHN STEIN BECK 127 00:04:44,906 --> 00:04:47,976 IS OF FLIES AND MEN-HARNESSING 128 00:04:47,976 --> 00:04:50,312 THE POWER OF DROSOPHILA TO 129 00:04:50,312 --> 00:04:51,813 DECIPHER THE MOLECULAR 130 00:04:51,813 --> 00:04:57,218 MECHANISMS OF NEURO 131 00:04:57,218 --> 00:05:00,555 DEGENERATEDDATION GENERATEDDATI- 132 00:05:00,555 --> 00:05:01,022 DEGENERATION. 133 00:05:01,022 --> 00:05:03,625 SO PLEASE HELP ME WELCOME HER. 134 00:05:03,625 --> 00:05:05,727 >> CAN YOU HEAR ME OKAY, IT 135 00:05:05,727 --> 00:05:06,995 FEELS SPECIAL TO BE HERE SO 136 00:05:06,995 --> 00:05:08,296 THANK YOU MUCH FOR HAVING ME, 137 00:05:08,296 --> 00:05:09,898 EVERYONE AT MY INSTITUTION ASKS 138 00:05:09,898 --> 00:05:11,666 ME ARE YOU SURE THE SEMINAR IS 139 00:05:11,666 --> 00:05:14,102 STILL ON, AND I SAID AS FAR AS I 140 00:05:14,102 --> 00:05:16,638 KNOW YES, SO, I WAS REALLY 141 00:05:16,638 --> 00:05:18,006 IMPRESSED TO SEE THIS, I CAN 142 00:05:18,006 --> 00:05:19,541 TAKE THESE BACK AND PROVE THAT I 143 00:05:19,541 --> 00:05:21,576 WAS HERE AND THE SEMINAR TOOK 144 00:05:21,576 --> 00:05:25,914 PLACE IN CASE THEY'RE WONDERING 145 00:05:25,914 --> 00:05:26,715 WHAT'S GOING ON. 146 00:05:26,715 --> 00:05:29,517 SO THANK YOU VERY MUCH AGAIN, I 147 00:05:29,517 --> 00:05:31,720 PUT TOGETHER PERHAPS TOO MANY 148 00:05:31,720 --> 00:05:32,921 SLIDES SO A MAY NOT GO THROUGH 149 00:05:32,921 --> 00:05:36,324 ALL OF THEM BUT I DO WANT TO SAY 150 00:05:36,324 --> 00:05:39,561 THAT I LOVE FLIES AND I WAS 151 00:05:39,561 --> 00:05:41,062 ATTRACTED BY THE POWER OF FLY 152 00:05:41,062 --> 00:05:42,597 GENETICS WHEN I STUDIED GRADUATE 153 00:05:42,597 --> 00:05:43,431 SCHOOL AND THEN I DISCOVERED 154 00:05:43,431 --> 00:05:45,066 WHEN I WAS A POST DOC THAT THE 155 00:05:45,066 --> 00:05:47,969 FLY CAN BE A WONDERFUL SYSTEM TO 156 00:05:47,969 --> 00:05:48,970 UNDERSTAND BASIC MECHANISMS OF 157 00:05:48,970 --> 00:05:51,272 DISEASE AND I WAS WORKING ON 158 00:05:51,272 --> 00:05:52,574 FRAGILE X SYNDROME AT THE TIME 159 00:05:52,574 --> 00:05:56,378 AND IN MY LAB WE MADE THE SWITCH 160 00:05:56,378 --> 00:05:57,679 TO AGE DEPENDENT 161 00:05:57,679 --> 00:05:58,246 NEURODEGENERATIVE DISEASES 162 00:05:58,246 --> 00:06:00,382 BECAUSE AS YOU CAN TELL AGING 163 00:06:00,382 --> 00:06:01,683 AND INEVITABLE AND THAT BECAME 164 00:06:01,683 --> 00:06:04,319 RELEVANT AND INTERESTING TO ME 165 00:06:04,319 --> 00:06:05,086 AT THE TIME. 166 00:06:05,086 --> 00:06:07,489 SO I'M GOING TO TRY TO DO A BIT 167 00:06:07,489 --> 00:06:08,356 OF A TOUR DEFORCE THROUGH SOME 168 00:06:08,356 --> 00:06:12,761 OF THE WORK WE'VE DONE SINCE WE 169 00:06:12,761 --> 00:06:15,163 DEVELOPED FLY MODELS OF LATERAL 170 00:06:15,163 --> 00:06:17,232 SCLEROSIS AND I WOULD LIKE TO 171 00:06:17,232 --> 00:06:19,768 SHARE NEW AND UNPUBLISHED DATA 172 00:06:19,768 --> 00:06:21,169 AND I LOOK FORWARD TO GETTING 173 00:06:21,169 --> 00:06:22,904 FEEDBACK FROM ALL OF YOU AND 174 00:06:22,904 --> 00:06:23,938 ALSO PLEASE FEEL FREE TO 175 00:06:23,938 --> 00:06:25,106 INTERRUPT AND ASK QUESTIONS AT 176 00:06:25,106 --> 00:06:27,542 ANY TIME AND THAT LIGHT IS A BIT 177 00:06:27,542 --> 00:06:32,947 BLINDING SO IF I SQUINT, IT'S 178 00:06:32,947 --> 00:06:34,683 BECAUSE OF THAT. 179 00:06:34,683 --> 00:06:36,384 BUT -- SO NEURODEGENERATIVE 180 00:06:36,384 --> 00:06:39,387 DISORDERS ARE A GROUP OF 181 00:06:39,387 --> 00:06:41,256 HETEROGENEOUS DISEASES AND I WAS 182 00:06:41,256 --> 00:06:44,559 REALLY GLAD TO SEE THIS REVIEW 183 00:06:44,559 --> 00:06:47,896 THAT REALLY PUTS TOGETHER AWL 184 00:06:47,896 --> 00:06:48,430 HARKERS NEURODEGENERATIVE 185 00:06:48,430 --> 00:06:49,297 DISEASES VERY MUCH LIKE THE 186 00:06:49,297 --> 00:06:50,365 CANCER FIELD HAS DONE A LONG 187 00:06:50,365 --> 00:06:52,534 TIME AGO AND I LIKE TO 188 00:06:52,534 --> 00:06:58,373 PERSPECTIVE BECAUSE IT'S REALLY 189 00:06:58,373 --> 00:07:00,942 HIGHLIGHTS THE FACT THAT 190 00:07:00,942 --> 00:07:01,676 NEURODEGENERATIVE DISORDERS 191 00:07:01,676 --> 00:07:04,879 SHARE ALTERED COMMON PATHWAYS 192 00:07:04,879 --> 00:07:06,881 THAT INCLUDE ABNORMALITIES 193 00:07:06,881 --> 00:07:10,318 BROIGATION OF PROTEINS, INSIGNE 194 00:07:10,318 --> 00:07:12,921 APTIC DYSFUNCTION, DEFECTING 195 00:07:12,921 --> 00:07:15,990 PROTEIN COMPLEXIO STASIS, 196 00:07:15,990 --> 00:07:17,058 CYTOSKELETAL ABNORMALITIES 197 00:07:17,058 --> 00:07:18,927 AFFECTING METABOLISM AND ENERGY 198 00:07:18,927 --> 00:07:20,795 HOMEOSTASIS, DEFECT IN DNA AND 199 00:07:20,795 --> 00:07:25,100 RNA, AS WELL AS INFLAMMATION 200 00:07:25,100 --> 00:07:26,868 THAT ULTIMATELY LEADS TO 201 00:07:26,868 --> 00:07:29,738 NEURONAL DETH -- DEAG. 202 00:07:29,738 --> 00:07:31,806 AND I THINK THIS SCHEME PROVIDES 203 00:07:31,806 --> 00:07:35,410 A SCHEME TO THINK MORE 204 00:07:35,410 --> 00:07:37,712 WHOLISTICALLY OF THE 205 00:07:37,712 --> 00:07:38,680 NEURODEGENERATIVE DISORDIERS 206 00:07:38,680 --> 00:07:40,882 LIKE ALS, OR PARKINSONS THAT I 207 00:07:40,882 --> 00:07:41,950 WORK ON, BECAUSE ULTIMATE GOAL 208 00:07:41,950 --> 00:07:43,017 MITTLY THEY SHARE ALL OF THIS 209 00:07:43,017 --> 00:07:44,285 AND IT'S BECOME MORE AND MORE 210 00:07:44,285 --> 00:07:46,020 CLEAR IN RECENT YEARS THAT AS WE 211 00:07:46,020 --> 00:07:49,991 UNDERSTAND THE PATHOMECHANISMS 212 00:07:49,991 --> 00:07:51,593 IN 1 DISEASE THEY BECOME 213 00:07:51,593 --> 00:07:52,660 APPLICABLE OR PROVIDE INSIGHT 214 00:07:52,660 --> 00:07:57,732 INTOS WHAT WE CALL RELATED 215 00:07:57,732 --> 00:08:00,401 NEURODEGENERATIVE DISORDERS. 216 00:08:00,401 --> 00:08:03,271 IS THAT A LASER POINTER OR 217 00:08:03,271 --> 00:08:04,906 SHOULD I -- I WILL USE THE 218 00:08:04,906 --> 00:08:05,273 CURSOR, OKAY. 219 00:08:05,273 --> 00:08:08,109 SO IN NIGH LAB VERMEN INFECTED 220 00:08:08,109 --> 00:08:10,945 BEEN FOCUSING ON MI LATERAL 221 00:08:10,945 --> 00:08:12,547 SCLEROSIS SINCE 2008. 222 00:08:12,547 --> 00:08:13,615 IT'S A PROGRESSIVE 223 00:08:13,615 --> 00:08:15,216 NEURODEGENERATIVE DISORDER WITH 224 00:08:15,216 --> 00:08:16,551 A PREVALENCE OF 43 OR HIGHER 225 00:08:16,551 --> 00:08:18,052 DEPENDING ON WHAT PAPER YOU 226 00:08:18,052 --> 00:08:19,654 READ, A IN A HUNDRED THOUSAND 227 00:08:19,654 --> 00:08:19,888 PEOPLE. 228 00:08:19,888 --> 00:08:22,991 AGE IS THE BIGGEST RISK FACTOR, 229 00:08:22,991 --> 00:08:24,659 AND I'VE HAD REALLY WONDERFUL 230 00:08:24,659 --> 00:08:25,960 TALKS IN MORNING ABOUT AGING AND 231 00:08:25,960 --> 00:08:29,798 WHAT DOES THAT REALLY MEAN. 232 00:08:29,798 --> 00:08:32,400 THE AVERAGE LIFE SPAN IS ABOUT 2 233 00:08:32,400 --> 00:08:34,502 AND HALF YEARS FOR PATIENTS POST 234 00:08:34,502 --> 00:08:37,138 DIAGNOSIS, AND THERE ARE ONLY 4 235 00:08:37,138 --> 00:08:40,308 FDA APPROVED DRUGS THAT HAVE 236 00:08:40,308 --> 00:08:41,476 VARIOUS DEGREES OF IMPACT ON THE 237 00:08:41,476 --> 00:08:44,312 PROGRESSION OF THE DISEASE. 238 00:08:44,312 --> 00:08:47,715 WHAT IS REALLY REMARKABLE ABOUT 239 00:08:47,715 --> 00:08:49,684 ALS AND BRINGS AGAIN THE HILIGHT 240 00:08:49,684 --> 00:08:51,886 BACK ON THESE HALLMARKS OF 241 00:08:51,886 --> 00:08:54,322 NEURODEGENERATION IS THIS RNA 242 00:08:54,322 --> 00:08:55,256 BYPASSING PROTEIN CALLED TDP43. 243 00:08:55,256 --> 00:08:57,826 WHICH WE THINK OF AS A COMMON 244 00:08:57,826 --> 00:09:01,830 DENOMINATOR IN ALS AS WELL AS 245 00:09:01,830 --> 00:09:03,131 RELATED NEURODEGENERATIVE 246 00:09:03,131 --> 00:09:03,398 DISORDERS. 247 00:09:03,398 --> 00:09:07,302 IT'S AN AUTONOMIC BINDING 248 00:09:07,302 --> 00:09:09,270 PROTEIN WHEN WITH 2 DIFFERENT 249 00:09:09,270 --> 00:09:13,374 ROM DOMAINS AND A C-TERMINUS 250 00:09:13,374 --> 00:09:13,708 STRUCTURE. 251 00:09:13,708 --> 00:09:16,377 AND I'M HIGHLIGHTING THE FACT 252 00:09:16,377 --> 00:09:19,547 THAT PATIENTS IN TDP43, THAT 253 00:09:19,547 --> 00:09:21,649 ONLY REPRESENT 2-4% OF THE ALS 254 00:09:21,649 --> 00:09:22,350 POPULATION, THIS MUTATION'S 255 00:09:22,350 --> 00:09:25,153 CLUOF THENER THE C-TERMINUS 256 00:09:25,153 --> 00:09:26,387 DOMAIN OR UNSTRUCTURED DOMAIN 257 00:09:26,387 --> 00:09:29,057 AND THERE'S EVIDENCE THAT THESE 258 00:09:29,057 --> 00:09:31,759 MUTATIONS MAKE TDP 43, WHICH IS 259 00:09:31,759 --> 00:09:33,795 IN ITS WILD-TYPE FORM PRONE TO 260 00:09:33,795 --> 00:09:34,729 ABNORMALITIES BROIGATION, EVEN 261 00:09:34,729 --> 00:09:36,097 MORE BROWN TO ABNORMALITIES 262 00:09:36,097 --> 00:09:37,966 BROIGATION, SO THESE MUTATIONS 263 00:09:37,966 --> 00:09:39,500 ACCELERATE THE FORMATION OF 264 00:09:39,500 --> 00:09:41,603 AGGREGATES WHICH WAS 1 OF THE 265 00:09:41,603 --> 00:09:42,537 HALLMARKS OF NEURODEGENERATION 266 00:09:42,537 --> 00:09:43,905 THAT I HIGHLIGHTED EARLIER. 267 00:09:43,905 --> 00:09:46,207 BUT WHAT IS EVEN MORE REMARKABLE 268 00:09:46,207 --> 00:09:49,410 ABOUT THIS RNA BINDING PROTEIN, 269 00:09:49,410 --> 00:09:54,048 IS THAT 97% OF ALS PATIENTS HAVE 270 00:09:54,048 --> 00:09:55,683 WILD-TYPE TDP 43 AGGREGATES. 271 00:09:55,683 --> 00:09:58,286 SO WHILE NORMALLY THIS PROTEIN 272 00:09:58,286 --> 00:10:00,955 IS DOICATE INDEED THE NUCLEOS -- 273 00:10:00,955 --> 00:10:04,592 LOCATED IN THE NUCLEUS, 97% OF 274 00:10:04,592 --> 00:10:06,427 PATIENTS EXHIBIT TPD43 IN THE 275 00:10:06,427 --> 00:10:07,762 CYTOPLASM THAT EXHIBIT THREEZ 276 00:10:07,762 --> 00:10:08,329 VARIOUS SHAPES HERE. 277 00:10:08,329 --> 00:10:12,166 SO BASED ON THIS PATHOLOGY, WE 278 00:10:12,166 --> 00:10:15,003 AND OTHERS THINK OF THIS 279 00:10:15,003 --> 00:10:16,971 PATHOLOGY BEING CAUSED BY A 280 00:10:16,971 --> 00:10:19,307 COMBINATION OF LOSS OF NUCLEAR 281 00:10:19,307 --> 00:10:22,343 TDP 43 FUNCTION AS WELL AS GAIN 282 00:10:22,343 --> 00:10:23,978 OF TOXIC CYTOPLASMIC FUNCTION. 283 00:10:23,978 --> 00:10:24,913 AND DEPENDING ON WHO YOU TALK 284 00:10:24,913 --> 00:10:27,315 TO, THERE WILL BE SOME THAT 285 00:10:27,315 --> 00:10:28,850 THINK IT'S THE LOSS OF NUCLEAR 286 00:10:28,850 --> 00:10:29,784 FUNCTION THAT MATTERS THAT WILL 287 00:10:29,784 --> 00:10:34,289 BE SOME THAT THINKS IT'S THE 288 00:10:34,289 --> 00:10:36,457 GAIN OF CYTOPLASMIC TOXIC 289 00:10:36,457 --> 00:10:38,660 FUNCTION THAT MATTERS AND AS 290 00:10:38,660 --> 00:10:39,827 TRUE IN LIFE, THE TRUTH IS 291 00:10:39,827 --> 00:10:40,762 SOMEWHERE IN BETWEEN. 292 00:10:40,762 --> 00:10:43,131 THE OTHER REMARKABLE THING ABOUT 293 00:10:43,131 --> 00:10:46,267 TDP43 IS THAT WHILE 97% OF ALS 294 00:10:46,267 --> 00:10:50,204 PATIENTS HAVE WILD-TYPE TDP 43 295 00:10:50,204 --> 00:10:52,407 AGGREGATES, 40% OF THE FRONTAL 296 00:10:52,407 --> 00:10:56,210 TEMPORAL PATIENTS ALSO HAVE 297 00:10:56,210 --> 00:10:57,345 WILD-TYPE TDP43 AGGREGATES. 298 00:10:57,345 --> 00:11:00,515 IN ADDITION ABOUT 50% OF 299 00:11:00,515 --> 00:11:02,617 ALZHEIMER'S PATIENTS EXHIBIT 300 00:11:02,617 --> 00:11:03,885 WILD-TYPE TDP43 AGGREGATE, 301 00:11:03,885 --> 00:11:05,353 INDIVIDUALS THAT SUFFERED 302 00:11:05,353 --> 00:11:09,524 TRAUMATIC BRAIN INJURY HAVE 303 00:11:09,524 --> 00:11:11,159 WILD-TYPE TDP 43 AGGREGATE, 304 00:11:11,159 --> 00:11:12,493 FOOTBALL PLAYERS WHO GET HIT A 305 00:11:12,493 --> 00:11:18,933 LOT WHO IS DIAGNOSE WIDE CET, 306 00:11:18,933 --> 00:11:19,734 HAVE WILD-TYPE AGGREGATES, SO IF 307 00:11:19,734 --> 00:11:22,103 YOU SEE IF WE STUDY THE BIOLOGY 308 00:11:22,103 --> 00:11:23,838 OF TDP43, HOW MIGHT THIS 309 00:11:23,838 --> 00:11:25,440 AGGREGATES COME ABOUT IN WHAT 310 00:11:25,440 --> 00:11:27,375 ARE THE PHENOTYPIC CONSEQUENCES 311 00:11:27,375 --> 00:11:28,109 OF THESE AGGREGATES? 312 00:11:28,109 --> 00:11:30,278 YOU MIGHT BE ABLE TO UNDERSTAND 313 00:11:30,278 --> 00:11:31,946 A LOT ABOUT MANY OF THESE 314 00:11:31,946 --> 00:11:33,314 DISORDERS THAT HAVE AS A COMMON 315 00:11:33,314 --> 00:11:37,752 DENATIONAL LIBRARY OF MEDICINE 316 00:11:37,752 --> 00:11:38,953 THEINATOR TDP43. 317 00:11:38,953 --> 00:11:42,023 SO AS TRAINED AS A FLIGHT 318 00:11:42,023 --> 00:11:43,391 GENETICIST DURING GRADUATE 319 00:11:43,391 --> 00:11:47,362 SCHOOL AND WORKING WITH FRAGILE 320 00:11:47,362 --> 00:11:50,999 X SYNDROME MODELS AS A POST DOC, 321 00:11:50,999 --> 00:11:52,066 I THOUGHT WHAT A GREAT 322 00:11:52,066 --> 00:11:53,134 OPPORTUNITY TO MODEL THIS IN 323 00:11:53,134 --> 00:11:54,202 FLIES AND I PROBABLY DON'T NEED 324 00:11:54,202 --> 00:11:55,636 TO SAY THIS TO MANY OF YOU IN 325 00:11:55,636 --> 00:11:57,505 THE AUDIENCE BUT I WILL SAY IT 326 00:11:57,505 --> 00:11:58,573 ANYWAY, FLIES ARE AWESOME, 327 00:11:58,573 --> 00:12:00,375 THERE'S A HIGH DEGREE OF 328 00:12:00,375 --> 00:12:02,377 CONSERVATION THAT ALLOWS US TO 329 00:12:02,377 --> 00:12:04,579 STUDY GENE FUNCTION AND IDENTIFY 330 00:12:04,579 --> 00:12:05,980 RELEVANT PATHWAYS TO HUMAN 331 00:12:05,980 --> 00:12:07,382 BIOLOGY AND HUMAN DISEASE IN THE 332 00:12:07,382 --> 00:12:09,417 FLY, THERE ARE MANY HUMAN 333 00:12:09,417 --> 00:12:11,285 DISEASE GENES THAT HAVE 334 00:12:11,285 --> 00:12:12,653 ORTHOLOGS IN THE FLY AND 335 00:12:12,653 --> 00:12:15,256 ALTHOUGH THERE ARE NOT IDENTICAL 336 00:12:15,256 --> 00:12:15,957 SYSTEMS, RIGHT? 337 00:12:15,957 --> 00:12:17,592 FLIES DON'T HAVE A LIVER PER SE 338 00:12:17,592 --> 00:12:19,827 BUT THEY HAVE A FAT BODY THAT 339 00:12:19,827 --> 00:12:21,396 PERFORMS MANY OF THE FUNCTIONS 340 00:12:21,396 --> 00:12:23,264 THE LIVER DOES, SO WHEN IT COMES 341 00:12:23,264 --> 00:12:25,366 TO THE NERVOUS SYSTEM, WE HAVE A 342 00:12:25,366 --> 00:12:26,667 PRETTY NICE SIMILARITY OR 343 00:12:26,667 --> 00:12:28,169 CORRELATION BETWEEN THE 2 344 00:12:28,169 --> 00:12:29,670 SYSTEMS, BOTH AT THE LEVEL OF 345 00:12:29,670 --> 00:12:31,305 CENTRAL NERVOUS SYSTEM AND 346 00:12:31,305 --> 00:12:33,274 PERIPHERAL NERVOUS SYSTEM, 347 00:12:33,274 --> 00:12:35,443 AGAIN, NOT IDENTICAL, BUT 348 00:12:35,443 --> 00:12:36,411 SUFFICIENT SIMILARITIES AND 349 00:12:36,411 --> 00:12:40,348 WONDERFUL GENETIC TOOLS THAT,A 350 00:12:40,348 --> 00:12:41,816 ALLOW US TO UNDERSTAND BASIC 351 00:12:41,816 --> 00:12:42,617 BIOLOGICAL QUESTIONS AND I DON'T 352 00:12:42,617 --> 00:12:46,154 NEED TO TELL YOU THAT THEY HAVE 353 00:12:46,154 --> 00:12:47,388 OTHER ADVANTAGES SUCH AS LOWER 354 00:12:47,388 --> 00:12:50,058 COST AND SHORT GENERATION TIME 355 00:12:50,058 --> 00:12:51,092 WHICH STUDENTS LIKE TO HEAR 356 00:12:51,092 --> 00:12:52,627 ABOUT BECAUSE THAT MEANS THEY 357 00:12:52,627 --> 00:12:54,328 WILL FINISH THEIR Ph.D. A 358 00:12:54,328 --> 00:12:55,430 LITTLE FASTER, RIGHT? 