1 00:00:06,055 --> 00:00:07,257 >> GOOD AFTERNOON, EVERYBODY. 2 00:00:07,257 --> 00:00:09,659 THANK YOU SO MUCH FOR COMING. 3 00:00:09,659 --> 00:00:11,961 I CANNOT TELL YOU, EVEN NOW, SO 4 00:00:11,961 --> 00:00:14,063 MANY YEARS OUT, HOW WONDERFUL IT 5 00:00:14,063 --> 00:00:17,233 IS TO SEE REAL HUMAN BEINGS IN A 6 00:00:17,233 --> 00:00:17,967 ROOM. 7 00:00:17,967 --> 00:00:19,068 BUT WELCOME AS WELL TO THOSE OF 8 00:00:19,068 --> 00:00:24,040 YOU WHO HAVE JOINED US ON 9 00:00:24,040 --> 00:00:24,874 VIDEOCAST. 10 00:00:24,874 --> 00:00:28,044 OUR SPEAKER TODAY IS DR. DANIEL 11 00:00:28,044 --> 00:00:28,278 CHERTOW. 12 00:00:28,278 --> 00:00:30,580 DR. CHERTOW RECEIVED HIS 13 00:00:30,580 --> 00:00:32,448 UNDERGRADUATE DEGREE IN 14 00:00:32,448 --> 00:00:34,083 SOCIOLOGY FROM BOSTON COLLEGE 15 00:00:34,083 --> 00:00:36,619 AND HIS MEDICAL DEGREE FROM 16 00:00:36,619 --> 00:00:37,387 NORTHWESTERN UNIVERSITY. 17 00:00:37,387 --> 00:00:39,355 HE THEN WENT ON TO GET A 18 00:00:39,355 --> 00:00:41,658 MASTER'S DEGREE IN PUBLIC HEALTH 19 00:00:41,658 --> 00:00:44,260 FROM THE UNIVERSITY OF 20 00:00:44,260 --> 00:00:47,864 CALIFORNIA AT BERKELEY. 21 00:00:47,864 --> 00:00:51,167 HE PURSUED RESIDENCY TRAINING IN 22 00:00:51,167 --> 00:00:54,470 INTERNAL MEDICINE AT UCSD, AND 23 00:00:54,470 --> 00:00:58,074 THEN HE WENT IN A SOMEWHAT 24 00:00:58,074 --> 00:01:01,978 UNUSUAL MODE TO BECOME AN 25 00:01:01,978 --> 00:01:03,212 EPIDEMIC INTELLIGENCE SERVICE 26 00:01:03,212 --> 00:01:05,281 OFFICER THROUGH THE CDC. 27 00:01:05,281 --> 00:01:08,785 HE THEN WENT ON TO A FELLOWSHIP 28 00:01:08,785 --> 00:01:11,187 IN CRITICAL CARE MEDICINE HERE 29 00:01:11,187 --> 00:01:14,057 AT THE NIH. 30 00:01:14,057 --> 00:01:17,093 AND IN INFECTIOUS DISEASES AT 31 00:01:17,093 --> 00:01:18,728 JOHNS HOPKINS UNIVERSITY. 32 00:01:18,728 --> 00:01:20,496 DR. CHERTOW HAS WON MANY, MANY, 33 00:01:20,496 --> 00:01:21,030 MANY AWARDS. 34 00:01:21,030 --> 00:01:24,100 I'M JUST GOING TO READ OFF A FEW 35 00:01:24,100 --> 00:01:24,867 OF THEM. 36 00:01:24,867 --> 00:01:28,905 THE UNITED STATES PUBLIC HEALTH 37 00:01:28,905 --> 00:01:29,472 SERVICE MERITORIOUS SERVICE 38 00:01:29,472 --> 00:01:32,875 MEDAL, THE UNITED STATES PUBLIC 39 00:01:32,875 --> 00:01:36,045 HEALTH SERVICE PRESIDENTIAL UNIT 40 00:01:36,045 --> 00:01:37,447 CITATION, THE LASKER BLOOMBERG 41 00:01:37,447 --> 00:01:38,648 PUBLIC SERVICE AWARD, THE LIST 42 00:01:38,648 --> 00:01:41,417 REALLY GOES ON AND ON. 43 00:01:41,417 --> 00:01:50,059 