359 00:12:55,430 --> 00:12:58,733 THAT'S A GREAT SELLING POINT 360 00:12:58,733 --> 00:13:00,234 ANDIME THE ONLY FLIGHT 361 00:13:00,234 --> 00:13:01,436 GENETICIST IN MY COLLEGE, SO 362 00:13:01,436 --> 00:13:03,871 THAT'S BEEN A SUCCESSFUL 363 00:13:03,871 --> 00:13:05,239 RECRUITING TOOL. 364 00:13:05,239 --> 00:13:07,608 SO IN 2008, TDP 43 MUTATIONS 365 00:13:07,608 --> 00:13:08,709 WERE IDENTIFIED IN ALS AND 366 00:13:08,709 --> 00:13:12,246 THAT'S WHEN I DECIDED TO APPLY 367 00:13:12,246 --> 00:13:14,382 OUR SKILLS AND KNOWLEDGE OF 368 00:13:14,382 --> 00:13:15,450 FLIGHT IMENETTICS AND DISEASE 369 00:13:15,450 --> 00:13:17,585 MODELS AND MAKE FLY MODELS OF 370 00:13:17,585 --> 00:13:21,055 ALS, AND WE DECIDED TO DO THIS 371 00:13:21,055 --> 00:13:23,224 BY OVEREXPRESSING EITHER HUMAN 372 00:13:23,224 --> 00:13:26,828 TDP43 OR DO SOFILLA HOMOLOGUE OF 373 00:13:26,828 --> 00:13:29,497 TDP43 AND WE CAN DO IN THIS IN 374 00:13:29,497 --> 00:13:33,835 MOTOR NEURONS OR GLIAL CELLS OR 375 00:13:33,835 --> 00:13:35,269 MUSCLE CELLS, THOSE ARE ALL THE 376 00:13:35,269 --> 00:13:36,771 CELLS SKTAD IN THIS DISEASE. 377 00:13:36,771 --> 00:13:38,473 SO WE OVEREXPRESSED WILD-TYPE 378 00:13:38,473 --> 00:13:40,208 TDP43 AS A PROXY FOR THOSE 97% 379 00:13:40,208 --> 00:13:43,444 OF CASES WITH THE WILD-TYPE TDP 380 00:13:43,444 --> 00:13:45,012 PATHOLOGY, AND WE ALSO 381 00:13:45,012 --> 00:13:46,848 OVEREXPRESS VARIOUS MUTANT FORMS 382 00:13:46,848 --> 00:13:51,285 OF TDP43 AS A PROXY FOR TOAZ 383 00:13:51,285 --> 00:13:53,421 2-4% OF PATIENTS THAT EXHIBIT 384 00:13:53,421 --> 00:13:54,856 MUTATIONS IN TDP43. 385 00:13:54,856 --> 00:13:56,657 AND I'M SHOWING YOU HEAR, 386 00:13:56,657 --> 00:13:58,392 EXAMPLES OF PHENOTYPES THAT WE 387 00:13:58,392 --> 00:14:00,795 THINK ARE RELEVANT TO ALS, AND 388 00:14:00,795 --> 00:14:04,365 ALLOW US TO DO ALL KINDS OF 389 00:14:04,365 --> 00:14:05,066 PHENOTYPIC SCREENS INCLUDING 390 00:14:05,066 --> 00:14:06,501 DRUG SCREENS AND GENETIC SCREENS 391 00:14:06,501 --> 00:14:10,905 TO FIGURE OUT THE MECHANISTIC 392 00:14:10,905 --> 00:14:13,341 UNDERPINNINGS OF TDP43 PROTEIN 393 00:14:13,341 --> 00:14:14,609 OPEN MEETINGATHYS. 394 00:14:14,609 --> 00:14:17,678 SO OVEREXPRESS OF TDP43 MOTOR 395 00:14:17,678 --> 00:14:18,846 NEURONS CAUSES MOTOR NEURON 396 00:14:18,846 --> 00:14:21,048 DEATH, YOU CAN SEE FEWER 397 00:14:21,048 --> 00:14:23,217 MOTONEURONS IN THE THORACIC 398 00:14:23,217 --> 00:14:25,653 GANGLIA WHEN WE OVEREXPRESS 399 00:14:25,653 --> 00:14:28,456 HUMAN TDP 43, THERE ARE SYNAPTIC 400 00:14:28,456 --> 00:14:29,490 ABNORMALITIES OF THE 401 00:14:29,490 --> 00:14:30,191 NEUROMUSCULAR JUNCTION AND 402 00:14:30,191 --> 00:14:31,359 WITHOUT GOING INTO TOO MUCH 403 00:14:31,359 --> 00:14:33,995 DETAIL, I CAN TELL YOU THOSE 404 00:14:33,995 --> 00:14:35,296 SYNAPSES ARE ABNORMAL, THEY'RE 405 00:14:35,296 --> 00:14:38,900 SMALLER, SO THE SYNAPTIC 406 00:14:38,900 --> 00:14:40,535 DPLIEWRONS ARE THERE AND ALSO 407 00:14:40,535 --> 00:14:43,638 THERE ARE FUNCTIONAL DEFECTS IN 408 00:14:43,638 --> 00:14:45,139 SYNAPTIC VESICLE CYCLING, THE 409 00:14:45,139 --> 00:14:47,441 LIFE SPAN IS ALSO USED COMPARED 410 00:14:47,441 --> 00:14:49,777 TO WILD-TYPE FLY, WILD-TYPE 411 00:14:49,777 --> 00:14:51,245 TDP43 OVEREXPRESS CAUSES A 412 00:14:51,245 --> 00:14:53,748 REDUCTION IN LIFE SPAN AND 413 00:14:53,748 --> 00:14:55,816 MUTANT TDP43 OVEREXPRESS CAUSES 414 00:14:55,816 --> 00:14:57,251 AN EVEN MORE DRAMATIC AREY DUKS 415 00:14:57,251 --> 00:14:58,986 AND FOR AN ALS MODEL WE WANTED 416 00:14:58,986 --> 00:15:04,725 TO SEE WHETHER WE CAN HAVE OR 417 00:15:04,725 --> 00:15:06,694 DEVELOP ASSAYS THAT EXHIBIT 418 00:15:06,694 --> 00:15:08,329 OTHER HIGHLIGHT LOCOMOTOR 419 00:15:08,329 --> 00:15:09,997 DEFECTS CAUSED BY TDP43, AND 420 00:15:09,997 --> 00:15:11,632 THERE ARE MANY WAYS TO DO THIS 421 00:15:11,632 --> 00:15:14,302 IN THE FLY, MY FAVORITE ASSAY IS 422 00:15:14,302 --> 00:15:16,704 THE LARVA TURNING ASSAY BECAUSE 423 00:15:16,704 --> 00:15:18,339 IT'S QUICK, QUANTITATIVE, IT'S 424 00:15:18,339 --> 00:15:19,140 VISUALLY VERY SATISFYING AND 425 00:15:19,140 --> 00:15:21,309 COULD ALSO BE DONE BY 426 00:15:21,309 --> 00:15:22,243 UNDERGRADUATE STUDENTS WHICH 427 00:15:22,243 --> 00:15:24,412 WERE THE ENGINEER OF MY LAB WHEN 428 00:15:24,412 --> 00:15:26,013 I WAS AT THE UNIVERSITY OF 429 00:15:26,013 --> 00:15:26,247 ARIZONA. 430 00:15:26,247 --> 00:15:30,318 SO JUST TO GIVE YOU AN EXAMPLE, 431 00:15:30,318 --> 00:15:32,820 OVEREXPRESS OF WILD-TYPE TDP43 432 00:15:32,820 --> 00:15:36,791 NEURONS CAUSES A LARVA TURNING 433 00:15:36,791 --> 00:15:37,058 PHENOTYPE. 434 00:15:37,058 --> 00:15:40,094 SO WE FLIP THE LARVA AND THEN WE 435 00:15:40,094 --> 00:15:41,395 TAKE A TIMER AND COUNT LOW LONG 436 00:15:41,395 --> 00:15:43,998 IT TAKES FOR THIS ANIMAL TO TURN 437 00:15:43,998 --> 00:15:45,499 BACK AND START THE FIRST FORWARD 438 00:15:45,499 --> 00:15:47,034 MOVEMENT AND AFTER THIS GUY 439 00:15:47,034 --> 00:15:50,838 STOPS, I HOPE YOU CAN SEE THAT 440 00:15:50,838 --> 00:15:54,308 THIS CAUSES THE ANIMAL TO DRAG 441 00:15:54,308 --> 00:15:55,076 IT'S POSTERIOR END. 442 00:15:55,076 --> 00:15:58,012 SO I WILL START THIS AGAIN, AND 443 00:15:58,012 --> 00:16:05,886 I'M GOING TO ALSO START THE 1 ON 444 00:16:05,886 --> 00:16:08,422 THE RATE, THAT ATE WHAT WE CALL 445 00:16:08,422 --> 00:16:09,890 A PRODUCTIVE DRUG, SO I HOPE YOU 446 00:16:09,890 --> 00:16:11,826 CAN SEE THAT THE 1 ON THE RIGHT 447 00:16:11,826 --> 00:16:13,227 HAS ALREADY TURNED, WHILE THE 1 448 00:16:13,227 --> 00:16:16,631 ON THE LEFT THAT HASN'T EATEN, 449 00:16:16,631 --> 00:16:20,167 THE PROTECTIVE DRUG IS STILL 450 00:16:20,167 --> 00:16:20,568 STRUGGLING. 451 00:16:20,568 --> 00:16:23,271 SO WE USE THIS MODEL AND TRY TO 452 00:16:23,271 --> 00:16:25,473 UNDERSTAND ALS FOR MULTIPLE 453 00:16:25,473 --> 00:16:26,607 PERSECTIONAL ANALYSISSIVE USING 454 00:16:26,607 --> 00:16:28,709 GENETIC SCREENS EITHER UNBIASED 455 00:16:28,709 --> 00:16:31,912 OR CANDIDATE GENETIC SCREENS 456 00:16:31,912 --> 00:16:34,382 USING DRUG SCREENS, USING THE 457 00:16:34,382 --> 00:16:37,251 ALTERATIONS IN THIS ANIMAL, AND 458 00:16:37,251 --> 00:16:39,487 WHEN WE'RE THINKING ABOUT TDP 43 459 00:16:39,487 --> 00:16:41,522 AND GENERAL THINKING IN THE 460 00:16:41,522 --> 00:16:43,758 FIELD, AND I TOLD THAT YOU TDP43 461 00:16:43,758 --> 00:16:45,426 IS LOCATED IN THE NUCLEUS, 462 00:16:45,426 --> 00:16:46,827 HOWEVER THE CONTEXT OF DISEASE, 463 00:16:46,827 --> 00:16:53,134 IT LEAVES THE NUCLEUS, GOES INTO 464 00:16:53,134 --> 00:16:54,402 THE CYTOPLASM AND ASSOCIATES 465 00:16:54,402 --> 00:16:56,370 WITH VARIOUS COMPLEXES MOST OF 466 00:16:56,370 --> 00:16:57,471 WHICH CONDANE VARIOUS RNA 467 00:16:57,471 --> 00:16:58,773 BINDING PROTEINS AND AS 468 00:16:58,773 --> 00:17:00,041 AQUENCEQUENCE OF THAT, THERE ARE 469 00:17:00,041 --> 00:17:02,576 ALTERATIONS IN VARIOUS ASPECTS 470 00:17:02,576 --> 00:17:06,714 OF RNA METABOLISM, INCLUDING RNA 471 00:17:06,714 --> 00:17:07,481 TRANSLATION AND TRANSPORT. 472 00:17:07,481 --> 00:17:09,283 AND WHILE MANY LABS ARE FOCUSING 473 00:17:09,283 --> 00:17:12,620 ON THE NUCLEAR LOSS OF FUNCTION 474 00:17:12,620 --> 00:17:14,789 CONSEQUENCES OF TDP 43, 1 OF 475 00:17:14,789 --> 00:17:16,891 WHICH IS SPLICING DEFICITS, WE 476 00:17:16,891 --> 00:17:19,994 ARE FOCUSING MORE ON THE 477 00:17:19,994 --> 00:17:23,230 CYTOPLASMIC TOXIC ASPECTS OF TDP 478 00:17:23,230 --> 00:17:23,397 43. 479 00:17:23,397 --> 00:17:24,999 AND 1 OF THE HYPOTHESIS THAT HAS 480 00:17:24,999 --> 00:17:27,735 BEEN PUT FORTH IN THE FIELD BY 481 00:17:27,735 --> 00:17:30,137 COLLEAGUES MANY YEARS AGO, IS 482 00:17:30,137 --> 00:17:32,106 THAT ONCE IN THE CYTOPLASM, AND 483 00:17:32,106 --> 00:17:33,941 THAT CAN BE CAUSED BY STRESS 484 00:17:33,941 --> 00:17:38,245 WHETHER IT'S INJURY OR MUTATION 485 00:17:38,245 --> 00:17:39,647 OR AGING, TDP 43 CAN ASSOCIATE 486 00:17:39,647 --> 00:17:41,382 WITH OTHER RNA BINDING PROTEINS 487 00:17:41,382 --> 00:17:44,151 AND IN THE PROCESS CAN SEQUESTER 488 00:17:44,151 --> 00:17:46,654 THESE PROTEINS AND SEQUESTER 489 00:17:46,654 --> 00:17:48,322 COGNATE RNAs AND AS A 490 00:17:48,322 --> 00:17:49,690 CONSEQUENCE THOSE RNAs ARE NOT 491 00:17:49,690 --> 00:17:52,026 AVAILABLE TO THE TRANSLATIONAL 492 00:17:52,026 --> 00:17:57,098 MACHINERY FOR PROTEIN SYNTHESIS 493 00:17:57,098 --> 00:17:58,866 THIS WAS COINEDLET RIBOSTASIS 494 00:17:58,866 --> 00:18:00,234 HYPOTHESIS, AND IT MAKES A LOT 495 00:18:00,234 --> 00:18:02,403 OF SENSE, WE AND OTHERS HAVE 496 00:18:02,403 --> 00:18:03,804 BEEN TESTING THIS HYPOTHESIS AND 497 00:18:03,804 --> 00:18:06,874 I WILL SHOW YOU DATA THAT WE GOT 498 00:18:06,874 --> 00:18:08,743 FROM SPRPTS TESTING THESE -- 499 00:18:08,743 --> 00:18:09,677 EXPERIMENTS TESTING THESE 500 00:18:09,677 --> 00:18:10,678 HYPOTHESIS, BUT I WILL ALSO SAY 501 00:18:10,678 --> 00:18:15,783 THAT WHILE THIS ALL MAY SEEM 502 00:18:15,783 --> 00:18:17,151 REALLY INDIRECT, TDP43 MAY PLAY 503 00:18:17,151 --> 00:18:18,419 A MORE DIRECT ROLE IN 504 00:18:18,419 --> 00:18:20,054 TRANSLATION AND THAT IS 505 00:18:20,054 --> 00:18:22,923 SUGGESTED BY EVIDENCE THAT TDP43 506 00:18:22,923 --> 00:18:25,393 BINDING TO RAC 1, WHICH ALSO 507 00:18:25,393 --> 00:18:28,462 BINDS TO THE RIBOSOME, AND 508 00:18:28,462 --> 00:18:29,363 TDP43, WHICH ASSOCIATES WITH 509 00:18:29,363 --> 00:18:32,199 PROTEINS SO THERE MAY BE MORE TO 510 00:18:32,199 --> 00:18:33,601 TDP43'S ROLE IN TRANSLATION THAN 511 00:18:33,601 --> 00:18:35,503 THESE SORT OF SEQUESTRATION THAT 512 00:18:35,503 --> 00:18:38,606 CAN OCCUR IN THE CONTEXT OF 513 00:18:38,606 --> 00:18:39,039 DISEASE. 514 00:18:39,039 --> 00:18:40,040 TO TO ADDRESS THIS POSSIBILITY 515 00:18:40,040 --> 00:18:41,675 MANY YEARS AGO, FORMER GRADUATE 516 00:18:41,675 --> 00:18:44,678 STUDENT IN MY LAB, PERFORMED 517 00:18:44,678 --> 00:18:45,913 POLYSOME FRACTIONATIONS FROM 518 00:18:45,913 --> 00:18:48,449 FLIES OVER EXPRESSING WILD-TYPE 519 00:18:48,449 --> 00:18:49,817 OR MUTANT TDP 43OT RIGHT BUT 520 00:18:49,817 --> 00:18:51,685 ALSO FROM CONTROL FLIES AND ON 521 00:18:51,685 --> 00:18:53,921 THE LEFT IT CAN LOOK AT 522 00:18:53,921 --> 00:18:54,955 ENDOGENOUS TDP 43 AND YOU CAN 523 00:18:54,955 --> 00:18:57,324 SEE THAT IT BASICALLY 524 00:18:57,324 --> 00:18:58,959 DISTRIBUTES THROUGHOUT THE 525 00:18:58,959 --> 00:19:00,561 GRADIENT SUGGESTING THIS PROTEIN 526 00:19:00,561 --> 00:19:02,563 CAN HAVE A ROLE AS A NEGATIVE 527 00:19:02,563 --> 00:19:04,331 REGULATOR AS WELL AS POSITIVE 528 00:19:04,331 --> 00:19:09,236 REGULATOR OF TRANSLATION, AND 529 00:19:09,236 --> 00:19:11,505 THE OVEREXPRESSION OF HUMAN 530 00:19:11,505 --> 00:19:13,307 TDP43 EXHIBITED A SIMILAR 531 00:19:13,307 --> 00:19:16,477 PATTERN WITHIN THE POLYSOMES. 532 00:19:16,477 --> 00:19:19,880 TO FURTHER EXPLORE THE POTENTIAL 533 00:19:19,880 --> 00:19:21,482 RELATIONSHIP BETWEEN TDP43 AND 534 00:19:21,482 --> 00:19:23,751 TRANSLATION, 2 FORMER STUDENTS 535 00:19:23,751 --> 00:19:25,386 IN MY LAB, WHILE AT THE 536 00:19:25,386 --> 00:19:27,555 UNIVERSITY OF ARIZONA USE 537 00:19:27,555 --> 00:19:29,490 GENETIC INTERACTIONS TO MODULATE 538 00:19:29,490 --> 00:19:31,592 TRANSLATION, EITHER BY KNOCKING 539 00:19:31,592 --> 00:19:35,329 DOWN 40 DP WHICH IS AN INHIBITOR 540 00:19:35,329 --> 00:19:38,833 OF INITIATION OR OVEREXPRESSING 541 00:19:38,833 --> 00:19:45,172 THE CONSTITTATIVELY ACTIVE F6 542 00:19:45,172 --> 00:19:46,774 KINASE OF THE LEVEL OF 543 00:19:46,774 --> 00:19:47,741 QUANTIFICATION. 544 00:19:47,741 --> 00:19:50,578 SO WHAT YOU'RE LOOKING AT IS THE 545 00:19:50,578 --> 00:19:52,646 LEVEL OF ASSAY, I SHOWED YOU 546 00:19:52,646 --> 00:19:55,182 EARLIER, THE TAKE IS LONGER THAN 547 00:19:55,182 --> 00:19:57,952 CONTROLS AND THIS IS RESCUED BY 548 00:19:57,952 --> 00:20:05,626 KNOCKING DOWN FOUR BT OR 549 00:20:05,626 --> 00:20:06,393 CONSTITTATIVELY ASSESSING 550 00:20:06,393 --> 00:20:07,027 KINASE, CONSISTENT ALTOGETHER 551 00:20:07,027 --> 00:20:08,562 WITH THE FACT THAT THE NOTION 552 00:20:08,562 --> 00:20:11,632 THAT IF YOU PROMOTE TRANSLATION, 553 00:20:11,632 --> 00:20:14,301 YOU CAN MITIGATE TDP43 554 00:20:14,301 --> 00:20:15,069 ASSOCIATED LOCOMOTOR DEFECTS. 555 00:20:15,069 --> 00:20:16,770 OTHER WAYS IN WHICH WE'RE 556 00:20:16,770 --> 00:20:19,073 STUDYING THE ROLE, OR THE 557 00:20:19,073 --> 00:20:20,574 CONNECTION, INVOLVEMENT OF TD43 558 00:20:20,574 --> 00:20:23,611 IN TRANSLATION, IS USING 559 00:20:23,611 --> 00:20:26,313 GENETICALLY ENCODED APPROACH 560 00:20:26,313 --> 00:20:28,916 CALLED NONCANNONICLE TAGGING 561 00:20:28,916 --> 00:20:30,217 DEVELOPED IN SHYOUMAN'S LAB 562 00:20:30,217 --> 00:20:32,820 WHERE WE BASICALLY OVEREXPRESS A 563 00:20:32,820 --> 00:20:34,955 MUTANT FORM OF TRNA SYNTHETASE 564 00:20:34,955 --> 00:20:39,560 AND THIS MUTANT FORM OF 565 00:20:39,560 --> 00:20:40,928 METHIONINE SYNTHETASE CANNOT 566 00:20:40,928 --> 00:20:42,563 CHARGE THE IPADDA 9 ON TO NEW 567 00:20:42,563 --> 00:20:45,232 PEPTIDES BUT CAN DO SO WITH 568 00:20:45,232 --> 00:20:47,067 AN ANALOGUE WHICH IS CALLED ANL, 569 00:20:47,067 --> 00:20:49,837 AND IF WE FEED ANL IN THE FEED 570 00:20:49,837 --> 00:20:52,039 THEN WE OVEREXPRESS IT IN THE 571 00:20:52,039 --> 00:20:54,675 RNA SYNTHETASE IN MOTOR NEURONS, 572 00:20:54,675 --> 00:20:57,144 ALL THE NEWLY SIPGHT SIZED 573 00:20:57,144 --> 00:20:59,079 PROTEINS NEURONS WILL NOW HAVE 574 00:20:59,079 --> 00:20:59,880 NEWEL MADE PROTEINS STARTING 575 00:20:59,880 --> 00:21:01,582 WITH A, IN L AND THE NICE THING 576 00:21:01,582 --> 00:21:05,119 ABOUT ANL IS THAT IT CAN BE 577 00:21:05,119 --> 00:21:06,253 LABELED USING CLICK CHEMISTRY 578 00:21:06,253 --> 00:21:09,323 OTHERS EITHER WITH FLUORESCENT 579 00:21:09,323 --> 00:21:12,159 MARKERS AND WE CALL THAT 580 00:21:12,159 --> 00:21:15,229 APPROACH FUN CAT OR CAN BE 581 00:21:15,229 --> 00:21:19,500 LABELED WITH BIOTIN AND CALL IT 582 00:21:19,500 --> 00:21:22,670 BBONCAT, AND TURNS OUT WE ARE 583 00:21:22,670 --> 00:21:25,773 CAT LOVERS SO WE USE FUNCAT AND 584 00:21:25,773 --> 00:21:28,509 BONCAT, TO STUDY THESE ON NEWLY 585 00:21:28,509 --> 00:21:28,976 SYNTHITIZED PROTEINS. 586 00:21:28,976 --> 00:21:31,245 AND I HOPE THAT WHAT YOU CAN SEE 587 00:21:31,245 --> 00:21:37,284 HERE, IS THAT AT THE NEURAL 588 00:21:37,284 --> 00:21:39,653 MUSCULAR JUNCTION, THE 589 00:21:39,653 --> 00:21:41,455 FLUORESCENT LABEL OF ANL PROTEIN 590 00:21:41,455 --> 00:21:44,091 SYSTEM REDUCED COMPARED TO 591 00:21:44,091 --> 00:21:44,758 CONTROLS. 592 00:21:44,758 --> 00:21:47,428 WITH THE BONCAT APPROACH, WE CAN 593 00:21:47,428 --> 00:21:50,831 PULL DOWN THE BIOATTENUATE 594 00:21:50,831 --> 00:21:52,399 ANL PROTEIN AND DO MASS 595 00:21:52,399 --> 00:21:53,934 SPECTROMETRY AND IERK DENTIFY 596 00:21:53,934 --> 00:21:55,669 SPECIFIC CHANGES IN DOSE 597 00:21:55,669 --> 00:21:57,605 PROTEINS THAT ARE NOT 598 00:21:57,605 --> 00:21:59,473 SYNTHESIZED APPROPRIATELY IN THE 599 00:21:59,473 --> 00:22:00,541 CONTEXT OF TDP 43. 600 00:22:00,541 --> 00:22:02,409 THEN IN MOTOR NEURONS, THE 601 00:22:02,409 --> 00:22:03,277 EXPERIMENTS ARE IN PROGRESS AND 602 00:22:03,277 --> 00:22:04,878 THERE ARE A LITTLE BIT MORE 603 00:22:04,878 --> 00:22:06,180 CHALLENGING BECAUSE, THERE ARE 604 00:22:06,180 --> 00:22:07,681 FEW MOTOR NEURONS AND THE SIGNAL 605 00:22:07,681 --> 00:22:09,149 TO NOISE RATIO IS CHALLENGING 606 00:22:09,149 --> 00:22:10,084 BUT WE'RE WORKING ON IT. 607 00:22:10,084 --> 00:22:13,053 AND I WILL SHOW YOU A LITTLE BIT 608 00:22:13,053 --> 00:22:15,589 LATER DATA USING THIS,A PROACH 609 00:22:15,589 --> 00:22:16,857 IN MUSCLE, WHERE -- THIS 610 00:22:16,857 --> 00:22:18,225 APPROACH IN MUSCLE WHERE THE 611 00:22:18,225 --> 00:22:21,195 DEFECT IS VERY CLEAR. 612 00:22:21,195 --> 00:22:23,097 SO, THINKING ABOUT TRANSLATION, 613 00:22:23,097 --> 00:22:24,898 THINKING ABOUT RNAs THAT 614 00:22:24,898 --> 00:22:27,001 ASSOCIATE WITH TDP 43, AND ARE 615 00:22:27,001 --> 00:22:30,004 DISREGULATED IN THE CONTEXT OF 616 00:22:30,004 --> 00:22:32,373 TDP 43 PROTEIN OP A THY, WE OVER 617 00:22:32,373 --> 00:22:34,041 THE YEARS WE IDENTIFY A NUMBER 618 00:22:34,041 --> 00:22:36,010 OF RNAs THAT ARE ASSOCIATE 619 00:22:36,010 --> 00:22:38,078 WIDE TDP43 AND DISREGULATED IN 620 00:22:38,078 --> 00:22:40,547 MOTOR NEURONS IN THE CONTEXT OF 621 00:22:40,547 --> 00:22:42,549 TDP43, OUR VERY FIRST TARGET WE 622 00:22:42,549 --> 00:22:44,652 IDENTIFIED WAS THE CYTOSKELETAL 623 00:22:44,652 --> 00:22:46,720 COMPONENT CALLED FORTUNE FLIES, 624 00:22:46,720 --> 00:22:48,822 IS THE ORTHOLOG OF MAP 1 B IN 625 00:22:48,822 --> 00:22:51,125 HUMANS WHICH IS A MICROTUBEULAR 626 00:22:51,125 --> 00:22:55,863 ASSOCIATED PROTEIN THAT 627 00:22:55,863 --> 00:23:01,935 STABILIZES PROTEINS AND AS YOU 628 00:23:01,935 --> 00:23:03,637 CAN SEE USING THIS NEUROMUSCULAR 629 00:23:03,637 --> 00:23:05,506 SYSTEM IN THE FLY, WE WERE ABLE 630 00:23:05,506 --> 00:23:07,241 TO STUDY PROTEIN GENE EXPRESSION 631 00:23:07,241 --> 00:23:08,809 IN THE VENTRAL CORD WITH ALL THE 632 00:23:08,809 --> 00:23:10,544 MOTOR NEURONS THAT ARE 633 00:23:10,544 --> 00:23:11,845 ALLOCATED, WE CAN LOOK AT THE 634 00:23:11,845 --> 00:23:13,614 AXON, WE CAN LOOK AT THE 635 00:23:13,614 --> 00:23:16,150 SYNAPTIC TERMINAL SO USING THE 636 00:23:16,150 --> 00:23:20,087 SYSTEM, WE FOUND THAT PROTEIN, 637 00:23:20,087 --> 00:23:22,589 IS REDUCED AT THE SYNAPTIC 638 00:23:22,589 --> 00:23:24,158 TERMINAL BUT IS INCREASED IN THE 639 00:23:24,158 --> 00:23:26,026 MOTOR NEURON CELL BODY AND IN 640 00:23:26,026 --> 00:23:27,561 THE VENTRAL QUARTER AND THIS IS 641 00:23:27,561 --> 00:23:30,164 QUANTIFIED HERE AND THIS IS 642 00:23:30,164 --> 00:23:31,465 IMMUNO FLUORESCENCE OF FOOT 643 00:23:31,465 --> 00:23:33,834 PROTEIN IN THE TDP43 EXPRESSING 644 00:23:33,834 --> 00:23:37,571 ANIMAL, AND THAT WAS THE VERY 645 00:23:37,571 --> 00:23:38,739 FIRST TIME WHEN WHICH WAS 646 00:23:38,739 --> 00:23:40,240 BROUGHT TO MY ATTENTION THAT 647 00:23:40,240 --> 00:23:42,176 THIS IS ALL NICE AND INTERESTING 648 00:23:42,176 --> 00:23:43,711 BUT WHAT DOES THIS MEAN FOR THE 649 00:23:43,711 --> 00:23:45,612 HUMAN DISEASE AND THE CLINICIAN 650 00:23:45,612 --> 00:23:46,180 TOLD ME THAT. 651 00:23:46,180 --> 00:23:48,615 I SAID OKAY, THATIA A GOOD POINT 652 00:23:48,615 --> 00:23:50,951 SO THAT'S WHEN WE STARTED 653 00:23:50,951 --> 00:23:51,585 COLLABORATIONS WITH COLLEAGUES 654 00:23:51,585 --> 00:23:53,153 IN THE FIELD THAT WE'RE WORKING 655 00:23:53,153 --> 00:23:54,755 WITH PATIENT TISSUES AND WE 656 00:23:54,755 --> 00:23:55,923 STARTED LOOKING AT PATIENT 657 00:23:55,923 --> 00:23:57,591 TISSUES OURSELVES, TO VALIDATE 658 00:23:57,591 --> 00:23:59,126 THE TARGETS WITH WHETHER THE 659 00:23:59,126 --> 00:24:00,360 TARGETS WERE IEE, AUDIENCE 660 00:24:00,360 --> 00:24:01,795 DENTIFIED IN THE FLY OR ALSO 661 00:24:01,795 --> 00:24:03,764 ALTERED IN THE CONTEXT OF HUMAN 662 00:24:03,764 --> 00:24:04,732 DEC TO ULTIMATELY TEST THE 663 00:24:04,732 --> 00:24:06,834 RELEVANCE OF OUR WORK TO ALS. 664 00:24:06,834 --> 00:24:08,802 AND I HOPE YOU CAN SEE HERE, 665 00:24:08,802 --> 00:24:11,305 THAT IN PATIENTS SPINAL CORDS, 666 00:24:11,305 --> 00:24:14,208 MAP 1 B, IS ALSO UPREGULATED IN 667 00:24:14,208 --> 00:24:17,644 MOTOR NEURON CELL BODIES AS WE 668 00:24:17,644 --> 00:24:21,582 IDENTIFIED IT IN THE FLY. 669 00:24:21,582 --> 00:24:23,283 AND THIS WAS INTERESTING AT THE 670 00:24:23,283 --> 00:24:26,220 TIME BECAUSE SINGLE CELL RNA SEQ 671 00:24:26,220 --> 00:24:27,387 WASN'T AVAILABLE, RIGHT, LIKE 672 00:24:27,387 --> 00:24:29,123 THE IDENTIFICATION OF TARGETS IN 673 00:24:29,123 --> 00:24:30,624 HUMAN TISSUES WAS MUCH HARDER SO 674 00:24:30,624 --> 00:24:33,227 HAVING THE ABILITY TO DO THIS IN 675 00:24:33,227 --> 00:24:37,564 THE FLY WAS REALLY HELPFUL IN 676 00:24:37,564 --> 00:24:46,707 IDENTIFYING MARKERS OF PATHOLOGY 677 00:24:46,707 --> 00:24:48,075 IN PATIENT TISSUES. 678 00:24:48,075 --> 00:24:53,380 SOON AFTERRER WE FOUND A VESICLE 679 00:24:53,380 --> 00:24:55,249 CALLED HSP8, AND WE FOUND IN 680 00:24:55,249 --> 00:24:57,451 FLIES AS WELL AS PATIENT DERIVED 681 00:24:57,451 --> 00:24:59,086 NEURONS THE EXPRESSION OF THE 682 00:24:59,086 --> 00:25:00,454 SYNAPTIC TERMINAL WAS REDUCED 683 00:25:00,454 --> 00:25:03,223 OVER EXPRESS OF IT IN THE FLY 684 00:25:03,223 --> 00:25:05,225 RESCUED ALL THE PHENOTYPES 685 00:25:05,225 --> 00:25:06,727 CAUSED BY TDP43, AND THIS IS 686 00:25:06,727 --> 00:25:09,897 WHEN WE STARTED EXPANDING FROM 687 00:25:09,897 --> 00:25:11,999 LOOKING AT TDP43 PROTEIN OP A 688 00:25:11,999 --> 00:25:22,543 THY TO OTHER TYPES OF ALS, THAT 689 00:25:22,876 --> 00:25:23,610 EXHIBIT THE PATHOLOGY. 690 00:25:23,610 --> 00:25:27,447 SO WE'VE BEGUN TO EXPAND OUR 691 00:25:27,447 --> 00:25:29,116 ABILITY TO SWIR ANSWER QUESTIONS 692 00:25:29,116 --> 00:25:31,351 AND UNDERSTAND OTHER TYPES OF 693 00:25:31,351 --> 00:25:33,187 ALS BY USING THESE OTHER TYPES 694 00:25:33,187 --> 00:25:35,989 OF MODELS AND INTERESTINGLY 695 00:25:35,989 --> 00:25:37,591 ENOUGH, CHAPERON WAS ALSO 696 00:25:37,591 --> 00:25:39,226 ALTERED IN THE FLY MODELS AND 697 00:25:39,226 --> 00:25:40,761 THIS IS AN EXPERIMENT WHERE 698 00:25:40,761 --> 00:25:46,733 LOOKING AT THE UPTAKE OF TDP43, 699 00:25:46,733 --> 00:25:48,268 AND YOU CAN SEE THIS IS REDUCED 700 00:25:48,268 --> 00:25:50,671 IN THE CONTEXT OF THE MODEL BUT 701 00:25:50,671 --> 00:25:54,608 OVER EXPRESS OF HSC7-4 RESCUED 702 00:25:54,608 --> 00:25:55,108 THE VESICLE CYCLE. 703 00:25:55,108 --> 00:25:56,710 SO THIS WAS A TYPE OF APPROACH 704 00:25:56,710 --> 00:25:59,680 THAT WE USE LIKE SORT OF GOING 1 705 00:25:59,680 --> 00:26:02,449 TARGET AT A TIME, IDENTIFYING 706 00:26:02,449 --> 00:26:04,318 WHAT IS TDP43, REGULATING AND 707 00:26:04,318 --> 00:26:07,754 WHAT DO THESE TARGETS -- HOW DO 708 00:26:07,754 --> 00:26:10,057 THEY MEDIATE THE TOXICITY OF 709 00:26:10,057 --> 00:26:11,892 TDP43, DO THEY AFFECT MOTOR 710 00:26:11,892 --> 00:26:13,627 FUNCTION, DO THEY AFFECT 711 00:26:13,627 --> 00:26:15,128 DEGENERATION, DO THEY AFFECT 712 00:26:15,128 --> 00:26:16,330 SYNAPTIC FUNCTION, SO THE NEXT 713 00:26:16,330 --> 00:26:23,937 TARGET WE WORKED ON IS THIS 714 00:26:23,937 --> 00:26:26,340 DALLY LIKE PROTEIN, IT PLAYS AN 715 00:26:26,340 --> 00:26:27,407 IMPORTANT PROTEIN IN WIN 716 00:26:27,407 --> 00:26:31,645 SIGNALING AND IN HUMANS 1 OF THE 717 00:26:31,645 --> 00:26:33,480 ORTHOLOGS WAS GPC 6, AND WHAT WE 718 00:26:33,480 --> 00:26:36,116 FOUND IS THAT AT THE SYNAPTIC 719 00:26:36,116 --> 00:26:37,784 TERMINAL THIS PROTEIN IS REDUCED 720 00:26:37,784 --> 00:26:38,118 IN EXPRESSION. 721 00:26:38,118 --> 00:26:40,254 THIS TIME WE ALSO STARTED 722 00:26:40,254 --> 00:26:41,622 LOOKING AT TDP43 LOSS OF 723 00:26:41,622 --> 00:26:43,690 FUNCTION TO REALLY COMPARE THE 724 00:26:43,690 --> 00:26:44,725 OVEREXPRESSION MODEL WITH THE 725 00:26:44,725 --> 00:26:45,859 LOSS OF FUNCTION MODELS, AND I 726 00:26:45,859 --> 00:26:48,061 HOPE YOU CAN SEE THAT THE 727 00:26:48,061 --> 00:26:49,463 SYNAPTIC TERMINAL WE HAVE A 728 00:26:49,463 --> 00:26:52,432 SIMILAR EFFECT, A SLIGHT BUT 729 00:26:52,432 --> 00:26:53,467 STATISTICALLY SIGNIFICANT 730 00:26:53,467 --> 00:26:55,202 REDUCTION IN DLP EXPRESSION, BUT 731 00:26:55,202 --> 00:26:57,337 TO OUR SURPRISE, THERE WAS A 732 00:26:57,337 --> 00:26:58,005 COMPARTMENT SPECIFIC PHENOTYPE, 733 00:26:58,005 --> 00:26:59,439 WHEN WE LOOKED IN THE VENTRAL 734 00:26:59,439 --> 00:27:02,075 CORD WHERE THE MOTOR NEURONS ARE 735 00:27:02,075 --> 00:27:06,046 LOCATED WE FOUND THAT DLP WAS 736 00:27:06,046 --> 00:27:06,947 FORMING THESE ACCUMULATIONS THAT 737 00:27:06,947 --> 00:27:10,083 WERE SPECIFIC TO THE TDP43 738 00:27:10,083 --> 00:27:11,885 OVEREXPRESSION, THEY ARE NOT 739 00:27:11,885 --> 00:27:12,819 PRESENT IN THE CONTROL, AND 740 00:27:12,819 --> 00:27:13,687 THEY'RE SORT OF MIDDLE OF THE 741 00:27:13,687 --> 00:27:18,492 ROAD IN THE LOSS OF FUNCTION. 742 00:27:18,492 --> 00:27:20,928 AND SO, AGAIN WOORKSY WANTED TO 743 00:27:20,928 --> 00:27:23,664 KNOW WHETHER GPC 6 IS ALTERED IN 744 00:27:23,664 --> 00:27:27,534 PATIENT TISSUES, AND LOOKING AT 745 00:27:27,534 --> 00:27:30,103 SPINAL CORDS FROM ALS CASES OF 746 00:27:30,103 --> 00:27:32,639 TDP43 PATHOLOGY, WE FOUND THERE 747 00:27:32,639 --> 00:27:34,174 IS ACCUMULATION OF GPC 6 TARGET 748 00:27:34,174 --> 00:27:36,810 THAT IS QUANTIFIED OVER HERE IN 749 00:27:36,810 --> 00:27:38,679 PATIENT TISSUE, ONCE AGAIN 750 00:27:38,679 --> 00:27:39,680 HIGHLIGHTING THE POWER, THE 751 00:27:39,680 --> 00:27:41,949 PREDICTIVE POWER OF THE FLY, OUR 752 00:27:41,949 --> 00:27:43,250 ABILITY TO IDENTIFY MRNA TARGETS 753 00:27:43,250 --> 00:27:44,985 THAT ARE ALTERED IN FLIES BUT 754 00:27:44,985 --> 00:27:46,653 ALSO IN PATIENT TISSUES. 