AND REALLY IS AN EMBLEM TO HIS 44 00:01:50,059 --> 00:01:53,930 PASSION FOR TAKING PATHOGENESIS 45 00:01:53,930 --> 00:01:55,198 INFORMATION, INFECTIOUS DISEASE 46 00:01:55,198 --> 00:01:56,232 BASIC INFORMATION, AND BRINGING 47 00:01:56,232 --> 00:01:58,935 THEM TO BEAR ON THE HEALTH AND 48 00:01:58,935 --> 00:02:00,803 WELL-BEING OF THE FELLOW PEOPLE 49 00:02:00,803 --> 00:02:03,306 WHO INHABIT THE PLANET WITH HIM. 50 00:02:03,306 --> 00:02:08,878 WHEN I SAY PLANET, I MEAN 51 00:02:08,878 --> 00:02:13,416 GLOBAL, INTERNATIONAL PLANET. 52 00:02:13,416 --> 00:02:14,884 SO, DR. CHERTOW'S TRANSLATIONAL 53 00:02:14,884 --> 00:02:16,419 RESEARCH PROGRAM EMPLOYS 54 00:02:16,419 --> 00:02:18,955 ADVANCED ANIMAL MODELS AND 55 00:02:18,955 --> 00:02:20,490 DETAILED NATURAL HISTORY STUDIES 56 00:02:20,490 --> 00:02:23,659 IN PEOPLE, TO IMPROVE THE 57 00:02:23,659 --> 00:02:25,395 UNDERSTANDING OF THE 58 00:02:25,395 --> 00:02:28,131 PATHOPHYSIOLOGY AND THE 59 00:02:28,131 --> 00:02:29,465 MOLECULAR PATHOGENESIS OF 60 00:02:29,465 --> 00:02:31,968 EMERGING VIRAL INFECTIONS, AND 61 00:02:31,968 --> 00:02:35,004 UP UNTIL RECENTLY THAT INCLUDED 62 00:02:35,004 --> 00:02:40,376 INFLUENZA A, EBOLA, AND ZIKA 63 00:02:40,376 --> 00:02:42,345 VIRUSES BUT AS YOU'LL HEAR WE 64 00:02:42,345 --> 00:02:45,648 CAN ADD SARS-COV-2 TO THAT LIST. 65 00:02:45,648 --> 00:02:49,786 AND THE INTENT IS REALLY TO USE, 66 00:02:49,786 --> 00:02:51,754 AGAIN, THIS VERY BASIC 67 00:02:51,754 --> 00:02:56,692 INFORMATION TO GUIDE CLINICAL 68 00:02:56,692 --> 00:02:57,427 MANAGEMENT, DIAGNOSIS, 69 00:02:57,427 --> 00:02:58,761 UNDERSTANDING OF HUMAN DISEASE. 70 00:02:58,761 --> 00:03:03,599 SO HIS TALK TODAY IS ENTITLED 71 00:03:03,599 --> 00:03:06,769 "REVIVING POSTMORTEM STUDIES TO 72 00:03:06,769 --> 00:03:09,172 CLARIFY INFECTIOUS DISEASES 73 00:03:09,172 --> 00:03:09,505 PATHOGENESIS." 74 00:03:09,505 --> 00:03:11,340 WELCOME, DR. CHERTOW. 75 00:03:11,340 --> 00:03:16,279 [APPLAUSE] 76 00:03:16,279 --> 00:03:18,481 >> THANK YOU, DR. SCHOR, FOR THE 77 00:03:18,481 --> 00:03:19,415 VERY KIND INTRODUCTION. 78 00:03:19,415 --> 00:03:22,084 IT IS MY ABSOLUTE PLEASURE TO BE 79 00:03:22,084 --> 00:03:22,552 HERE. 80 00:03:22,552 --> 00:03:24,053 SO, GOOD AFTERNOON. 81 00:03:24,053 --> 00:03:26,989 AND THANK YOU ALL FOR JOINING IN 82 00:03:26,989 --> 00:03:31,160 PERSON AND ONLINE TODAY. 83 00:03:31,160 --> 00:03:33,095 AS DR. SCHOR MENTION, THE TITLE 84 00:03:33,095 --> 00:03:34,864 IS REVIVING POSTMORTEM STUDIES 85 00:03:34,864 --> 00:03:40,102 TO CLARIFY INFECTIOUS DISEASES 86 00:03:40,102 --> 00:03:40,436 PATHOGENESIS. 87 00:03:40,436 --> 00:03:46,142 I SAY REVIVING BECAUSE AUTOPSIES 88 00:03:46,142 --> 00:03:48,544 HAVE LONG BEEN A TOOL FOR 89 00:03:48,544 --> 00:03:50,980 ESTABLISHING CLINICAL AND 90 00:03:50,980 --> 00:03:53,349 PATHOLOGICAL CORRELATES OF 91 00:03:53,349 --> 00:03:54,116 DISEASE. 92 00:03:54,116 --> 00:03:57,286 PICTURED HER IS SIR WILLIAM 93 00:03:57,286 --> 00:04:00,890 OSLER PERFORMING A BRAIN AUTOPSY 94 00:04:00,890 --> 00:04:02,558 IN THE LATE 1800s AT A 95 00:04:02,558 --> 00:04:03,125 PHILADELPHIA HOSPITAL, ALSO 96 00:04:03,125 --> 00:04:04,660 RECOGNIZED AT THAT TIME 97 00:04:04,660 --> 00:04:07,497 INVESTIGATING CAUTIONS OF DEATH 98 00:04:07,497 --> 00:04:10,666 DIRECTLY INFORMS DISEASE 99 00:04:10,666 --> 00:04:13,202 PREVENTION AND TREATMENT. 100 00:04:13,202 --> 00:04:16,806 IN RECENT DECADES, HOWEVER, THE 101 00:04:16,806 --> 00:04:20,276 FREQUENCY OF AUTOPSIES HAS 102 00:04:20,276 --> 00:04:20,877 SIGNIFICANTLY DECLINED DESPITE 103 00:04:20,877 --> 00:04:22,612 INCREASED AVAILABILITY OF MANY 104 00:04:22,612 --> 00:04:26,082 NO POWERFUL TOOLS TO INVESTIGATE 105 00:04:26,082 --> 00:04:28,651 DISEASE PATHOGENESIS WITHIN 106 00:04:28,651 --> 00:04:29,051 TISSUES. 107 00:04:29,051 --> 00:04:31,287 IN APRIL OF 2020, THROUGH APRIL 108 00:04:31,287 --> 00:04:34,557 OF 2021, DURING THE FIRST YEAR 109 00:04:34,557 --> 00:04:37,393 OF THE COVID PANDEMIC, WE, AND I 110 00:04:37,393 --> 00:04:38,628 MEAN A LARGE CONSORTIUM WHICH 111 00:04:38,628 --> 00:04:41,797 YOU'LL HEAR ABOUT TODAY, 112 00:04:41,797 --> 00:04:45,034 PERFORMED AUTOPSY STUDY OF 44 113 00:04:45,034 --> 00:04:47,370 FATAL COVID-19 CASES RECRUITED 114 00:04:47,370 --> 00:04:48,905 FROM REGIONAL HOSPITALS, AND 115 00:04:48,905 --> 00:04:51,040 THESE AUTOPSIES WERE PERFORMED 116 00:04:51,040 --> 00:04:52,608 AT THE NIH CLINICAL CENTER, FOR 117 00:04:52,608 --> 00:04:55,144 PURPOSE OF LEVERAGING SOME OF 118 00:04:55,144 --> 00:04:58,214 THESE NEW TOOLS TO INFORM 119 00:04:58,214 --> 00:04:59,749 COVID-19 PATHOGENESIS, A NEW 120 00:04:59,749 --> 00:05:01,851 DISEASE THAT HAD SHOWN UP ON THE 121 00:05:01,851 --> 00:05:03,052 SCENE. 