755 00:27:46,653 --> 00:27:47,587 AND WE'RE CURRENTLY TRYING TO 756 00:27:47,587 --> 00:27:50,624 FIGURE OUT WHAT IS THIS 757 00:27:50,624 --> 00:27:51,758 COMPARTMENT SPECIFIC 758 00:27:51,758 --> 00:27:52,960 DISREGULATION OF DLP MEAN, SO 759 00:27:52,960 --> 00:27:54,461 FOR EXAMPLE, WE SEE THESE 760 00:27:54,461 --> 00:27:59,466 ACCUMULATION IN THE CELL BODY, 761 00:27:59,466 --> 00:28:00,233 DOING MEMBRANE FRACTIONATIONS, 762 00:28:00,233 --> 00:28:03,270 WE FOUND THAT THE ENDOPLASMIC 763 00:28:03,270 --> 00:28:04,905 RETIC LUMIS FRAGMENTED IN THE 764 00:28:04,905 --> 00:28:07,641 CONTEXT, THERE ARE INTRACELLULAR 765 00:28:07,641 --> 00:28:09,710 TRANSPORT DEFICITS THAT CAUSE 766 00:28:09,710 --> 00:28:11,244 FRAGMENTATION IN THE ENDOPLAZ 767 00:28:11,244 --> 00:28:13,046 MIDSIC RETIC LUMAND SURELY 768 00:28:13,046 --> 00:28:14,081 ENOUGH WHILE DLPs ARE 769 00:28:14,081 --> 00:28:15,382 ASSOCIATE WIDE THE CONTROL, YOU 770 00:28:15,382 --> 00:28:19,853 CAN SEE IT'S KIND OF ENRICHED IN 771 00:28:19,853 --> 00:28:20,988 THE FRAGMENTED ENDOPLASMIC RETIC 772 00:28:20,988 --> 00:28:23,757 LUMTHAT WE FIND IN THE CONTEXT 773 00:28:23,757 --> 00:28:26,860 OF TDP43 MUTANT MODEL AND THE 774 00:28:26,860 --> 00:28:29,262 SYNAPTIC TERMINAL, THIS IS SUPER 775 00:28:29,262 --> 00:28:32,766 INTERESTING BECAUSE DLP IN 776 00:28:32,766 --> 00:28:33,433 RESPONSE TO CCLSTIMMULATION GOES 777 00:28:33,433 --> 00:28:36,770 TO THE MEMBRANE AND THAT'S WHAT 778 00:28:36,770 --> 00:28:40,374 IT SIGNALS TO WINLESS RECEPTORS 779 00:28:40,374 --> 00:28:41,208 AND POTENTIALLY TDMP RECEPTORS 780 00:28:41,208 --> 00:28:43,210 AND WE FIND THAT WHILE DLP CAN 781 00:28:43,210 --> 00:28:45,078 DO THAT IN THE CONTEXT OF THE 782 00:28:45,078 --> 00:28:47,247 MUTANT, THERE IS MUCH LESS OF IT 783 00:28:47,247 --> 00:28:50,584 BECAUSE OF THE EFFECT OF TDP43 784 00:28:50,584 --> 00:28:52,886 ON THE OVERALL EXPRESS OF THE 785 00:28:52,886 --> 00:28:54,621 SYNAPTIC TERMINAL, SO WE 786 00:28:54,621 --> 00:28:55,889 HYPOTHESIZED THIS HAS AN EFFECT 787 00:28:55,889 --> 00:28:58,792 ON THE WINLESS SIGNALING AT THE 788 00:28:58,792 --> 00:28:59,693 NEURAL MUSCULAR JUNCTION IS THIS 789 00:28:59,693 --> 00:29:00,861 IS SOMETHING WE'RE CURRENTLY 790 00:29:00,861 --> 00:29:01,928 WORKING ON. 791 00:29:01,928 --> 00:29:03,563 SO DURING THIS 1 TARGET OF TIME 792 00:29:03,563 --> 00:29:04,431 IS INTERESTING BUT WE ALSO 793 00:29:04,431 --> 00:29:07,367 WANTED TO HAVE A BETTER 794 00:29:07,367 --> 00:29:09,436 UNDERSTANDING OF ALL THE TARGETS 795 00:29:09,436 --> 00:29:11,438 THAT TDP43 MAY REGULATE IN MOTOR 796 00:29:11,438 --> 00:29:14,541 NEURONS AND SO 2 OF MY STUDENTS 797 00:29:14,541 --> 00:29:17,577 DECIDED TO USE RIBOTAGGING TO 798 00:29:17,577 --> 00:29:19,446 IDENTIFY ALL THE MRNAs THAT DO 799 00:29:19,446 --> 00:29:21,081 NOT ASSOCIATE WITH RIBOSOMES 800 00:29:21,081 --> 00:29:23,984 NORMAL IN THE CONTEXT OF TDP43, 801 00:29:23,984 --> 00:29:27,754 SO THESE ARE JUST VISUALIZATION 802 00:29:27,754 --> 00:29:29,723 OF DATA WE GOT THROUGH 803 00:29:29,723 --> 00:29:31,291 RIBOTAGGING SHOWING YOU THAT THE 804 00:29:31,291 --> 00:29:33,627 RNATHAT IS HAVE DIFFERENT LEVELS 805 00:29:33,627 --> 00:29:35,395 OF ASSOCIATION WITH RIBOSOMES IN 806 00:29:35,395 --> 00:29:36,797 METEOR NEURONS SPECIFICALLY WHEN 807 00:29:36,797 --> 00:29:41,334 WE OVEREXPRESS THE TDP43, 808 00:29:41,334 --> 00:29:44,404 FALLING TO SPLICING IF YOU DO A 809 00:29:44,404 --> 00:29:46,139 TERM ANALYSIS, MANY ARE 810 00:29:46,139 --> 00:29:48,075 ASSOCIATE WIDE SPLICING BUT 811 00:29:48,075 --> 00:29:49,009 WHAT'S DIFFICULT TO FIGURE OUT 812 00:29:49,009 --> 00:29:51,812 HERE IS YOU SEE MORE ARE MORE 813 00:29:51,812 --> 00:29:52,946 ASSOCIATED WITH RIBOSOMES AND 814 00:29:52,946 --> 00:29:54,181 SOME ARE LESS ASSOCIATE WIDE 815 00:29:54,181 --> 00:29:54,681 HYPOSTUDIES OF MULTIPLE 816 00:29:54,681 --> 00:29:55,515 ENDOCRINES SO IT'S DIFFICULT TO 817 00:29:55,515 --> 00:29:56,917 MAKE SENSE OF WHAT'S GOING ON IN 818 00:29:56,917 --> 00:29:57,918 REGARD TO THIS PATHWAY. 819 00:29:57,918 --> 00:30:01,088 WHEN WE LOOK THEA ELECTRON 820 00:30:01,088 --> 00:30:04,458 TRANSPORT, YOU CAN SEE THAT 821 00:30:04,458 --> 00:30:06,827 COMPLEX 1 COMPONENT TRANSCRIPT, 822 00:30:06,827 --> 00:30:07,894 ACTUALLY MORE ASSOCIATE WIDE 823 00:30:07,894 --> 00:30:10,030 RIBOSOME IN THE CONTEXT OF 824 00:30:10,030 --> 00:30:10,230 TDP43. 825 00:30:10,230 --> 00:30:12,999 AND WE THINK THAT THAT'S 826 00:30:12,999 --> 00:30:13,834 ACTUALLY A COMPENSATORY 827 00:30:13,834 --> 00:30:16,002 MECHANISM BECAUSE OF DEFICITS IN 828 00:30:16,002 --> 00:30:18,672 MITOCHONDRIA, THERE IS THE 829 00:30:18,672 --> 00:30:20,140 COMPENSATORY MECHANISM, TRYING 830 00:30:20,140 --> 00:30:21,675 TO MAKE MORE OF COMPLEX 1 831 00:30:21,675 --> 00:30:24,311 PROTEIN AND TRY TO RESTORE 832 00:30:24,311 --> 00:30:25,145 MITOCHONDRIA FUNCTION IN THIS 833 00:30:25,145 --> 00:30:26,880 WAY, BUT WHAT WAS A LITTLE BIT 834 00:30:26,880 --> 00:30:30,684 MORE CLEAR, AND EASY TO APPROACH 835 00:30:30,684 --> 00:30:33,787 WAS A DOWN REGULATION OF THE 836 00:30:33,787 --> 00:30:36,456 TRANSCRIPTS INVOLVED IN 837 00:30:36,456 --> 00:30:36,957 TRANSLATION INITIATION. 838 00:30:36,957 --> 00:30:39,693 SO YOU CAN SEE THAT MANY OF 839 00:30:39,693 --> 00:30:40,994 THESE ARE LESS ASSOCIATE WIDE 840 00:30:40,994 --> 00:30:48,468 RIEB SOSOMES IN THE CONTEXT OF 841 00:30:48,468 --> 00:30:49,469 TDP43 CONSISTENT WITH TDP43 842 00:30:49,469 --> 00:30:51,404 OVEREXPRESSION HAVING AN EFFECT 843 00:30:51,404 --> 00:30:54,374 ON TRANSLATION AND INITIATION IN 844 00:30:54,374 --> 00:30:55,208 GENERAL. 845 00:30:55,208 --> 00:30:57,878 AND 1 OF THE OTHER INTERESTING 846 00:30:57,878 --> 00:30:59,112 PATHWAYS THAT SHOWED A VERY 847 00:30:59,112 --> 00:31:04,084 CLEAR EFFECT FROM THE 848 00:31:04,084 --> 00:31:05,585 RIBOTAGGING EXPERIMENT, IS THE 849 00:31:05,585 --> 00:31:06,119 PHOSPHATE PATHWAY. 850 00:31:06,119 --> 00:31:09,923 THIS IS SORT OF AN ARMADILLOS OF 851 00:31:09,923 --> 00:31:11,658 GLYCOLYSIS, THAT NEURONS USE AND 852 00:31:11,658 --> 00:31:14,628 CELLS USE, WHEN THEY ARE 853 00:31:14,628 --> 00:31:16,429 UNDERSTRESS, AND IT WAS REALLY 854 00:31:16,429 --> 00:31:20,834 INTERESTING TO SEE THAT THE 855 00:31:20,834 --> 00:31:23,436 COMPONENTS OF THE PENTINE 856 00:31:23,436 --> 00:31:25,672 REGIMEN OS PHOSPHATE PATHWAY 857 00:31:25,672 --> 00:31:27,541 WERE LESS PREDICTIVE. 858 00:31:27,541 --> 00:31:29,309 THEY WERE LESS RIBOSOME 859 00:31:29,309 --> 00:31:31,311 INCLUDING ZEST WHITE WHICH IS IN 860 00:31:31,311 --> 00:31:34,514 FLIES THE RATE LIMITING ENZYMES 861 00:31:34,514 --> 00:31:37,217 ALSO KNOWN AS GSPDIN THE 862 00:31:37,217 --> 00:31:37,450 PATHWAY. 863 00:31:37,450 --> 00:31:38,251 AND INTERESTINGLY ENOUGH JUST A 864 00:31:38,251 --> 00:31:39,653 FEW YEARS BEFORE WE DID THIS 865 00:31:39,653 --> 00:31:42,389 WORK, WE HAVE DONE A WHOLE 866 00:31:42,389 --> 00:31:43,590 METABOLOMIC STUDY OF OUR FLY 867 00:31:43,590 --> 00:31:46,426 MODEL AND FOUND THAT BOTH TBLI 868 00:31:46,426 --> 00:31:52,098 COLSIS AND PHOSPHATE RELATED 869 00:31:52,098 --> 00:31:56,369 METABOLITES WERE ALTERED WITH 870 00:31:56,369 --> 00:31:57,037 PENTHOSPHOSPHATE WERE REDUCED 871 00:31:57,037 --> 00:31:58,471 CONSISTENT WITH THE FACT THAT WE 872 00:31:58,471 --> 00:32:00,440 MIGHT HAVE A DEFECT IN THE 873 00:32:00,440 --> 00:32:03,176 PATHWAY POTENTIALLY BECAUSE THE 874 00:32:03,176 --> 00:32:08,949 RATE LIMITING ENZYME GP D IS NOT 875 00:32:08,949 --> 00:32:11,151 TRANSLATED SUFFICIENTLY AS TDP43 876 00:32:11,151 --> 00:32:13,587 AS INDICATED BY RIBOTAGGING. 877 00:32:13,587 --> 00:32:16,289 AND THE METABOLOMICS DATA WERE 878 00:32:16,289 --> 00:32:20,994 CONFIRMED USING QPC R OF VARIOUS 879 00:32:20,994 --> 00:32:22,429 -- I'M JUST GOING TO SHOW YOU 880 00:32:22,429 --> 00:32:23,930 FROM HERE, IT'S HARD FOR ME TO 881 00:32:23,930 --> 00:32:27,200 SEE HERE BUT FROM HERE THIS IS 882 00:32:27,200 --> 00:32:28,835 G6 PC D IN CONTRAST TO THE 883 00:32:28,835 --> 00:32:30,036 MUTANT SYSTEM HIGHER IN CONTROLS 884 00:32:30,036 --> 00:32:31,771 AND IN PATIENT TISSUES, AS WELL 885 00:32:31,771 --> 00:32:36,509 AS IN PATIENT DERIVED NEURONS, 886 00:32:36,509 --> 00:32:39,379 THE TRANSCRIPT OF G6 PD IS 887 00:32:39,379 --> 00:32:40,814 INCREASED AND YOU CAN -- 888 00:32:40,814 --> 00:32:42,716 PROBABLY LOOKING HERE AT 889 00:32:42,716 --> 00:32:43,950 COMPENSATORY MECHANISM, WHERE 890 00:32:43,950 --> 00:32:46,186 YOU KNOW WE HYPOTHESIZED THAT 891 00:32:46,186 --> 00:32:47,988 THE TRANSLATION OF G6 PD'S 892 00:32:47,988 --> 00:32:51,024 REDUCED THERE IS PERHAPS A 893 00:32:51,024 --> 00:32:52,626 COMPENSATORY MECHANISM AT THE 894 00:32:52,626 --> 00:32:53,460 LEVEL OF TRANSCRIPTION TRYING TO 895 00:32:53,460 --> 00:32:55,128 MAKE MORE OF THIS TRANSCRIPT TO 896 00:32:55,128 --> 00:32:56,963 ULRICH MATILY MAKE MORE OF THIS 897 00:32:56,963 --> 00:32:57,530 PROTEIN. 898 00:32:57,530 --> 00:32:59,566 AND WE ADDRESSED THIS ISSUE 899 00:32:59,566 --> 00:33:02,235 USING -- WE ADDRESS THIS 900 00:33:02,235 --> 00:33:03,036 QUESTION USING GENETIC 901 00:33:03,036 --> 00:33:05,839 INTERACTION WHEREBY WE OVER 902 00:33:05,839 --> 00:33:07,173 EXPRESS GSPD AND USING THAT 903 00:33:07,173 --> 00:33:10,944 ASSAY I SHOWED YOU, YOU CAN SEE 904 00:33:10,944 --> 00:33:11,911 THAT TDPOVEREXPRESSION CAUSES A 905 00:33:11,911 --> 00:33:13,313 DEFECT, THERE WAS A HIGHER 906 00:33:13,313 --> 00:33:16,750 TURNING TIME, BUT THIS TIME, IS 907 00:33:16,750 --> 00:33:19,219 REDUCED FOR MUTANT TDP43 WHEN WE 908 00:33:19,219 --> 00:33:20,920 OVEREXPRESS G6 PD, SO THIS 909 00:33:20,920 --> 00:33:23,490 TURNING TIME IS SMALLER, 910 00:33:23,490 --> 00:33:25,125 COMPARED TO TDP43 ALONE. 911 00:33:25,125 --> 00:33:28,528 SUGGESTING THAT IF WE 912 00:33:28,528 --> 00:33:30,764 OVEREXPRESS G6 PD, POTENTIALLY 913 00:33:30,764 --> 00:33:36,469 RESTORE THE PENTOS-PHOSPHATE 914 00:33:36,469 --> 00:33:37,604 PATHWAY WE SEE THIS IN THE 915 00:33:37,604 --> 00:33:38,038 MODEL. 916 00:33:38,038 --> 00:33:40,807 IT WAS NICE THAT CAME TOGETHER 917 00:33:40,807 --> 00:33:46,179 WITH THE RIBOTAGGING APPROACH OR 918 00:33:46,179 --> 00:33:46,980 THE METABOLOMICS APPROACH. 919 00:33:46,980 --> 00:33:49,749 SO, THIS IS THE WORK WE'VE DONE 920 00:33:49,749 --> 00:33:52,986 TO MODEL ALS WHICH IS 921 00:33:52,986 --> 00:33:55,889 ESSENTIALLY TDP43 PROTEIN OP A 922 00:33:55,889 --> 00:33:57,557 THY IN MOTOR NEURONS BUT I 923 00:33:57,557 --> 00:33:59,659 HIGHLIGHTED TO YOU EARLIER THAT 924 00:33:59,659 --> 00:34:02,629 ALS AND FTD ARE A SPECTRUM 925 00:34:02,629 --> 00:34:05,165 DISORDER WHERE TDP43 926 00:34:05,165 --> 00:34:07,434 CHARACTERIZEs A LARGE, 927 00:34:07,434 --> 00:34:09,836 PROTEINS OP A THY CHARACTERIZES 928 00:34:09,836 --> 00:34:11,037 A LARGE FRACTION OF PATIENTS AND 929 00:34:11,037 --> 00:34:12,472 WE WANTED TO SEE WHETHER WE CAN 930 00:34:12,472 --> 00:34:14,040 FOCUS ON THE MUSHROOM BODY 931 00:34:14,040 --> 00:34:19,779 CIRCUIT IN THE FLY BRAIN, TO 932 00:34:19,779 --> 00:34:23,416 MODEL DEMENTIA AND A FORMER VERY 933 00:34:23,416 --> 00:34:26,386 TALENTED POST DOC IN MY LAB, 934 00:34:26,386 --> 00:34:27,520 OVEREXPRESSED TDP43 IN MUSHROOM 935 00:34:27,520 --> 00:34:28,588 NEURONS AND I HOPE YOU CAN SEE 936 00:34:28,588 --> 00:34:30,223 HERE THAT EARLY ON IN THE LARVA, 937 00:34:30,223 --> 00:34:33,460 MOST OF THE PROTEIN TAG WIDE YFP 938 00:34:33,460 --> 00:34:37,230 IS IN THE NUCLEUS, BUT IN ADULT 939 00:34:37,230 --> 00:34:39,099 FLIES, THE PROTEIN COMES OUT OF 940 00:34:39,099 --> 00:34:41,067 THE NUCLEUS AND YOU CAN SEE THIS 941 00:34:41,067 --> 00:34:42,335 HERE WITH THE DOUBLE ARROWS AND 942 00:34:42,335 --> 00:34:46,373 THE ARROW, CAN YOU SEE THESE 943 00:34:46,373 --> 00:34:47,707 MISLOCALLIZATION FROM THE 944 00:34:47,707 --> 00:34:49,142 NUCLEUS TO THE CYTOPLASM THAT 945 00:34:49,142 --> 00:34:50,610 IT'S OBSERVED IN PATIENTS. 946 00:34:50,610 --> 00:34:54,614 SHE THEN DID AN ANALYSIS OF 947 00:34:54,614 --> 00:34:56,716 NUCLEAR TO CYTOPLASMIC RATIO OF 948 00:34:56,716 --> 00:34:58,118 TDP43, SO WITHOUT GOING THROUGH 949 00:34:58,118 --> 00:34:59,219 ALL THE DETAILS OF THIS FIGURE, 950 00:34:59,219 --> 00:35:00,987 I WANT TO DRAW YOUR ATTENTION TO 951 00:35:00,987 --> 00:35:01,521 THE GRAPH. 952 00:35:01,521 --> 00:35:03,857 IN LARVA MOST OF THE PROTEIN IS 953 00:35:03,857 --> 00:35:07,293 IN THE NUCLEUS, IN YOUNG ADULTS 954 00:35:07,293 --> 00:35:09,262 ABOUT DAY 1-3, A LOT OF THE 955 00:35:09,262 --> 00:35:10,363 PROTEIN COMES OUT OF THE 956 00:35:10,363 --> 00:35:13,700 NUCLEUS, SO YOU SEE THIS REDUCED 957 00:35:13,700 --> 00:35:15,502 NUCLEUS TO CYTOPLASMIC RATIO, 958 00:35:15,502 --> 00:35:18,738 BUT TO OUR SURPRISES, ALL FLIES 959 00:35:18,738 --> 00:35:20,707 AROUND 50 OR SO DAYS OF AGE, 960 00:35:20,707 --> 00:35:24,477 THERE ARE MORE NEURONS WITH MORE 961 00:35:24,477 --> 00:35:26,813 TDP43 IN THE NUCLEUS THAN THE 962 00:35:26,813 --> 00:35:28,848 CYTOPLASM WHICH SEEMED COUNTER 963 00:35:28,848 --> 00:35:29,282 INTUITIVE. 964 00:35:29,282 --> 00:35:30,617 ON HOWEVER WHEN WE PAID 965 00:35:30,617 --> 00:35:31,818 ATTENTION AND STARTED COUNTING 966 00:35:31,818 --> 00:35:33,119 THE REMAINING NEURONS IN THE FLY 967 00:35:33,119 --> 00:35:34,854 BRAIN AT DAY 50, WE FOUND THAT 968 00:35:34,854 --> 00:35:37,357 YOU CAN SEE THIS IN THIS GRAPH, 969 00:35:37,357 --> 00:35:39,359 THERE WERE FEWER NEURONS ALIVE, 970 00:35:39,359 --> 00:35:41,594 IN THE OLD BRAIN, SUGGESTING 971 00:35:41,594 --> 00:35:44,464 THAT THE NEURONS THAT SURVIVED 972 00:35:44,464 --> 00:35:46,733 ARE ABLE TO RETAIN TDP 43 IN THE 973 00:35:46,733 --> 00:35:48,802 NUCLEUS, AND THEN WE CONFIRM 974 00:35:48,802 --> 00:35:50,203 THIS WITH KASP A R PACE 975 00:35:50,203 --> 00:35:52,205 STAININGS, THE CELLS THAT ARE 976 00:35:52,205 --> 00:35:55,341 LOSING TDP43 IN THE NUCLEUS, AND 977 00:35:55,341 --> 00:35:56,776 TDP43, MISLOCALLIZES TO THE 978 00:35:56,776 --> 00:35:59,312 CYTOPLASM, THOSE CELLS ARE LOST 979 00:35:59,312 --> 00:36:00,947 DURING LIFE SPAN AND HENCE THE 980 00:36:00,947 --> 00:36:03,383 REDUCTION IN OVERALL NEURONS 981 00:36:03,383 --> 00:36:04,484 WITH AGE. 982 00:36:04,484 --> 00:36:08,154 WITH THEN LOOKED AT A NUMBER OF 983 00:36:08,154 --> 00:36:08,922 PHENOTYPES INCLUDING AXONAL 984 00:36:08,922 --> 00:36:11,825 DEGENERATION, SO IF YOU LOOK IN 985 00:36:11,825 --> 00:36:12,625 TDP43 OVEREXPRESSING FLIES, YOU 986 00:36:12,625 --> 00:36:14,861 CAN SEE A THINNING OF THE AXONS, 987 00:36:14,861 --> 00:36:18,164 THIS IS THE CARTOON OF THE 988 00:36:18,164 --> 00:36:20,366 MUSHROOM BODY CIRCUIT, 989 00:36:20,366 --> 00:36:21,868 ALPHABETTA, GAMMA, NEURONS, AND 990 00:36:21,868 --> 00:36:23,069 YOU CAN SEE THEY ARE THINNING 991 00:36:23,069 --> 00:36:25,872 WITH AGE AND YOU CAN SEE THIS 992 00:36:25,872 --> 00:36:28,374 QUANTIFIED OVER HERE AND WE ALSO 993 00:36:28,374 --> 00:36:33,346 SEE AN INCREASE IN THE PUNTA 994 00:36:33,346 --> 00:36:35,181 THAT THE TDP43 FORMS AND YOU CAN 995 00:36:35,181 --> 00:36:36,483 SEE THIS QUANTIFIED OVER HERE, 996 00:36:36,483 --> 00:36:38,852 SO DURING LIFE SPAN THESE AXONS 997 00:36:38,852 --> 00:36:41,788 ARE DEGENERATING THE LOBES ARE 998 00:36:41,788 --> 00:36:46,359 THINNER AND THE TDP43 PANTA ARE 999 00:36:46,359 --> 00:36:47,894 ACCUMULATED AND INCREASED WITH 1000 00:36:47,894 --> 00:36:48,461 SIZE IN AGE. 1001 00:36:48,461 --> 00:36:50,230 WE THEN WANTED TO SEE IF WE CAN 1002 00:36:50,230 --> 00:36:51,397 MEASURE A BEHAVIORIAL ASSAY 1003 00:36:51,397 --> 00:36:58,972 THAT'S REALLY VANT TO DEMENTIA. 1004 00:36:58,972 --> 00:37:01,574 AND WE COLLABORATED WITH A CLOAG 1005 00:37:01,574 --> 00:37:04,544 AT ARIZONA WHO MADE THIS FOR 1006 00:37:04,544 --> 00:37:05,011 FLIES. 1007 00:37:05,011 --> 00:37:09,883 YOU MAY BE FAMILIAR WITH THIS 1008 00:37:09,883 --> 00:37:11,684 ASSAY FROM RODENTS, THEY'RE USED 1009 00:37:11,684 --> 00:37:13,253 TO QUANT TIE SPATIAL MEMORY AND 1010 00:37:13,253 --> 00:37:17,357 WHAT WE FOUND IS HERE'S A LITTLE 1011 00:37:17,357 --> 00:37:17,590 MOVIE. 1012 00:37:17,590 --> 00:37:20,860 WE PUT INDIVIDUAL FLIES IN THESE 1013 00:37:20,860 --> 00:37:23,196 LITTLE Y-MAZE AND WE IMAGE THEM 1014 00:37:23,196 --> 00:37:24,964 FOR 10 MINUTES THEN WE EVALUATE 1015 00:37:24,964 --> 00:37:26,900 THE MOVEMENT AND THE ABILITY TO 1016 00:37:26,900 --> 00:37:30,570 SHIFT ARMS IN IN PERIOD OF TIME. 1017 00:37:30,570 --> 00:37:34,474 AND WE FOUND THAT WHEN WE 1018 00:37:34,474 --> 00:37:37,310 MEASURED DISTANCE, AND THE 1019 00:37:37,310 --> 00:37:39,979 ALTASDINATION MEANING ABILITY TO 1020 00:37:39,979 --> 00:37:41,214 CHANGE ARMS CONSECUTIVELY, WE 1021 00:37:41,214 --> 00:37:43,383 FIND THAT ALTHOUGH THE OVERALL 1022 00:37:43,383 --> 00:37:44,784 MOVEMENT EXPRESSION IN MUSHROOM 1023 00:37:44,784 --> 00:37:47,520 BODY WAS HIGHER, THE 1024 00:37:47,520 --> 00:37:48,087 ALTASDINATION WAS REDUCED 1025 00:37:48,087 --> 00:37:50,123 CONSIST EPT WITH THE DEFECT IN 1026 00:37:50,123 --> 00:37:51,791 SPATIAL WORKING MEMORY IN THESE 1027 00:37:51,791 --> 00:37:52,559 FLIES. 1028 00:37:52,559 --> 00:37:55,595 AND THAT WAS THROUGH BOTH FOR 1029 00:37:55,595 --> 00:37:58,364 MALES AND FEMALES. 1030 00:37:58,364 --> 00:38:00,667 WE THEN LOOKED AT WHETHER THERE 1031 00:38:00,667 --> 00:38:06,072 ARE DEFECT IN SLEEP, AND JUST TO 1032 00:38:06,072 --> 00:38:08,675 FOCUS ON THE KEY POINT HERE IS 1033 00:38:08,675 --> 00:38:11,210 THAT TO OUR SURPRISE, WE FOUND 1034 00:38:11,210 --> 00:38:15,415 INCREASED SLEEP CAUSED BY TDP43. 1035 00:38:15,415 --> 00:38:17,350 WHICH IS KIND OF UNEXPECTED BUT 1036 00:38:17,350 --> 00:38:19,152 THEN AS I READ MORE ABOUT 1037 00:38:19,152 --> 00:38:21,621 PATIENT PHENOTYPES, IT TURNS OUT 1038 00:38:21,621 --> 00:38:23,022 PATIENTS WITH DEMENTIA DO SLEEP 1039 00:38:23,022 --> 00:38:25,758 MORE DURING THE DAY BUT THEY DO 1040 00:38:25,758 --> 00:38:29,295 EXHIBIT SLEEP FRAGMENTATION 1041 00:38:29,295 --> 00:38:29,796 DURING THE NIGHT. 1042 00:38:29,796 --> 00:38:33,166 AND THE ONLY TIME OR THE ONLY 1043 00:38:33,166 --> 00:38:36,469 PLACE WHERE WE IDENTIFIED SLEEP 1044 00:38:36,469 --> 00:38:37,837 FRAGMENTATION OF TDP43 1045 00:38:37,837 --> 00:38:39,238 OVEREXPRESSION IN BODIES IS IN 1046 00:38:39,238 --> 00:38:40,740 YOUNG MALES BUT YOU CAN SEE THAT 1047 00:38:40,740 --> 00:38:42,775 AT NIGHT THERE ARE MORE SLEEP 1048 00:38:42,775 --> 00:38:43,910 BOUTS BUT THERE ARE SHORTER. 1049 00:38:43,910 --> 00:38:46,846 SO THE SLEEP IS MORE FRAGMENTED 1050 00:38:46,846 --> 00:38:47,380 AT NIGHT. 1051 00:38:47,380 --> 00:38:49,082 LATTER ON THE PHENOTYPES BECOME 1052 00:38:49,082 --> 00:38:50,583 MORE COMPLEX, BUT AT LEAST THIS 1053 00:38:50,583 --> 00:38:53,920 1 POINT IN TIME, AT LEAST FOR 1054 00:38:53,920 --> 00:38:55,021 MAN'S EXPRESSING TDP43 AND 1055 00:38:55,021 --> 00:38:57,357 KENYAN CELLS WE COULD IDENTIFY 1056 00:38:57,357 --> 00:38:59,225 SLEEP FRAGMENTATION, SIMILAR TO 1057 00:38:59,225 --> 00:38:59,926 WHAT'S IN PATIENTS. 1058 00:38:59,926 --> 00:39:01,661 THERE IS ALSO A SLIGHT BUT 1059 00:39:01,661 --> 00:39:02,929 SIGNIFICANT REDUCTION IN LIFE 1060 00:39:02,929 --> 00:39:04,731 SPAN, WHICH WAS KIND OF 1061 00:39:04,731 --> 00:39:06,666 UNEXPECTED BUT ALSO INTERESTING 1062 00:39:06,666 --> 00:39:09,002 BECAUSE WE'RE ONLY MANIPULATING 1063 00:39:09,002 --> 00:39:09,969 KENYAN CELLS HERE, NOT ALL THE 1064 00:39:09,969 --> 00:39:13,106 NEURONS IN THE BRAIN. 1065 00:39:13,106 --> 00:39:18,878 AND WE ALSO USED A NONCANNONICLE 1066 00:39:18,878 --> 00:39:21,447 ACID APPROACH TO SEE WHEN TDP43 1067 00:39:21,447 --> 00:39:22,749 IN THE KENYAN CELLS CAUSES A 1068 00:39:22,749 --> 00:39:24,150 REDUCTION IN THE NEW PROTEIN 1069 00:39:24,150 --> 00:39:25,318 SYNTHESIS AND THIS IS THE FUN 1070 00:39:25,318 --> 00:39:26,386 CAT VERSION OF THIS EXPERIMENT 1071 00:39:26,386 --> 00:39:28,054 AND YOU CAN SEE THAT THAT THERE 1072 00:39:28,054 --> 00:39:30,490 IS A REDUCTION IN SIGNAL IN THE 1073 00:39:30,490 --> 00:39:32,625 CONTEXT OF TDP43, SO WE'RE 1074 00:39:32,625 --> 00:39:34,794 CURRENTLY USING THE BONCAT 1075 00:39:34,794 --> 00:39:39,499 APPROACH TO IDENTIFY WHAT ARE 1076 00:39:39,499 --> 00:39:43,770 NOT DONE CORRECTLY IN THIS 1077 00:39:43,770 --> 00:39:44,437 CIRCUIT. 1078 00:39:44,437 --> 00:39:45,772 SO THE REASON -- 1 OF THE 1079 00:39:45,772 --> 00:39:48,041 REASONS WE WANTED TO MANIPULATE 1080 00:39:48,041 --> 00:39:50,910 THE CIRC YOUS TO DEVELOP ALS 1081 00:39:50,910 --> 00:39:52,445 RELEVANT MODEL BECAUSE OF 1082 00:39:52,445 --> 00:39:53,012 DEMENTIA RELEVANT MODEL IS 1083 00:39:53,012 --> 00:39:55,148 BECAUSE WE WANT TO LOOK AT THE 1084 00:39:55,148 --> 00:39:56,549 OVERLAP AS WELL AS UNIQUE 1085 00:39:56,549 --> 00:39:59,218 FEATURES THAT MAY BE RELEVANT TO 1086 00:39:59,218 --> 00:40:00,753 ALS OR FTD SPECIFICALLY AND WE 1087 00:40:00,753 --> 00:40:02,522 DID AN OVERLAP OF THE MRNAs 1088 00:40:02,522 --> 00:40:04,290 THAT ARE ASSOCIATED WITH TDP43, 1089 00:40:04,290 --> 00:40:06,159 WANT TO DO THIS EXPERIMENT IN 1090 00:40:06,159 --> 00:40:08,461 MOTOR NEURONS VERSUS KENYAN 1091 00:40:08,461 --> 00:40:11,964 CELLS, IN THE DEMENTIA MODEL, 1092 00:40:11,964 --> 00:40:14,200 AND THE REASON I SHOWED YOU DLP 1093 00:40:14,200 --> 00:40:16,335 BEFORE IS BECAUSE WE ALSO FOUND 1094 00:40:16,335 --> 00:40:18,805 DLP AS A POTENTIAL TARGET OF 1095 00:40:18,805 --> 00:40:20,206 TDP43 IN THE MUSHROOM BODIES. 1096 00:40:20,206 --> 00:40:22,108 SO THIS MAY BE 1 OF THE THOSE 1097 00:40:22,108 --> 00:40:24,444 MRNAs THAT IS REGULATED BY 1098 00:40:24,444 --> 00:40:26,779 TDP43, BOTH IN ALS, AS WELL AS 1099 00:40:26,779 --> 00:40:29,649 IN THE CONTEXT OF FTD, WHILE 1100 00:40:29,649 --> 00:40:32,251 OTHER MRNARKs ARE SPECIFICALLY 1101 00:40:32,251 --> 00:40:33,853 REGULATED BY TDP43 MOTOR NEURONS 1102 00:40:33,853 --> 00:40:35,822 OR SPECIFICALLY IN THE MUSHROOM 1103 00:40:35,822 --> 00:40:37,256 BODY CIRCUIT, AND WE HOPE THAT 1104 00:40:37,256 --> 00:40:41,394 THIS APPROACH IS GIVING US THE 1105 00:40:41,394 --> 00:40:43,730 ABILITY TO IDENTIFY THE 1106 00:40:43,730 --> 00:40:44,530 COMMONALITIES BETWEEN THESE 2 1107 00:40:44,530 --> 00:40:47,366 FORMS OF THE SPECTRUM DISORDER, 1108 00:40:47,366 --> 00:40:49,669 AS WELL AS SPECIFIC 1109 00:40:49,669 --> 00:40:52,171 MANIFESTATIONS OF THESE 2. 1110 00:40:52,171 --> 00:41:00,513 SO, THIS IS DLP EXPRESSION IN 1111 00:41:00,513 --> 00:41:01,681 YOUNG VERSUS OLD FLIES AND IT'S 1112 00:41:01,681 --> 00:41:03,750 A LITTLE BIT DIFFICULT BECAUSE 1113 00:41:03,750 --> 00:41:05,818 OF AGE DLP KIND OF EXPRESSES 1114 00:41:05,818 --> 00:41:07,386 EVERYWHERE BUT WE CAN HIGHLIGHT 1115 00:41:07,386 --> 00:41:09,088 THE MUSHROOM BODY SPECIFICALLY 1116 00:41:09,088 --> 00:41:10,123 CAN DIFFERENT MARKIERS AND WHAT 1117 00:41:10,123 --> 00:41:13,259 WE FOUND IS THAT ALTHOUGH DURING 1118 00:41:13,259 --> 00:41:15,461 AGING DLP IN A NORMAL CONDITION 1119 00:41:15,461 --> 00:41:17,096 IS REDUCED IN EXPRESSION IS 1120 00:41:17,096 --> 00:41:20,199 REDUCED EVEN FURTHER BY TDP 43 1121 00:41:20,199 --> 00:41:22,301 OVEREXPRESSION, AND JUST LIKE IN 1122 00:41:22,301 --> 00:41:26,272 MOTOR NEURONS IN THE MUSHROOM 1123 00:41:26,272 --> 00:41:27,173 BODY CIRCUIT, DLP IS THOUGHT TO 1124 00:41:27,173 --> 00:41:29,375 PLAY A REEL KEEPSAKES IN 1125 00:41:29,375 --> 00:41:31,911 SIGNALING AS WELL AS SYNAPTIC 1126 00:41:31,911 --> 00:41:32,211 PLASTICITY. 1127 00:41:32,211 --> 00:41:35,348 AND THEN WE WANTED TO KNOW -- WE 1128 00:41:35,348 --> 00:41:37,683 TOOK ADVANTAGE OF POWER OF FLY 1129 00:41:37,683 --> 00:41:40,419 GENETICS AND WE WANTED TO KNOW 1130 00:41:40,419 --> 00:41:43,189 WHETHER OVEREXPRESSION OF DLP 1131 00:41:43,189 --> 00:41:44,824 CAN RESCUE TDP 43 DEPENDENT 1132 00:41:44,824 --> 00:41:46,492 PHENOTYPE, SO USING THAT ASSAY I 1133 00:41:46,492 --> 00:41:49,662 SHOWED YOU, YOU CAN SEE THAT DLP 1134 00:41:49,662 --> 00:41:50,930 OVEREXPRESSION RESCUES THE 1135 00:41:50,930 --> 00:41:53,332 MOVEMENT DEFICIT AND IT ALSO 1136 00:41:53,332 --> 00:41:56,602 RESCUES THE ALTASDINATION 1137 00:41:56,602 --> 00:41:57,069 DEFICIT. 1138 00:41:57,069 --> 00:42:00,540 AND THEN WE WERE ASKING 1139 00:42:00,540 --> 00:42:01,707 OURSELVES, OKAY, THIS WORKS 1140 00:42:01,707 --> 00:42:06,846 GREAT IN FLIES BUT HOW ABOUT IN 1141 00:42:06,846 --> 00:42:09,348 PATIENTS AND WE TRIED OUR HANDS 1142 00:42:09,348 --> 00:42:13,386 AT STAINING FOR THE ORTHOLOG 1143 00:42:13,386 --> 00:42:14,887 GP16 FTD PATIENT TISSUES BUT THE 1144 00:42:14,887 --> 00:42:16,622 STAINING IS VERY COMPLEX AND 1145 00:42:16,622 --> 00:42:17,557 HETEROGENEOUS ORY GENIUS, SO 1146 00:42:17,557 --> 00:42:19,091 THEN WE TOOK ADVABTAGE OF A DATA 1147 00:42:19,091 --> 00:42:21,961 SET THAT WAS GENERATED BY OUR 1148 00:42:21,961 --> 00:42:24,463 COLLABORATORS WHERE THEY DID 1149 00:42:24,463 --> 00:42:26,365 SINGLE NUCLEUS RNA SEQ, FROM FTD 1150 00:42:26,365 --> 00:42:28,134 PATIENT BRAINS AND WHAT THEY DID 1151 00:42:28,134 --> 00:42:31,604 IS THEY LOOKED WITHIN THE FTD 1152 00:42:31,604 --> 00:42:33,606 PATIENT BRAIN POPULATION AT 1153 00:42:33,606 --> 00:42:35,708 NEURONS THAT STILL HAVE TDP43 IN 1154 00:42:35,708 --> 00:42:39,245 THE NUCLEUS, VERSUS NEURONS IN 1155 00:42:39,245 --> 00:42:41,414 THAT PATIENT POPULATION THAT 1156 00:42:41,414 --> 00:42:43,015 LOST TDP43 IN THE NUCLEUS, 1157 00:42:43,015 --> 00:42:44,150 MEANING THEY HAVE THE PATHOLOGY 1158 00:42:44,150 --> 00:42:45,918 AND THIS CAN BE DETERMINED VIA 1159 00:42:45,918 --> 00:42:48,454 THE PRESENCE OF CRYPTICAXONS, 1160 00:42:48,454 --> 00:42:50,590 THAT ARE ASSOCIATED WITH LOSS OF 1161 00:42:50,590 --> 00:42:53,226 TDP43 FROM THE NUCLEUS, SO WHEN 1162 00:42:53,226 --> 00:42:56,329 DOING THIS KIND OF ANALYSIS, 1163 00:42:56,329 --> 00:42:58,231 DIFFERENTIAL GENE EXPRESSION 1164 00:42:58,231 --> 00:42:59,632 WITHIN THE FTD POPULATION, 1165 00:42:59,632 --> 00:43:01,200 BETWEEN NEURONS THAT HAVE NORMAL 1166 00:43:01,200 --> 00:43:05,504 TDP 43 IN THE NUCLEUS, VERSUS 1167 00:43:05,504 --> 00:43:06,405 TDP 43 PATHOLOGY, GUESS WHICH 1168 00:43:06,405 --> 00:43:10,376 WAS 1 OF THE BEST-WHICH WAS 1 OF 1169 00:43:10,376 --> 00:43:13,279 THE TARGETS, 1 OF THE MOST 1170 00:43:13,279 --> 00:43:15,681 DIFFERENTIAL EXPRESSED GENES WAS 1171 00:43:15,681 --> 00:43:18,851 GPC 6, WHICH WAS OUR DLP 1172 00:43:18,851 --> 00:43:19,218 ORTHOLOG. 1173 00:43:19,218 --> 00:43:21,120 SO ESSENTIALLY THIS IS TELLING 1174 00:43:21,120 --> 00:43:22,622 US THAT ALTHOUGH IT'S NOT 1175 00:43:22,622 --> 00:43:23,990 NECESSARILY THE SAME EXPERIMENT, 1176 00:43:23,990 --> 00:43:25,658 NOT THE SAME DIRECTION THAT IT 1177 00:43:25,658 --> 00:43:30,863 HAPPENS IN THE FLY, WE CAN LINK 1178 00:43:30,863 --> 00:43:32,531 DLP/GPC 6 TO TDP43 PATHOLOGY 1179 00:43:32,531 --> 00:43:34,901 FROM FLY ALL THE WAY TO PATIENT 1180 00:43:34,901 --> 00:43:35,735 TISSUES. 1181 00:43:35,735 --> 00:43:38,004 SO TO SUMMARIZE THIS FESTER PART 1182 00:43:38,004 --> 00:43:42,408 OF MY TALK, WHICH TALK TOOK MORE 1183 00:43:42,408 --> 00:43:44,810 THAN A THOUGHT, I WANTED TO 1184 00:43:44,810 --> 00:43:47,947 HIGHLIGHT TO YOU THE POWER OF 1185 00:43:47,947 --> 00:43:48,981 MODELING TDPPROTEIN OPERATING 1186 00:43:48,981 --> 00:43:50,716 GLOBALLYATHY IN THE CIRCUITS 1 1187 00:43:50,716 --> 00:43:53,486 THAT'S RELEVANT TO ALS AND 1 1188 00:43:53,486 --> 00:43:55,321 THAT'S RELEVANT TO FTD AND THE 1189 00:43:55,321 --> 00:43:56,722 DISCOVERIES WE MADE IN THE FLY 1190 00:43:56,722 --> 00:43:59,125 THAT MORE THAN 90% EVEN TO MY 1191 00:43:59,125 --> 00:44:01,794 SURPRISE AS A FLY GENETICIST 1192 00:44:01,794 --> 00:44:03,829 VALIDATED PATIENT TISSUES ALMOST 1193 00:44:03,829 --> 00:44:04,931 EVERY SINGLE TIME, WERE TARGETS 1194 00:44:04,931 --> 00:44:08,434 THAT HAVE TO DO WITH 1195 00:44:08,434 --> 00:44:10,136 CYTOSKELETON FUNCTION, MICROTUBE 1196 00:44:10,136 --> 00:44:13,339 Y'ALL ABILITY, SYNAPTIC VESICLE 1197 00:44:13,339 --> 00:44:15,541 CYCLE, METABOLISM AND WIN 1198 00:44:15,541 --> 00:44:18,311 SIGNALING, WITH -- WE'RE PROUD 1199 00:44:18,311 --> 00:44:19,612 OF THIS FTD MODEL BECAUSE IT 1200 00:44:19,612 --> 00:44:21,113 ALLOWS US TO DO ADDITIONAL 1201 00:44:21,113 --> 00:44:21,647 GENETIC SCREENS THAT ARE 1202 00:44:21,647 --> 00:44:24,617 SPECIFIC TO THE CIRCUIT AS WELL 1203 00:44:24,617 --> 00:44:26,986 AS DRUG SCREENS, AND DLPs, THE 1204 00:44:26,986 --> 00:44:29,422 FIRST TARGET WE FOCUSED ON, 1205 00:44:29,422 --> 00:44:30,656 BECAUSE IT ALLOWED US TO TIE 1206 00:44:30,656 --> 00:44:32,458 INTO THE 2 MODELS BUT THERE ARE 1207 00:44:32,458 --> 00:44:33,859 MANY, MANY MORE INTERESTING 1208 00:44:33,859 --> 00:44:36,462 TARGETS THAT ARE ALS SPECIFIC 1209 00:44:36,462 --> 00:44:37,396 VERSUS FTD SPECIFIC THATY WOO 1210 00:44:37,396 --> 00:44:41,867 WOULD LIKE -- THAT WE WOULD LIO 1211 00:44:41,867 --> 00:44:42,935 WORK ON. 1212 00:44:42,935 --> 00:44:45,404 WE SO THIS MISLOCALLIZATION IN 1213 00:44:45,404 --> 00:44:47,306 THE CELL DEATH OF THE FTD MODEL 1214 00:44:47,306 --> 00:44:50,242 AND WE WERE ABLE TO TIE IN THE 1215 00:44:50,242 --> 00:44:52,545 TARGETS THAT ARE DISREGULATED IN 1216 00:44:52,545 --> 00:44:55,414 THE FLY MODEL TO DATA FROM 1217 00:44:55,414 --> 00:44:57,049 PATIENT TISSUES WHERE THE SAME 1218 00:44:57,049 --> 00:45:00,720 TARGETS ARE ALSO ALTERED IN 1219 00:45:00,720 --> 00:45:02,121 NEURONS OF TDP43 PATHOLOGY AND 1220 00:45:02,121 --> 00:45:05,391 WHILE THESE MODELS OF 1221 00:45:05,391 --> 00:45:06,659 OVEREXPRESSION HAVE SIGNIFICANT 1222 00:45:06,659 --> 00:45:08,427 CAVEATS BECAUSE WE RECOGNIZE 1223 00:45:08,427 --> 00:45:10,730 THAT EVERY TIME YOU OVEREXPRESS 1224 00:45:10,730 --> 00:45:12,031 AN RNA BINDING PROTEIN THERE 1225 00:45:12,031 --> 00:45:13,899 WILL BE SOMEART FACTS, ON 1 HAND 1226 00:45:13,899 --> 00:45:15,501 IT WAS SATISFYING THAT WE WERE 1227 00:45:15,501 --> 00:45:17,370 ABLE TO VALIDATE IN-PATIENT 1228 00:45:17,370 --> 00:45:18,771 TISSUES, THE TARGETS WE FOUND IN 1229 00:45:18,771 --> 00:45:21,207 THE FLY, BUT AT THE SIEM TIME 1230 00:45:21,207 --> 00:45:22,608 WE'RE COGNIZANT THAT WE NEED TO 1231 00:45:22,608 --> 00:45:23,442 ALSO TAKE DIFFERENT APPROACHES 1232 00:45:23,442 --> 00:45:32,318 SO WE'VE BEEN USING A C RISPR 1233 00:45:32,318 --> 00:45:36,022 KNOCK-IN MODEL THAT REPLACES 1234 00:45:36,022 --> 00:45:38,824 TDPIN THE FLY WITH HUMAN TDP43, 1235 00:45:38,824 --> 00:45:40,092 WHETHER IT'S WILD-TYPE OR MUTANT 1236 00:45:40,092 --> 00:45:42,028 AND A VERY TALENTED STUDENT IN 1237 00:45:42,028 --> 00:45:44,497 MY LAB WANTS ON DO SINGLE CELL 1238 00:45:44,497 --> 00:45:48,768 RNA SEQ AND LOOKA THE EFFECTS OF 1239 00:45:48,768 --> 00:45:51,737 TDP43 PROTEIN OP A THY, DURING 1240 00:45:51,737 --> 00:45:51,937 AGING. 1241 00:45:51,937 --> 00:45:53,706 AND SOME OF YOU IN THIS ROOM 1242 00:45:53,706 --> 00:45:54,940 HELPED US DO THESE EXPERIMENTS, 1243 00:45:54,940 --> 00:45:56,909 SO THANK YOU VERY MUCH FOR THAT. 1244 00:45:56,909 --> 00:46:00,446 AND THIS IS THE LITERALLY DATA 1245 00:46:00,446 --> 00:46:01,781 AS OF MONDAY IT WAS REALLY 1246 00:46:01,781 --> 00:46:03,149 EXCITING TO SEE THAT OUR VERY 1247 00:46:03,149 --> 00:46:07,920 FIRST ATTEMPT WITH A LOT OF HELP 1248 00:46:07,920 --> 00:46:11,257 FROM ELLA'S LAB, ROSARIO IN 1249 00:46:11,257 --> 00:46:12,691 PARTICULAR, WE WERE ABLE TO 1250 00:46:12,691 --> 00:46:15,261 GENERATE DATA THAT LOOKED LIKE 1251 00:46:15,261 --> 00:46:16,295 IT REALLY ISOLATED BEAUTIFUL 1252 00:46:16,295 --> 00:46:17,329 CLUSTERS IN THE FLY BRAIN AND 1253 00:46:17,329 --> 00:46:20,900 FOR THIS 1 WE WANT ON DO A QUICK 1254 00:46:20,900 --> 00:46:21,967 DIFFERENTIAL GENE EXPRESSION IN 1255 00:46:21,967 --> 00:46:24,203 THE KENYAN CELLS WHICH REGULATE 1256 00:46:24,203 --> 00:46:25,504 SPATIAL MEMORY, SLEEP AND OTHER 1257 00:46:25,504 --> 00:46:27,673 BEHAVIORS THAT ARE RELEVANT TO 1258 00:46:27,673 --> 00:46:30,042 DISEASE, AND INTERESTINGLY 1259 00:46:30,042 --> 00:46:31,777 ENOUGH, WE FOUND SOME 1260 00:46:31,777 --> 00:46:32,945 DISREGULATED GENES SOME OF WHICH 1261 00:46:32,945 --> 00:46:34,346 ARE LINKED TO LEARNING AND 1262 00:46:34,346 --> 00:46:36,282 MEMORY SUCH AS DANCE SHOWN HERE. 1263 00:46:36,282 --> 00:46:37,917 SO THIS IS VERY NEW, WE'RE JUST 1264 00:46:37,917 --> 00:46:39,285 BEGINNING TO UNDERSTAND THAT, 1265 00:46:39,285 --> 00:46:41,253 BUT JUST TO GIVE YOU AN IDEA 1266 00:46:41,253 --> 00:46:42,455 THAT WHILE THE OVEREXPRESSION 1267 00:46:42,455 --> 00:46:45,057 MODELS HAVE TAUGHT US A LOT, 1268 00:46:45,057 --> 00:46:46,125 THAT VALIDATED IN PATIENT 1269 00:46:46,125 --> 00:46:48,094 TISSUES, WE ARE ALSO USING OTHER 1270 00:46:48,094 --> 00:46:51,730 APPROACHES SUCH AS THIS CRISPR 1271 00:46:51,730 --> 00:46:55,601 KNOCK INS TO UNDERSTAND TDP43 1272 00:46:55,601 --> 00:46:57,203 MECHANISMS, AND SO SINCE I ONLY 1273 00:46:57,203 --> 00:46:59,338 HAVE 13 MINUTES I HAVE 2 OTHER 1274 00:46:59,338 --> 00:47:06,979 STORIES I WANTED TO TALK ABOUT, 1275 00:47:06,979 --> 00:47:09,181 1 IS IN MUSCLES AND THE OTHER IS 1276 00:47:09,181 --> 00:47:12,451 METABOLISM SO I DON'T IF I 1277 00:47:12,451 --> 00:47:16,255 SHOULD TAKE A VOTE? 1278 00:47:16,255 --> 00:47:17,056 GO FOR IT. 1279 00:47:17,056 --> 00:47:18,724 >> METABOLISM BUT THAT'S THE 1280 00:47:18,724 --> 00:47:19,992 SECOND 1. 1281 00:47:19,992 --> 00:47:20,159 SO. 1282 00:47:20,159 --> 00:47:22,194 HOW MANY PEEP WANT TO SEE 1283 00:47:22,194 --> 00:47:23,596 METRICS TAB -- HOW MANY PEOPLE 1284 00:47:23,596 --> 00:47:24,396 WANT TO DO METABOLISM. 1285 00:47:24,396 --> 00:47:27,900 I WILL DO THIS, THIS IS SO COOL, 1286 00:47:27,900 --> 00:47:29,468 THE MUSCLE BON, NCAT DATA IS 1287 00:47:29,468 --> 00:47:31,770 AMAZING, SO LOOK AT THIS, YOU 1288 00:47:31,770 --> 00:47:32,571 OVEREXPRESS TDP43 IN THE 1289 00:47:32,571 --> 00:47:34,807 MUSCLES, THIS IS THE NEWEL 1290 00:47:34,807 --> 00:47:35,875 SYNTHESIZED PROTEIN IN THE 1291 00:47:35,875 --> 00:47:36,842 NORMAL MUSCLE, THIS IS REDUCTION 1292 00:47:36,842 --> 00:47:39,011 IN SIGNATURES WILL NAING CONTEXT 1293 00:47:39,011 --> 00:47:40,746 OF TDP43, AND THIS IS THE 1294 00:47:40,746 --> 00:47:43,415 VOLCANO PLOT OF THE BIOATTENUATE 1295 00:47:43,415 --> 00:47:44,383 PROTEINS SO THERE'S A LOT OF 1296 00:47:44,383 --> 00:47:47,820 THINGS THAT ARE ALTERED AND MANY 1297 00:47:47,820 --> 00:47:50,556 OF THEM, ARE LISTENED TO 1298 00:47:50,556 --> 00:47:52,625 METABOLISM, WHETHER IT'S TCA 1299 00:47:52,625 --> 00:47:56,262 CYCLE OR LIPID METABOLISM, 1300 00:47:56,262 --> 00:47:57,530 OROXIDATIVE STRESS, AND THIS IS 1301 00:47:57,530 --> 00:48:00,566 SOMETHING THAT WE'RE JUST NOW 1302 00:48:00,566 --> 00:48:03,169 BEGINNING TO LOOK AT AND 1303 00:48:03,169 --> 00:48:03,469 VALIDATE. 1304 00:48:03,469 --> 00:48:07,173 THERE ARE A NUMBER OF PHENOTYPES 1305 00:48:07,173 --> 00:48:09,909 THAT WE SEE IN THE MUSCLE. 1306 00:48:09,909 --> 00:48:12,111 AND 1 OF THE THINGS THAT WE SEE 1307 00:48:12,111 --> 00:48:16,248 IS A REDUCTION IN FRAGILE X 1308 00:48:16,248 --> 00:48:16,949 MENTAL RETARDATION PROTEIN, I 1309 00:48:16,949 --> 00:48:18,217 WAS TELLING SOMEONE THAT I 1310 00:48:18,217 --> 00:48:20,186 STARTED MY CAREER IN FRAGILE X 1311 00:48:20,186 --> 00:48:23,022 SYNDROME AND I MOVED TO AGE 1312 00:48:23,022 --> 00:48:23,689 DEPENDENT NEUROLOGICAL DISORDERS 1313 00:48:23,689 --> 00:48:25,024 ONLY TO FIND A CONNECTION 1314 00:48:25,024 --> 00:48:28,060 BETWEEN THE RNA BINDING PROTEINS 1315 00:48:28,060 --> 00:48:31,864 THAT PLAY A ROLE IN THE AGE OF 1316 00:48:31,864 --> 00:48:32,765 NEURODEGENERATION RESPECTIVELY 1317 00:48:32,765 --> 00:48:40,139 SO WE FOUND THAT FMRP IS 1318 00:48:40,139 --> 00:48:41,240 OVEREXPRESSED IN DDP43, SO AND 1319 00:48:41,240 --> 00:48:43,943 IF WE OVEREXPRESS THEM, WE SEE 1320 00:48:43,943 --> 00:48:51,350 THIS, AND RESCUE THE LEATHALLITY 1321 00:48:51,350 --> 00:48:52,384 IN TDP43 MUSCLE. 1322 00:48:52,384 --> 00:48:55,988 WE SEE STRUCTURAL DEFECTS AND IN 1323 00:48:55,988 --> 00:48:57,523 COLLABORATION WITH DR. CRUZ'S 1324 00:48:57,523 --> 00:49:02,428 LAB, WE FOUND IN THE MASS MODEL 1325 00:49:02,428 --> 00:49:08,133 OF TDP43 PROTEIN OP A THY, THE 1326 00:49:08,133 --> 00:49:09,235 ORTHOLOG, CALLED FXR1 IS REDUCED 1327 00:49:09,235 --> 00:49:10,536 IN THE MOUSE, SO THERE'S THE 1328 00:49:10,536 --> 00:49:13,372 FLY, THE MOUSE AND ALSO THE 1329 00:49:13,372 --> 00:49:17,142 HUMAN SO LOOKING AT MUSCLE, 1330 00:49:17,142 --> 00:49:19,545 POSTMORTEM SAMPLES FROM ALS 1331 00:49:19,545 --> 00:49:23,382 PATIENTS, WE CAN SEE WITH THIS 1332 00:49:23,382 --> 00:49:26,652 EXCEPTION HERE, A SPORADIC CASES 1333 00:49:26,652 --> 00:49:28,287 THAT HAVE TDP43 PROTEIN 1334 00:49:28,287 --> 00:49:30,422 OPERATING GLOBALLYATHY, THE 1335 00:49:30,422 --> 00:49:33,425 MUSCLE ALSO OF FMRP, IS REDUCED 1336 00:49:33,425 --> 00:49:35,861 IN PATIENT SAMPLES, HIGHLIGHTING 1337 00:49:35,861 --> 00:49:37,529 THE FLY TOA IDENTIFY TARGETS 1338 00:49:37,529 --> 00:49:38,597 THAT ARE REFINED WITH THE 1339 00:49:38,597 --> 00:49:40,566 PATIENT TISSUES AND USING 1340 00:49:40,566 --> 00:49:43,402 GENETIC APPROACHES WE'VE SHOWN 1341 00:49:43,402 --> 00:49:45,204 THAT RESTORING FMRP FUNCTION WE 1342 00:49:45,204 --> 00:49:48,207 CAN RESCUE THE TDP 43 ASSOCIATED 1343 00:49:48,207 --> 00:49:50,009 PHENOTYPES SO NOWIME READY TO GO 1344 00:49:50,009 --> 00:49:51,143 TO METABOLISM, LET'S SEE HOW 1345 00:49:51,143 --> 00:50:00,152 FAST CAN I SPEAK? 1346 00:50:00,152 --> 00:50:00,986 REALLY FAST. 1347 00:50:00,986 --> 00:50:02,721 SO THIS PROJECT CAME ABOUT BY 1348 00:50:02,721 --> 00:50:03,088 ACCIDENT. 1349 00:50:03,088 --> 00:50:05,391 ON 1 HAND, MY YOUNGER DAUGHTER 1350 00:50:05,391 --> 00:50:08,360 WHO WAS A TEENAGER AT THE TIME 1351 00:50:08,360 --> 00:50:09,995 WAS OBSESS WIDE METABOLISM AND 1352 00:50:09,995 --> 00:50:11,330 WHAT YOU SHOULD EAT TO MAINTAIN 1353 00:50:11,330 --> 00:50:13,399 YOUR FIGURE AND THEN I HAD A 1354 00:50:13,399 --> 00:50:14,366 GRADUATE STUDENT IN THE LAB WHO 1355 00:50:14,366 --> 00:50:15,768 WAS INTERESTED IN THE EEIVETS OF 1356 00:50:15,768 --> 00:50:18,937 FOOD AND DIFFERENT COMPONENTS OF 1357 00:50:18,937 --> 00:50:23,609 THE FOOD ON TDP43 PHENOTYPES SO 1358 00:50:23,609 --> 00:50:24,777 THIS PROMPTED STUDYING THE 1359 00:50:24,777 --> 00:50:28,180 METRICS TAB O LOAM OF ANIMALS 1360 00:50:28,180 --> 00:50:29,948 WITH TDP43 PROTEIN OP A THY, BUT 1361 00:50:29,948 --> 00:50:32,918 IT WAS MOTE VIETED BY CLINICAL 1362 00:50:32,918 --> 00:50:34,953 OBSERVATIONS THAT SHOWED THAT 1363 00:50:34,953 --> 00:50:37,423 PATIENTS HAVE DEFECTS IN 1364 00:50:37,423 --> 00:50:38,357 METABOLISM ARE HYPOMETABOLIC 1365 00:50:38,357 --> 00:50:39,191 SPECIFICALLY AND HONESTLY I 1366 00:50:39,191 --> 00:50:40,993 DIDN'T REALLY UNDERSTAND WHAT 1367 00:50:40,993 --> 00:50:42,094 THAT MEANT BECAUSE THEY WERE 1368 00:50:42,094 --> 00:50:45,764 REFERRING TO THIS AS THE 1369 00:50:45,764 --> 00:50:47,533 ORGANISMAL LIMP BUT NOBODY COULD 1370 00:50:47,533 --> 00:50:48,934 TELL ME WHAT HAPPENED AT THE 1371 00:50:48,934 --> 00:50:49,134 LEVEL. 1372 00:50:49,134 --> 00:50:50,235 SO THE FLY GAVE US AN 1373 00:50:50,235 --> 00:50:52,438 OPPORTUNITY TO LOOK AT METABOLIC 1374 00:50:52,438 --> 00:50:53,939 CHANGES AT THE CELLULAR LEVEL. 1375 00:50:53,939 --> 00:50:55,607 SO A FORMER VERY TALENTED 1376 00:50:55,607 --> 00:51:00,379 STUDENT IN MY LAB, PURSUED THIS 1377 00:51:00,379 --> 00:51:01,180 METABOLOMIC APPROACH AND I 1378 00:51:01,180 --> 00:51:03,148 SHOWED YOU THIS GRAPH THAT ARE 1379 00:51:03,148 --> 00:51:06,018 CHANGES IN THE GLYCOLYSIS, AND 1380 00:51:06,018 --> 00:51:07,186 IN PYRUVATE SPECIFICALLY 1381 00:51:07,186 --> 00:51:09,655 SUGGESTING THIS THE HIGHER GLIEK 1382 00:51:09,655 --> 00:51:11,557 LYTIC OUTPUT AND THIS WAS ALSO 1383 00:51:11,557 --> 00:51:12,725 IDENTIFIED IN TISSUE CULTURE 1384 00:51:12,725 --> 00:51:17,363 MODELS AS WELL AS IN SOME MOUSE 1385 00:51:17,363 --> 00:51:17,596 MODELS. 1386 00:51:17,596 --> 00:51:20,799 I SHOWED YOU THIS FIGURE BEFORE, 1387 00:51:20,799 --> 00:51:22,568 WE VALIDATED USING QPC R, RANGES 1388 00:51:22,568 --> 00:51:24,970 IN THE LEVELS OR THE EXPRESSION 1389 00:51:24,970 --> 00:51:27,840 OF RATE LIMITING ENZYMES BOTH IN 1390 00:51:27,840 --> 00:51:29,341 GLYCOLYSIS, AS IN FLIES AS WELL 1391 00:51:29,341 --> 00:51:33,379 AS IN PATIENT TISSUES AND IMN, 1392 00:51:33,379 --> 00:51:36,315 SO CONSISTENTLY ACROSS MODELS I 1393 00:51:36,315 --> 00:51:39,618 FOUND AN UPREGULATION OF PFK 1394 00:51:39,618 --> 00:51:40,786 TRANSPORRATE APPROXIMATE 1395 00:51:40,786 --> 00:51:43,856 TENTIALLY WITH ENHANCED 1396 00:51:43,856 --> 00:51:44,156 GLYCOLYSIS. 1397 00:51:44,156 --> 00:51:45,724 HOWEVER, TO OUR SURPRISE WHEN WE 1398 00:51:45,724 --> 00:51:52,731 USE THE LARVA RETURNING ASSAY TO 1399 00:51:52,731 --> 00:51:54,700 MODULATE GLYCOL SIT EAR 1400 00:51:54,700 --> 00:51:55,534 EXPRESSING OR KNOCKING IT DOWN 1401 00:51:55,534 --> 00:51:58,404 WE FOUND THAT IN THE CONTEXT OF 1402 00:51:58,404 --> 00:52:01,707 PDFTREY APPROXIMATE, WE PFK 1403 00:52:01,707 --> 00:52:05,177 RESCUED THE LOCOMOTOR DEFECT, 1404 00:52:05,177 --> 00:52:07,379 EVEN IF THE TRANSCRIPT IS 1405 00:52:07,379 --> 00:52:09,214 HIGHER, IT OVEREXPRESSING PFK 1406 00:52:09,214 --> 00:52:12,084 CAN RESCUE, SUGGESTING TO US 1407 00:52:12,084 --> 00:52:13,886 THAT PFK UPREGULATION AT THE 1408 00:52:13,886 --> 00:52:16,588 TRANSCRIPT LEVEL MUST BE A 1409 00:52:16,588 --> 00:52:17,122 COMPENSATORY MECHANISM, 1410 00:52:17,122 --> 00:52:19,391 POTENTIALLY TO MAKE UP FOR ATP 1411 00:52:19,391 --> 00:52:22,227 PRODUCTION DEFICITS CAUSED BY 1412 00:52:22,227 --> 00:52:22,694 MITOCHONDRIA DEFECTS. 1413 00:52:22,694 --> 00:52:25,831 SO THEN WE WANTED TO KNOW -- OH 1414 00:52:25,831 --> 00:52:29,101 I FORGOT TO MENTION THAT WE, 1415 00:52:29,101 --> 00:52:30,869 PROACH THIS PROBLEM, ALSO WITH I 1416 00:52:30,869 --> 00:52:32,371 DIETARY APPROACH SO IF YOU FEED 1417 00:52:32,371 --> 00:52:34,473 THE HIGH SUGAR DIET TO A NORMAL 1418 00:52:34,473 --> 00:52:36,074 FLY, THAT'S NOT VERY GOOD. 1419 00:52:36,074 --> 00:52:38,811 THAT FLY WILL HAVE AN EFFECT ON 1420 00:52:38,811 --> 00:52:39,845 LOCOMOTOR FUNCTION AS WELL AS 1421 00:52:39,845 --> 00:52:40,679 LIFE SPAN. 1422 00:52:40,679 --> 00:52:42,881 NOT IN A GOOD WAY, BUT IF YOU 1423 00:52:42,881 --> 00:52:46,318 FEED A HIGH SUGAR DIET TO AN ALS 1424 00:52:46,318 --> 00:52:48,654 FLY, THERE WILL BE AN 1425 00:52:48,654 --> 00:52:49,455 IMPROVEMENT IN LOCOMOTOR 1426 00:52:49,455 --> 00:52:50,322 FUNCTION AND INCREASING LIFE 1427 00:52:50,322 --> 00:52:54,526 SPAN AND ACHIEVE THAT AS WELL AS 1428 00:52:54,526 --> 00:52:55,561 GENETICALLY, BY OVEREXPRESSING 1429 00:52:55,561 --> 00:52:56,995 HUMAN GLUCOSE TRANSPORTER JUST 1430 00:52:56,995 --> 00:52:59,665 IN METEOR NEURONS, SO OVERALL IT 1431 00:52:59,665 --> 00:53:01,867 LOOKS LIKE, ENHANCING GLYCOLYSIS 1432 00:53:01,867 --> 00:53:03,035 IN MOTOR NEURON, ALTHOUGH IT MAY 1433 00:53:03,035 --> 00:53:07,940 BE A DESPERATE MEASURE, AND MAY 1434 00:53:07,940 --> 00:53:10,809 NOT BE, PER SE, IT IS HELPING 1435 00:53:10,809 --> 00:53:12,110 SURVIVE AND FUNCTION BETTER. 1436 00:53:12,110 --> 00:53:14,513 SO THEN WE WANTED TO KNOW HOW 1437 00:53:14,513 --> 00:53:16,415 DOES THIS HAPPEN? 1438 00:53:16,415 --> 00:53:20,419 PFK IS AN ENZYME THAT HAS AN ATP 1439 00:53:20,419 --> 00:53:21,954 BINDING DOMAIN THAT CAN BE 1440 00:53:21,954 --> 00:53:23,522 REGULATORY AND ANOTHER 1 THAT'S 1441 00:53:23,522 --> 00:53:23,789 CATALYTIC. 1442 00:53:23,789 --> 00:53:25,257 SO THE FIRST THING WE DID IS WE 1443 00:53:25,257 --> 00:53:27,659 MADE NEW LINES AND WE 1444 00:53:27,659 --> 00:53:30,496 OVEREXPRESSED PFK AND WE FOUND 1445 00:53:30,496 --> 00:53:32,764 THAT THAT CAUSED AN IMPROVEMENT 1446 00:53:32,764 --> 00:53:34,766 IN LOCOMOTOR FUNCTION USING THE 1447 00:53:34,766 --> 00:53:36,268 LARVA TURNING ASSAY BOTH FOR 1448 00:53:36,268 --> 00:53:38,670 WILD-TYPE AS WELL AS MUTANT 1449 00:53:38,670 --> 00:53:38,871 TDP43. 1450 00:53:38,871 --> 00:53:40,772 WE ALSO FOWBD THE SURVIVAL OF 1451 00:53:40,772 --> 00:53:45,511 THE FLY WAS INCREASED BY 1452 00:53:45,511 --> 00:53:46,311 WILD-TYPE PFK OVEREXPRESSION 1453 00:53:46,311 --> 00:53:49,314 BOTH FOR WILD-TYPE AS WELL AWE 1454 00:53:49,314 --> 00:53:49,948 MUSEUM TABT TDP43. 1455 00:53:49,948 --> 00:53:53,952 AND THEN WE WANTED TO MAKE A 1456 00:53:53,952 --> 00:53:56,255 TRANSGENE WHERE THE CATALYTIC 1457 00:53:56,255 --> 00:53:58,690 ATP BINDING SITE IS ALTERED, SO 1458 00:53:58,690 --> 00:54:02,294 THIS FORMER STUDENT IN MY LAB, 1459 00:54:02,294 --> 00:54:05,163 DID SOME MODELING AND WE 1460 00:54:05,163 --> 00:54:07,232 GENERATED TRANSGENIC FLIES THAT 1461 00:54:07,232 --> 00:54:08,600 OVEREXPRESSED THIS MUTANT FORM 1462 00:54:08,600 --> 00:54:11,303 OF PFK THAT IS CATALYTICALLY 1463 00:54:11,303 --> 00:54:13,038 INACTIVE, SO YOU CAN SEE THAT 1464 00:54:13,038 --> 00:54:14,873 WHEN WE OVEREXPRESS THIS FORM OF 1465 00:54:14,873 --> 00:54:18,377 PFK WE DO NOT RESCUE THE 1466 00:54:18,377 --> 00:54:20,312 LOCOMOTOR PHENOTYPE AND WE DO 1467 00:54:20,312 --> 00:54:23,815 NOT RESCUE THE SURVIVAL 1468 00:54:23,815 --> 00:54:24,082 PHENOTYPE. 1469 00:54:24,082 --> 00:54:25,350 CONSISTENT WITH THE IDEA THAT 1470 00:54:25,350 --> 00:54:28,320 THE ATP BINDING SITE OR THE 1471 00:54:28,320 --> 00:54:31,957 CATALYTIC ACTIVITY OF PFK IS 1472 00:54:31,957 --> 00:54:35,060 REQUIRED FOR IMPROVING TDP43 1473 00:54:35,060 --> 00:54:35,527 ASSOCIATED PHENOTYPES. 1474 00:54:35,527 --> 00:54:36,695 AND THEN WE WANTED TO KNOW A 1475 00:54:36,695 --> 00:54:39,331 LITTLE BIT MORE ABOUT HOW MIGHT 1476 00:54:39,331 --> 00:54:43,368 THIS HAPPEN AND WORK IN DANIEL 1477 00:54:43,368 --> 00:54:46,138 RAMS ON LAB AT YALE HAS SHOWN 1478 00:54:46,138 --> 00:54:47,739 MANYIERS AGO THROUGH AN 1479 00:54:47,739 --> 00:54:52,377 INTERESTING ACCIDENT THAT PFK 1480 00:54:52,377 --> 00:54:54,913 FORMS PUNTA UNDER THEOXIDATIVE 1481 00:54:54,913 --> 00:54:55,147 STRESS. 1482 00:54:55,147 --> 00:54:57,149 IT TELLS A NICE STORY ABOUT A 1483 00:54:57,149 --> 00:54:59,251 STUDENT WHO S&P DOING A SCREEN 1484 00:54:59,251 --> 00:55:01,587 AND THE STUDENT LEFT THE COVER 1485 00:55:01,587 --> 00:55:03,155 SLIP ON THE WORM FOR A LITTLE 1486 00:55:03,155 --> 00:55:06,024 LONGER AND THE WORM WAS 1487 00:55:06,024 --> 00:55:07,626 SUGGESTED TOOXIDATIVE STRESS AND 1488 00:55:07,626 --> 00:55:09,461 THAT CAUSED PFK INTO THE 1489 00:55:09,461 --> 00:55:11,096 SYNAPTIC TERMINAL AND IT TURNS 1490 00:55:11,096 --> 00:55:15,133 OUT THIS CAN BE SMOOTH, IN YEAST 1491 00:55:15,133 --> 00:55:17,235 AS WELL, WHERE ANY 1492 00:55:17,235 --> 00:55:17,869 IMMUNOPRECIPITATION HAVE SHOWN 1493 00:55:17,869 --> 00:55:20,305 THAT A NUMBER OF OTHER DPLIEK CO 1494 00:55:20,305 --> 00:55:22,507 LYTIC ENZYMES AND CHAPERONS AND 1495 00:55:22,507 --> 00:55:24,810 EVEN TRANSLATION FACTORS CAN 1496 00:55:24,810 --> 00:55:28,080 ASSOCIATE WITH PFK PUNTA INIEST 1497 00:55:28,080 --> 00:55:29,548 UNDER STRESS SO THEN WE WANTED 1498 00:55:29,548 --> 00:55:30,983 TO KNOW WHETHER THIS MIGHT BE 1499 00:55:30,983 --> 00:55:33,485 HAPPENING IN OUR MODEL, SO THE 1500 00:55:33,485 --> 00:55:36,655 FIRST QUESTION WAS, DOES PFK 1501 00:55:36,655 --> 00:55:39,658 FORM PUNTA, AT THE SYNAPTIC 1502 00:55:39,658 --> 00:55:40,659 TERMINAL UNDER NORMAL CONDITION 1503 00:55:40,659 --> 00:55:42,527 AND THE NEXT QUESTION IS DO 1504 00:55:42,527 --> 00:55:44,262 THOSE PUNTA, ARE THOSE PUNTA 1505 00:55:44,262 --> 00:55:47,733 DIFFERENT IN IS THE ASSUM 1506 00:55:47,733 --> 00:55:49,234 BLEOMYCIN OF PUNTA DIFFERENT IN 1507 00:55:49,234 --> 00:55:52,204 THE CONTEXT OF TDP43 PERHAPS TO 1508 00:55:52,204 --> 00:55:57,342 COMPENSATE FOR ENERGY ENERGY D. 1509 00:55:57,342 --> 00:55:59,678 SO A NEW STUDENT IN MY LAB WITH 1510 00:55:59,678 --> 00:56:01,113 THE NEUROSCIENCE PROGRAM WHO'S 1511 00:56:01,113 --> 00:56:03,849 VERY TALENTED USED A STED HIGH 1512 00:56:03,849 --> 00:56:07,653 RESOLUTION MICROSCOPE TO LOOK AT 1513 00:56:07,653 --> 00:56:10,422 PFK, THIS IS CRISPR KNOCK IN 1514 00:56:10,422 --> 00:56:12,658 WHERE WE GIVE THE PFK IN THE 1515 00:56:12,658 --> 00:56:14,493 ENDOGENOUS CONTEXT SO WE CAN SEE 1516 00:56:14,493 --> 00:56:17,896 PFK IN THE MUSCLE, SO YOU SEE 1517 00:56:17,896 --> 00:56:20,732 THAT GREEN PUNTA THAT'S THE 1518 00:56:20,732 --> 00:56:22,367 MUSCLE, AND THEN THE RED 1519 00:56:22,367 --> 00:56:23,769 STAINING HIGHLIGHTS THE SYNAPTIC 1520 00:56:23,769 --> 00:56:25,404 TERMINAL AND YOU CAN SEE THAT 1521 00:56:25,404 --> 00:56:27,506 WITHIN THE SYNAPTIC TERMINAL, 1522 00:56:27,506 --> 00:56:32,611 THERE ARE PFK PUNTA AND THERE 1523 00:56:32,611 --> 00:56:33,812 UNDER NORMAL CONDITIONS AND WE 1524 00:56:33,812 --> 00:56:35,414 WERE EXCITED ABOUT THAT, AND 1525 00:56:35,414 --> 00:56:39,718 THEN WE STARTED LOOKING AT 1526 00:56:39,718 --> 00:56:42,721 TDP43, EXPRESSING FLIES AND WE 1527 00:56:42,721 --> 00:56:48,326 FIND THAT THERE ARE PUNCTA, SO I 1528 00:56:48,326 --> 00:56:49,828 DON'T HAVE AN ANSWER YOU EXCEPT 1529 00:56:49,828 --> 00:56:50,929 TO SAY THEY LOOK A LITTLE 1530 00:56:50,929 --> 00:56:51,863 DEFINITE AND WE'RE TRYING TO 1531 00:56:51,863 --> 00:56:55,067 FIGURE OUT WHAT MEANS. 1532 00:56:55,067 --> 00:57:01,740 AND I WOULD SAY, IT LOOKS LIKE 1533 00:57:01,740 --> 00:57:05,077 TDP43 ALTERS THE PFK PUNCTA, AND 1534 00:57:05,077 --> 00:57:07,846 THEN WE WENT TO USE AMERIS TO 1535 00:57:07,846 --> 00:57:10,282 LOOK AT THIS AND BEGIN TO 1536 00:57:10,282 --> 00:57:10,982 QUANTIFY THEM. 1537 00:57:10,982 --> 00:57:14,319 SO HERE IS PFK IN A NORMAL 1538 00:57:14,319 --> 00:57:15,921 SYNAPTIC TERMINAL SO ACTUALLY 1539 00:57:15,921 --> 00:57:17,222 THIS IS A QUESTION I HAD FOR 1540 00:57:17,222 --> 00:57:18,690 YOU, IF YOU SEE A DIFFERENCE 1541 00:57:18,690 --> 00:57:19,691 BECAUSE I SEE A DIFFERENCE BUT I 1542 00:57:19,691 --> 00:57:22,327 WOULD LIKE TO KNOW WHAT YOU SEE. 1543 00:57:22,327 --> 00:57:25,464 SO HERE'S A CONTROL, AND HERE'S 1544 00:57:25,464 --> 00:57:35,841 A TDP43 FREE RADICALS, 1545 00:57:36,141 --> 00:57:36,775 EXPRESSING ANIMAL. 1546 00:57:36,775 --> 00:57:37,642 QUANTIFICATION IS IN PROGRESS 1547 00:57:37,642 --> 00:57:39,144 AND IT'S SO NEW THAT I'M NOT 1548 00:57:39,144 --> 00:57:45,617 QUITE SURE WE'RE READY BUT 1549 00:57:45,617 --> 00:57:47,452 VISUALLY, I'LL BE CURIOUS IF YOU 1550 00:57:47,452 --> 00:57:48,854 ALL SEE SOMETHING SO WE CAN TALK 1551 00:57:48,854 --> 00:57:55,260 ABOUT IT BECAUSE I DO SEE 1552 00:57:55,260 --> 00:57:55,660 SOMETHING. 1553 00:57:55,660 --> 00:57:57,395 AND THEN THE LAST SLIDE I HAVE 1554 00:57:57,395 --> 00:57:59,197 IS HOW MIGHT THIS WORK? 1555 00:57:59,197 --> 00:58:02,200 SO THE WHEEL IDEA THAT PFK IS IN 1556 00:58:02,200 --> 00:58:07,973 THE TERMINAL IT MAY BE FORMING 1557 00:58:07,973 --> 00:58:09,107 GLYCOLYTIC PERIODS IN RESPONSE 1558 00:58:09,107 --> 00:58:09,608 TO STRESS. 1559 00:58:09,608 --> 00:58:12,744 WE TESTED THAT IDEA BY USING THE 1560 00:58:12,744 --> 00:58:13,578 FM143 DISCIPLINARY OPTIC ASSAY 1561 00:58:13,578 --> 00:58:15,313 WHICH IS A MEASURE OF SYNAPTIC 1562 00:58:15,313 --> 00:58:16,314 ASSAY IN THE SIGNIFYITOSEIS AND 1563 00:58:16,314 --> 00:58:19,951 YOU CAN SEE IN THE CONTEXT OF 1564 00:58:19,951 --> 00:58:21,186 TDP43, THE SIGNAL IS REDUCED 1565 00:58:21,186 --> 00:58:22,788 BECAUSE THERE ARE REDUCES IN THE 1566 00:58:22,788 --> 00:58:27,659 VESICLE IN THE SIGNIFYITOSEIS. 1567 00:58:27,659 --> 00:58:30,061 OVEREXPRESSION OF PFK RESTORES 1568 00:58:30,061 --> 00:58:34,232 THE SIGNAL IN THOSE MUTANTS 1569 00:58:34,232 --> 00:58:36,201 SUGGESTING THAT PFK 1570 00:58:36,201 --> 00:58:37,169 OVEREXPRESSION RESTORES THE 1571 00:58:37,169 --> 00:58:39,437 ENDOCYTOSIS, AND THIS IS MY LAST 1572 00:58:39,437 --> 00:58:39,671 SLIDE. 1573 00:58:39,671 --> 00:58:41,673 JUST TO SORT OF PUT IT 1574 00:58:41,673 --> 00:58:45,577 ALTOGETHER, WE ARE FOCUSING ON 1575 00:58:45,577 --> 00:58:48,046 THE -- THESE GLYCOLYTIC BODIES. 1576 00:58:48,046 --> 00:58:50,515 WE ARE ISOLATING SYNAPTICSOMES, 1577 00:58:50,515 --> 00:58:52,050 WE ARE DOING PROTEOMICS ON THESE 1578 00:58:52,050 --> 00:58:54,953 TO FIGURE OUT WHAT ARE THE PFK 1579 00:58:54,953 --> 00:58:56,121 ASSEMBLIES, HOW MIGHT THEY BE 1580 00:58:56,121 --> 00:58:57,489 DIFFERENT AND COLLABORATING WITH 1581 00:58:57,489 --> 00:58:59,724 THE COLLEAGUE WHO DOES RYE O EM 1582 00:58:59,724 --> 00:59:01,493 TO LOOK AT CRYO STRUCTURES IN 1583 00:59:01,493 --> 00:59:02,894 THE CHANGES OF THESE MOLECULES 1584 00:59:02,894 --> 00:59:05,664 AT THE SYNAPSE BUT WE DO -- WE 1585 00:59:05,664 --> 00:59:08,466 HAVE OBSERVED AN UPREGULATION 1586 00:59:08,466 --> 00:59:11,970 THAT'S COMPENSATORY. 1587 00:59:11,970 --> 00:59:15,273 THAT IS PROTECTIVE AND THE 1588 00:59:15,273 --> 00:59:17,042 CATALUTTIC ACTIVITY OF PFK IS 1589 00:59:17,042 --> 00:59:17,909 REQUIRED FOR THIS 1590 00:59:17,909 --> 00:59:19,044 NEUROPROTECTIVE ACTIVITY AND IT 1591 00:59:19,044 --> 00:59:22,347 DOES SO BY MITIGATING SYNAPTIC 1592 00:59:22,347 --> 00:59:23,014 VESICLE DEFICITS. 1593 00:59:23,014 --> 00:59:24,449 AGAIN, THE QUESTION STILL 1594 00:59:24,449 --> 00:59:27,352 REMAINS, WHAT DO THESE PFK ASSEM 1595 00:59:27,352 --> 00:59:28,420 MYELINS DO, WHAT ARE THEY, HOW 1596 00:59:28,420 --> 00:59:31,022 DO THEY CHANGE IN THE CONTEXT OF 1597 00:59:31,022 --> 00:59:32,858 TDP 43 AND THIS IS WORK THAT 1598 00:59:32,858 --> 00:59:35,427 WE'RE DURANNENTLY DOING AND HOPE 1599 00:59:35,427 --> 00:59:36,962 TO APPROACH THIS WITH BIOCHEMIST 1600 00:59:36,962 --> 00:59:39,865 RADIOY AS WELL AS CRYOEM 1601 00:59:39,865 --> 00:59:40,565 APPROACHES AND WITH THAT I WANT 1602 00:59:40,565 --> 00:59:42,100 TO THANK YOU FOR YOUR ATTENTION 1603 00:59:42,100 --> 00:59:44,436 AND APOLOGIES I WENT TO THE FULL 1604 00:59:44,436 --> 00:59:44,636 HOUR. 1605 00:59:44,636 --> 00:59:47,606 BUT I DO WANT TO SAY THANKS TO 1606 00:59:47,606 --> 00:59:50,809 THE NEW PEOPLE THAT I RECRUITED 1607 00:59:50,809 --> 00:59:53,445 IN MY LAB AT PENN STATE, VERY 1608 00:59:53,445 --> 00:59:54,446 WONDERFUL COLLABORATORS ACROSS 1609 00:59:54,446 --> 00:59:55,947 THE COUNTRY, ALL THE WAY FROM 1610 00:59:55,947 --> 00:59:58,617 ARIZONA TO PENN STATE AND 1611 00:59:58,617 --> 01:00:01,186 BELGIUM, FORMER STUDENTS IN THE 1612 01:00:01,186 --> 01:00:03,088 LAB WHO CONTRIBUTED TO THIS 1613 01:00:03,088 --> 01:00:03,588 WORK. 1614 01:00:03,588 --> 01:00:06,191 AND THE FUNDING AGENCIES AND, 1615 01:00:06,191 --> 01:00:08,760 PATIENTS AND THEIR FAMILIES AND 1616 01:00:08,760 --> 01:00:10,662 SPECIAL THANKS TO THE NIH WHO 1617 01:00:10,662 --> 01:00:12,163 NOT ONLY FUNDED THIS WORK BUT 1618 01:00:12,163 --> 01:00:14,099 ALSO FUNDED THE TRAINING OF THE 1619 01:00:14,099 --> 01:00:16,902 PEOPLE THAT DID THIS WORK. 1620 01:00:16,902 --> 01:00:18,503 SO THEY WILL CARRY IT FORWARD. 1621 01:00:18,503 --> 01:00:25,343 SO THANK YOU. 1622 01:00:25,343 --> 01:00:26,211 [ APPLAUSE ] 1623 01:00:26,211 --> 01:00:28,013 >> --I DON'T WANT TO CURTAIL ANY 1624 01:00:28,013 --> 01:00:30,682 TALKS WITH THESE BUT QUESTIONS? 1625 01:00:30,682 --> 01:00:34,219 >> SO WONDERFUL TALK, ROSS 1626 01:00:34,219 --> 01:00:35,520 OROXIDATIVE STRESS HAS BEEN 1627 01:00:35,520 --> 01:00:40,358 IMPLICATED IN ALS QUITE A BIT. 1628 01:00:40,358 --> 01:00:44,663 IF YOU TREAT ANTIOXIDANT LIKE 1629 01:00:44,663 --> 01:00:48,466 NAC, HOW MUCH OF THE PATHOLOGY 1630 01:00:48,466 --> 01:00:49,567 CAN YOU REVERSE? 1631 01:00:49,567 --> 01:00:51,403 >> WE TRY TO FEED AD4 WHICH I 1632 01:00:51,403 --> 01:00:54,139 GUESS IS THE SAME THING, THERE 1633 01:00:54,139 --> 01:00:56,708 ARE -- THERE IS A PHENOTYPIC 1634 01:00:56,708 --> 01:01:00,645 RESCUE, HOWEVER, WE WERE NOT 1635 01:01:00,645 --> 01:01:04,482 ABLE TO IDENTIFY ROSS, IN A 1636 01:01:04,482 --> 01:01:04,883 CONSISTENT WAY. 1637 01:01:04,883 --> 01:01:08,320 SO, EVERY TIME WE USED VARIOUS 1638 01:01:08,320 --> 01:01:09,888 APPROACHES TO MEASURE FREE 1639 01:01:09,888 --> 01:01:11,856 RADICALS IN OUR PREPARATIONS, 1640 01:01:11,856 --> 01:01:13,925 THE RESULTS WERE TOO VARIABLE TO 1641 01:01:13,925 --> 01:01:16,194 TELL, AND IN SPEAKS TO EXPERTS, 1642 01:01:16,194 --> 01:01:18,196 IT SEEMS LIKE IT MAY BE 1643 01:01:18,196 --> 01:01:20,265 TRANSIENT AND VERY DIFFICULT, 1644 01:01:20,265 --> 01:01:21,566 TOO, SO I THINK IT'S HAPPENING 1645 01:01:21,566 --> 01:01:23,301 BUT WE HAVEN'T BEEN QUITE ABLE 1646 01:01:23,301 --> 01:01:26,171 TO CAPTURE IT, AND SIMPLY 1647 01:01:26,171 --> 01:01:28,106 FEEDING AN ANTIOXIDANT IS 1648 01:01:28,106 --> 01:01:30,742 PROTECTIVE, BUT I AM NOT SURE 1649 01:01:30,742 --> 01:01:31,943 HOW THAT HAPPENS. 1650 01:01:31,943 --> 01:01:34,312 NTHANK YOU. 1651 01:01:34,312 --> 01:01:34,612 >> THANKS. 1652 01:01:34,612 --> 01:01:36,247 >> SO SINCE YOU ASKED OUR 1653 01:01:36,247 --> 01:01:39,250 OPINION ON THE LAST EXPERIMENT, 1654 01:01:39,250 --> 01:01:39,684 THE QUESTION -- 1655 01:01:39,684 --> 01:01:41,753 >> DO YOU SEE A DIFFERENCE? 1656 01:01:41,753 --> 01:01:43,488 >> WELL, LET ME ASK YOU A 1657 01:01:43,488 --> 01:01:47,292 QUESTION FIRST BEFORE I GIVE AN 1658 01:01:47,292 --> 01:01:47,559 ANSWER. 1659 01:01:47,559 --> 01:01:49,627 HAVE YOU TRIED DOING THAT LIVE 1660 01:01:49,627 --> 01:01:52,263 -- IS THIS -- I'M ASSUME 1661 01:01:52,263 --> 01:01:53,431 THANKSGIVING IS FIXED TISSUE? 1662 01:01:53,431 --> 01:01:54,065 >> YEAH. 1663 01:01:54,065 --> 01:01:55,333 >> HAVE YOU TRIED DOING IT LIVE 1664 01:01:55,333 --> 01:01:57,902 BECAUSE PARTICULARLY IF YOU 1665 01:01:57,902 --> 01:02:01,873 THINK THAT THESE ARE YOU KNOW 1666 01:02:01,873 --> 01:02:03,375 CONDEN SITES AND SO FORTH YOU 1667 01:02:03,375 --> 01:02:05,276 OFTEN GET LARGER EFFECTS ON THE 1668 01:02:05,276 --> 01:02:07,879 DYNAMICS THAN YOU GET ON STEADY 1669 01:02:07,879 --> 01:02:09,914 STATE SO SOMETHING LIKE EITHER 1670 01:02:09,914 --> 01:02:12,117 DOING IT IN LIVE IMAGING OR 1671 01:02:12,117 --> 01:02:14,219 DOING SOMETHING LIKE FRAPP OR 1672 01:02:14,219 --> 01:02:15,754 PHOTO ACTIVATION MAY GIVE YOU A 1673 01:02:15,754 --> 01:02:17,389 MUCH CLEANER ANSWER, MAYBE THE 1674 01:02:17,389 --> 01:02:19,024 DYNAMICS OF THINGS GOING IN AND 1675 01:02:19,024 --> 01:02:23,495 OUT THAT MATTERS MORE THAN 1676 01:02:23,495 --> 01:02:25,563 WHETHER YOU SEE THE AGGREGATES. 1677 01:02:25,563 --> 01:02:28,099 >> WE DID THINK THIS EXPERIMENT. 1678 01:02:28,099 --> 01:02:29,367 >> IT'S VERY HARD I REALIZE. 1679 01:02:29,367 --> 01:02:32,103 >> IT'S VERY HARD, SO AT PENN 1680 01:02:32,103 --> 01:02:35,206 STATE, AT ARIZONA, I USED TO 1681 01:02:35,206 --> 01:02:38,143 HAVE AN UPRIGHT MICROSCOPE WITH 1682 01:02:38,143 --> 01:02:38,510 IMMERSION LENS. 1683 01:02:38,510 --> 01:02:39,210 PENN STATE DOESN'T HAVE THIS, I 1684 01:02:39,210 --> 01:02:40,645 HAVE TO GO TO ANOTHER CAMPUS BUT 1685 01:02:40,645 --> 01:02:41,813 WE WOULD DO THAT. 1686 01:02:41,813 --> 01:02:42,981 MY STUDENT SARAH AND I TALKED 1687 01:02:42,981 --> 01:02:43,848 ABOUT THIS AND SAID WE WOULD 1688 01:02:43,848 --> 01:02:47,886 LIKE TO DO THAT. 1689 01:02:47,886 --> 01:02:49,721 WE WOULD ALSO LIKE TO DO TISSUE 1690 01:02:49,721 --> 01:02:51,022 AND MACROPHAGESSOGRAPHY AND LOOK 1691 01:02:51,022 --> 01:02:52,390 AT THESE STRUCTURES BUT THE 1692 01:02:52,390 --> 01:02:54,459 THING I WE WEREN'T QUITE SURE 1693 01:02:54,459 --> 01:02:55,627 THAT FPK IS THERE BECAUSE WHEN 1694 01:02:55,627 --> 01:02:58,630 YOU USE THE REGULAR CONFOCAL, 1695 01:02:58,630 --> 01:03:00,865 IT'S THERE, BUT WE WANTED A 1696 01:03:00,865 --> 01:03:02,901 HIGHER, WE WANTED TO KNOW USING 1697 01:03:02,901 --> 01:03:05,070 THE HIGH RESOLUTION WHETHER WE 1698 01:03:05,070 --> 01:03:09,808 REALLY DO SEE TRUE FFK PUNCTA, 1699 01:03:09,808 --> 01:03:11,109 THAT'S WHY WE WENT THERE FIRST 1700 01:03:11,109 --> 01:03:13,011 BUT I AGREE WITH YOU THE LIVE 1701 01:03:13,011 --> 01:03:14,479 DYNAMICS OF THIS IS REALLY WHAT 1702 01:03:14,479 --> 01:03:16,781 IT'S GOING TO BE INTERESTING. 1703 01:03:16,781 --> 01:03:17,215 SO, YES, THANKS. 1704 01:03:17,215 --> 01:03:20,185 BUT DO YOU SEE A DIFFERENCE OR 1705 01:03:20,185 --> 01:03:21,419 NO? 1706 01:03:21,419 --> 01:03:23,054 >> I MEAN, I -- RIGHT. 1707 01:03:23,054 --> 01:03:24,255 IN LOOKING AT 1 PICTURE, RIGHT 1708 01:03:24,255 --> 01:03:26,558 WHAT CAN YOU SAY, BUT IF YOU -- 1709 01:03:26,558 --> 01:03:32,831 YOU COULD IMAGINE LOOKING THE 1710 01:03:32,831 --> 01:03:34,365 AMOUNT OF CLUSTERING AT VARIOUS 1711 01:03:34,365 --> 01:03:35,800 LEVELS, THERE'S WAYS TO QUANTIFY 1712 01:03:35,800 --> 01:03:40,438 THAT THAN JUST LOOKING AT 1713 01:03:40,438 --> 01:03:42,640 PUNCTA, FOR LOOKING FOR IMMUNO 1714 01:03:42,640 --> 01:03:43,808 GENERATED AITY AND THINGS LIKE 1715 01:03:43,808 --> 01:03:44,008 THAT. 1716 01:03:44,008 --> 01:03:45,477 >> SO I WILL GIVE YOU WHAT I 1717 01:03:45,477 --> 01:03:48,046 THINK, THIS AREA LOOKS LIKE IT 1718 01:03:48,046 --> 01:03:48,980 HAS FEWER PUNCTA, THERE ARE 1719 01:03:48,980 --> 01:03:51,416 AREAS IN THE SYNAPTIC TERMINAL 1720 01:03:51,416 --> 01:03:54,052 WHERE PFK'S NOT AS MUCH 1721 01:03:54,052 --> 01:03:55,120 LOCALIZED AT THE MEMBRANE AS IT 1722 01:03:55,120 --> 01:03:57,055 IS IN THE CONTROL, YOU KNOW 1723 01:03:57,055 --> 01:03:58,556 QUANTIFICATION IS IN PROGRESS. 1724 01:03:58,556 --> 01:04:00,358 BUT WHEN I SAW THIS, I WAS SUPER 1725 01:04:00,358 --> 01:04:01,960 EXCITED AND WE DID THE BLIND 1726 01:04:01,960 --> 01:04:03,561 CEST OF EVERYBODY IN MY LAB, AND 1727 01:04:03,561 --> 01:04:05,096 SAID DO YOU SEE A DIFFERENCE AND 1728 01:04:05,096 --> 01:04:05,964 EVERYBODY DID WITHOUT TELLING 1729 01:04:05,964 --> 01:04:09,734 THEM WHAT IT WAS, SO, I'M 1730 01:04:09,734 --> 01:04:10,101 HOPEFUL. 1731 01:04:10,101 --> 01:04:11,536 THERE WILL BE A DIFFERENCE WHEN 1732 01:04:11,536 --> 01:04:14,839 WE DO QUANTIFICATION. 1733 01:04:14,839 --> 01:04:15,206 >> THANK YOU. 1734 01:04:15,206 --> 01:04:16,574 FOR THE TALK. 1735 01:04:16,574 --> 01:04:18,276 I WAS WONDERING ABOUT THE 1736 01:04:18,276 --> 01:04:22,480 MECHANISM OF THIS AGGREGATE LIKE 1737 01:04:22,480 --> 01:04:23,982 FORMATION STHERE ANY POST 1738 01:04:23,982 --> 01:04:25,383 TRANSLATIONAL MODICTION IN THE 1739 01:04:25,383 --> 01:04:26,618 PROTEIN THAT IS LIKE TRIGGERED 1740 01:04:26,618 --> 01:04:30,455 BY THE OTHER AGING OR BY STRESS 1741 01:04:30,455 --> 01:04:31,256 OR SOMETHING ELSE? 1742 01:04:31,256 --> 01:04:33,258 WHICH KIND OF SWITCHES THIS FROM 1743 01:04:33,258 --> 01:04:34,359 NORMAL -- 1744 01:04:34,359 --> 01:04:35,960 >> THERE ARE SO POST 1745 01:04:35,960 --> 01:04:37,162 TRANSLATIONAL MODIIVESES AND NEW 1746 01:04:37,162 --> 01:04:38,696 1S ARE COMING ONLINE EVERY DAY 1747 01:04:38,696 --> 01:04:39,664 SO PHOSPHORYLATION WAS THE FIRST 1748 01:04:39,664 --> 01:04:41,900 1 THAT WAS DETECTED AND THEN 1749 01:04:41,900 --> 01:04:43,935 THERE IS ACETYLATION AND THEN 1750 01:04:43,935 --> 01:04:45,170 THERE IS SIMULATION AND THERE IS 1751 01:04:45,170 --> 01:04:50,942 ANOTHER 1 THAT I JUST LEARNED 1752 01:04:50,942 --> 01:04:51,776 ABOUT, CITRONNULATION OR 1753 01:04:51,776 --> 01:04:54,212 SOMETHING LIKE THAT, THERE IS 1754 01:04:54,212 --> 01:04:55,914 MULTIPLE POST TRANSLATIONAL 1755 01:04:55,914 --> 01:04:58,016 MODIFICATION 1756 01:04:58,016 --> 01:04:59,117 MODIFICATIONS THAT PLAY A ROLE 1757 01:04:59,117 --> 01:05:01,186 AND I WILL LET OTHER PEOPLE WHO 1758 01:05:01,186 --> 01:05:02,353 WORK ON STRUCTURES KNOW MORE 1759 01:05:02,353 --> 01:05:02,921 ABOUT THAT. 1760 01:05:02,921 --> 01:05:04,622 >> THANK YOU. 1761 01:05:04,622 --> 01:05:05,190 >> THANKS. 1762 01:05:05,190 --> 01:05:06,591 >> HI, SO VERY INTERESTING TALK, 1763 01:05:06,591 --> 01:05:09,027 I HAVE LIKE COUPLE OF QUESTIONS. 1764 01:05:09,027 --> 01:05:12,463 SO, THE FIRST QUESTION IS, 1765 01:05:12,463 --> 01:05:13,498 REGARDING YOUR FUNCAT 1766 01:05:13,498 --> 01:05:13,932 EXPERIMENT. 1767 01:05:13,932 --> 01:05:18,536 I WAS JUST WONDERING, HOW WAS 1768 01:05:18,536 --> 01:05:20,805 THE EXPERIMENT DONE, WAS LIKE 1769 01:05:20,805 --> 01:05:22,640 THESE IMMUNO ASSAYS SKIPPED IN 1770 01:05:22,640 --> 01:05:23,942 THE MEDIA FOR A PERIOD OF TIME 1771 01:05:23,942 --> 01:05:27,845 OR WAS IT LIKE A POSITIVE -- 1772 01:05:27,845 --> 01:05:28,913 >> WE TRIED DIFFERENT 1773 01:05:28,913 --> 01:05:29,914 APPROACHES, RIGHT BECAUSE YOU 1774 01:05:29,914 --> 01:05:34,552 CAN DO A TIME COURSE, WE REALIZE 1775 01:05:34,552 --> 01:05:36,387 THAT IN ORDER TO REALLY 1776 01:05:36,387 --> 01:05:37,989 UNDERSTAND WHAT'S GOING ON AND 1777 01:05:37,989 --> 01:05:39,390 OPTIMIZE THIS EXPERIMENT, IT'S 1778 01:05:39,390 --> 01:05:40,925 BETTER TO START WITH A CHRONIC 1779 01:05:40,925 --> 01:05:43,194 TREATMENT, SO THE EXPERIMENTS I 1780 01:05:43,194 --> 01:05:45,196 SHOWED YOU ARE THROUGHOUT LARVA 1781 01:05:45,196 --> 01:05:47,265 TIME, SO ABOUT 3, 3 AND HALF 1782 01:05:47,265 --> 01:05:50,535 DAYS OF FEEDING THE ANL, IF YOU 1783 01:05:50,535 --> 01:05:52,503 FEED IT LESS, THE SIGNAL TO 1784 01:05:52,503 --> 01:05:55,473 NOISE RATIO IS NOT AS GOOD, SO 1785 01:05:55,473 --> 01:05:56,908 WHY WOULD OPTIMIZING THIS 1786 01:05:56,908 --> 01:05:58,810 CHRONIC EXPOSURE, WE HOPE TO GO 1787 01:05:58,810 --> 01:06:02,247 BACK TO THOSE MORE ACUTE OR TIME 1788 01:06:02,247 --> 01:06:03,414 COURSE EXPERIMENTS. 1789 01:06:03,414 --> 01:06:05,250 >> YES, SO FOLLOWING THAT UP, SO 1790 01:06:05,250 --> 01:06:07,318 WHAT YOU ARE SIGNAL IS SHOWING 1791 01:06:07,318 --> 01:06:10,188 THAT IN THESE MUTANT MODELS, YOU 1792 01:06:10,188 --> 01:06:13,758 HAVE LESS OF NEW PROTEINS BEING 1793 01:06:13,758 --> 01:06:13,992 MADE? 1794 01:06:13,992 --> 01:06:14,292 >> YEAH. 1795 01:06:14,292 --> 01:06:16,728 >> AND ALSO YOU ARE SHOWING SOME 1796 01:06:16,728 --> 01:06:19,264 KIND OF ENERGY DEFICIT SO YOU'RE 1797 01:06:19,264 --> 01:06:20,298 BASICALLY SAYING THERE'S A 1798 01:06:20,298 --> 01:06:22,000 MITOCHONDRIA DEFECT BUT HAVE YOU 1799 01:06:22,000 --> 01:06:24,168 LIKE CHECKED IT DIRECTLY? 1800 01:06:24,168 --> 01:06:26,004 >> YOU MEAN LIKE FUNCTIONALLY 1801 01:06:26,004 --> 01:06:27,939 WITH THE MITOCHONDRIA 1802 01:06:27,939 --> 01:06:28,306 FUNCTIONAL? 1803 01:06:28,306 --> 01:06:29,574 >> FOR COMPLEX 1 FUNCTION OR -- 1804 01:06:29,574 --> 01:06:30,608 >> I WE DON'T KNOW HOW TO DO 1805 01:06:30,608 --> 01:06:31,909 THAT, WE WOULD HAVE TO WORK WITH 1806 01:06:31,909 --> 01:06:33,878 -- WE DID THINK ABOUT THAT TO 1807 01:06:33,878 --> 01:06:34,712 LOOK AT COMPLEX 1 FUNCTION AND 1808 01:06:34,712 --> 01:06:36,314 THE EXPECTATION WOULD BE THAT 1809 01:06:36,314 --> 01:06:41,052 IT'S ALTERED JUST BECAUSE ALTER 1810 01:06:41,052 --> 01:06:42,053 INDEED THE NEURODEGENERATIVE 1811 01:06:42,053 --> 01:06:44,188 DISEASES BECAUSE OF THE COMPLEX 1812 01:06:44,188 --> 01:06:45,390 1 COMPONENT. 1813 01:06:45,390 --> 01:06:47,925 WE HAVE DONE TMRE LOOKING AT 1814 01:06:47,925 --> 01:06:49,127 MEMBRANE POTENTIAL OF 1815 01:06:49,127 --> 01:06:49,994 MITOCHONDRIA AND THAT'S ALTERED 1816 01:06:49,994 --> 01:06:51,829 SO WE KNOW THERE'S ALTERATIONS 1817 01:06:51,829 --> 01:06:53,698 ALSO THE STRUCTURE AT DATA, I 1818 01:06:53,698 --> 01:06:56,968 KIND OF BRUSHED OVER THAT BUT 1819 01:06:56,968 --> 01:06:57,902 ULTRA STRUCTURALLY THE 1820 01:06:57,902 --> 01:06:58,836 MITOCHONDRIA ARE REALLY WEIRD, 1821 01:06:58,836 --> 01:07:00,838 LIKE IN THE MUSCLE WE ACTUALLY 1822 01:07:00,838 --> 01:07:03,074 SEE SOME MITOCHONDRIA THAT ARE 1823 01:07:03,074 --> 01:07:05,043 LOOK LIKE THEY'RE ENGULFING 1824 01:07:05,043 --> 01:07:07,712 RIBOSOMES SO THEY'RE JUST VERY 1825 01:07:07,712 --> 01:07:09,781 WEIRD BUT THE EXPECTATION WILL 1826 01:07:09,781 --> 01:07:12,150 BE THAT THE FUNCTIONAL ASSAY 1827 01:07:12,150 --> 01:07:13,785 WOULD DEVELOP A DEFECT. 1828 01:07:13,785 --> 01:07:15,553 >> OKAY, SO SORRY 1 LAST 1829 01:07:15,553 --> 01:07:16,187 QUESTION, -- 1830 01:07:16,187 --> 01:07:16,888 >> SO ACTUALLY IN THE INTEREST 1831 01:07:16,888 --> 01:07:19,390 OF TIME, THERE WILL BE 1832 01:07:19,390 --> 01:07:20,158 OPPORTUNITIES FOR FURTHER 1833 01:07:20,158 --> 01:07:22,894 DISCUSSION OUTSIDE FOR THE 1834 01:07:22,894 --> 01:07:23,161 RECEPTION. 1835 01:07:23,161 --> 01:07:25,096 I WANT JUST TAKE 1 LAST QUESTION 1836 01:07:25,096 --> 01:07:26,264 AND THEN WE RESILIENCE TIRE TO 1837 01:07:26,264 --> 01:07:28,032 THE RECEPTION FOR FURTHER 1838 01:07:28,032 --> 01:07:29,701 QUESTIONS AND DISCUSSION. 1839 01:07:29,701 --> 01:07:30,435 >> OKAY, THANKS. 1840 01:07:30,435 --> 01:07:32,403 >> DON'T LIVE LEAVE, I CAN 1841 01:07:32,403 --> 01:07:33,905 ANSWER YOUR QUESTION LATER. 1842 01:07:33,905 --> 01:07:35,406 >> THIS IS AICH SIMPLE 1 I 1843 01:07:35,406 --> 01:07:36,307 THINK, YOU SHOWED US A NUMBER OF 1844 01:07:36,307 --> 01:07:38,376 CASE WHERE IS YOU HAVE REDUCED 1845 01:07:38,376 --> 01:07:39,344 PROTEIN SYNTHESIS, USING FUN CAT 1846 01:07:39,344 --> 01:07:40,978 SYSTEM OR JUST LOOKINGA THE 1847 01:07:40,978 --> 01:07:43,715 STEADY STATE LEVELS, AT THE 1848 01:07:43,715 --> 01:07:45,149 SYNAPSES, MY QUESTION IS, DO YOU 1849 01:07:45,149 --> 01:07:46,918 REALLY KNOW THAT THAT THIS IS 1850 01:07:46,918 --> 01:07:48,453 REDUCED TRANSLATION, ARE THE 1851 01:07:48,453 --> 01:07:50,054 RNAs REDUCED AT THOSE PLACES? 1852 01:07:50,054 --> 01:07:52,357 COULD IT BE A TRANSPORTER OR 1853 01:07:52,357 --> 01:07:54,092 EVEN A VERY INDIRECT 1854 01:07:54,092 --> 01:07:55,360 TRANSCRIPTION DEFECT RATHER THAN 1855 01:07:55,360 --> 01:07:56,294 A TRANSLATION PROBLEM IN. 1856 01:07:56,294 --> 01:07:57,795 >> WE DON'T THINK IT'S A 1857 01:07:57,795 --> 01:08:00,398 TRANSCRIPTION DEFECT BECAUSE 1858 01:08:00,398 --> 01:08:07,605 WE'VE DONE TRANSCRIPTOMICS IN 1859 01:08:07,605 --> 01:08:08,706 PARALLEL, THERE DEFINITELY A 1860 01:08:08,706 --> 01:08:09,607 TRANSPORTATION DEFICIT. 1861 01:08:09,607 --> 01:08:11,376 THOSE 2 MIGHT BE COUPLED YOU ARE 1862 01:08:11,376 --> 01:08:12,410 ABSOLUTELY RIGHT. 1863 01:08:12,410 --> 01:08:15,113 THERE ARE OTHERS IN THE FIELD 1864 01:08:15,113 --> 01:08:16,948 WHO HAVE USED MICROFLUIDICS AND 1865 01:08:16,948 --> 01:08:19,484 LOOK AT LOCAL TRANSLATION 1866 01:08:19,484 --> 01:08:20,785 DEFECTS CAUSED BY TDP43 AND THEY 1867 01:08:20,785 --> 01:08:22,520 SHOWED THAT IN THAT SYSTEM WHERE 1868 01:08:22,520 --> 01:08:24,655 YOU CAN LOOK LOCALLY, SO IN OUR 1869 01:08:24,655 --> 01:08:26,724 SYSTEM, I CANNOT DISCONNECT THE 1870 01:08:26,724 --> 01:08:28,359 TRANSPORT FROM TRANSLATION BUT I 1871 01:08:28,359 --> 01:08:29,560 DO KNOW THAT TRANSCRIPTION IS 1872 01:08:29,560 --> 01:08:33,331 NOT ALTERED FOR THIS. 1873 01:08:33,331 --> 01:08:33,631 >> THANKS. 1874 01:08:33,631 --> 01:08:34,732 >> SO THANK YOU VERY MUCH AND 1875 01:08:34,732 --> 01:08:37,635 JUST AS A REMINDER, WELL IS A 1876 01:08:37,635 --> 01:08:38,803 RECEPTION OWZ, A SMALL RECEPTION 1877 01:08:38,803 --> 01:08:40,438 SO IF YOU HAVE FURTHER QUESTIONS 1878 01:08:40,438 --> 01:08:41,939 OR WANT TO DISCUSS OTHER THINGS 1879 01:08:41,939 --> 01:08:45,042 PLEASE COME TO THAT AND HELP ME 1880 01:08:45,042 --> 01:08:46,878 THANK DANIELA FOR THE WONDERFUL 1881 01:08:46,878 --> 01:08:47,712 TALK TODAY. 1882 01:08:47,712 --> 01:08:51,416 >> THANK YOU. 1883 01:08:51,416 --> 01:09:01,659 [ APPLAUSE ]