122 00:05:03,052 --> 00:05:04,554 THIS WORK WAS ACCOMPLISHED WITH 123 00:05:04,554 --> 00:05:06,656 CLINICAL COLLEAGUES AT THE 124 00:05:06,656 --> 00:05:09,492 UNIVERSITY OF MARYLAND, OUR 125 00:05:09,492 --> 00:05:10,793 CLOSE COLLEAGUES AT THE NATIONAL 126 00:05:10,793 --> 00:05:13,429 CANCER INSTITUTE, ONE OF THOSE 127 00:05:13,429 --> 00:05:16,165 IS HERE TODAY, DR. HEWETT, 128 00:05:16,165 --> 00:05:17,800 CO-COLLABORATOR AND LEAD ON 129 00:05:17,800 --> 00:05:23,606 NUMBER OF THESE STUDIES, MANY 130 00:05:23,606 --> 00:05:27,043 COLLABORATORS I HOPE TO 131 00:05:27,043 --> 00:05:27,343 HIGHLIGHT. 132 00:05:27,343 --> 00:05:31,280 THE OBJECTIVES WERE TO DEFINE 133 00:05:31,280 --> 00:05:32,481 SARS-COV-2 DISTRIBUTION, 134 00:05:32,481 --> 00:05:33,149 CELLULAR TROPISM, PERSISTENCE 135 00:05:33,149 --> 00:05:36,152 AND EVOLUTION IN THE HUMAN BODY, 136 00:05:36,152 --> 00:05:38,821 INCLUDING THE BRAIN, AND TO USE 137 00:05:38,821 --> 00:05:40,957 STANDARD AND EMERGING TOOLS TO 138 00:05:40,957 --> 00:05:43,459 CHARACTERIZE THE CELLULAR AND 139 00:05:43,459 --> 00:05:47,496 MOLECULAR RESPONSES TO INFECTION 140 00:05:47,496 --> 00:05:48,030 WITHIN TISSUES. 141 00:05:48,030 --> 00:05:50,666 TO ACCOMPLISH THESE AIMS, WE 142 00:05:50,666 --> 00:05:54,503 PROCURED TISSUES FROM ALL MAJOR 143 00:05:54,503 --> 00:05:56,372 ORGANS IN 44 FATAL COVID-19 144 00:05:56,372 --> 00:05:57,907 CASES WITH EXTENSIVE BRAIN 145 00:05:57,907 --> 00:06:01,177 SAMPLING IN 11 OF THESE CASES. 146 00:06:01,177 --> 00:06:04,714 WE PRESERVED GREATER THAN 10,000 147 00:06:04,714 --> 00:06:06,248 SPECIMENS ACROSS 85 DISTINCT 148 00:06:06,248 --> 00:06:10,519 ANATOMIC SITES AND BODY FLUIDS. 149 00:06:10,519 --> 00:06:12,154 ADJACENT SPECIMENS WERE 150 00:06:12,154 --> 00:06:15,958 PRESERVED IN RNA LATER, FOR 151 00:06:15,958 --> 00:06:19,128 MOLECULAR STUDIES SUCH AS PCR 152 00:06:19,128 --> 00:06:21,030 AND SEQUENCING, FLASH FROZEN FOR 153 00:06:21,030 --> 00:06:25,635 CELL CULTURE SHOW PRESENCE OR 154 00:06:25,635 --> 00:06:27,503 ABSENCE OF VIABLE VIRUS, AND 155 00:06:27,503 --> 00:06:30,906 FIXED FOR DOWN EXTREME 156 00:06:30,906 --> 00:06:33,075 MICROSCOPY-BASED ANALYSES. 157 00:06:33,075 --> 00:06:40,950 OUR COHORT WAS DIVERSE ACROSS 158 00:06:40,950 --> 00:06:42,151 AGE, SEX, RACE, ETHNICITY. 159 00:06:42,151 --> 00:06:44,253 AT THIS STAGE OF THE PANDEMIC NO 160 00:06:44,253 --> 00:06:46,656 ONE WAS VACCINATED. 161 00:06:46,656 --> 00:06:50,259 THE WA-1, WILD TYPE STRAIN OF 162 00:06:50,259 --> 00:06:51,360 SARS-COV-2 WAS CIRCULATING, KEEP 163 00:06:51,360 --> 00:06:54,030 THOSE THINGS IN MIND US A THINK 164 00:06:54,030 --> 00:06:57,066 ABOUT POTENTIAL GENERALIZABILITY 165 00:06:57,066 --> 00:06:58,367 OF OUR FINDINGS. 166 00:06:58,367 --> 00:07:02,204 77% OF OUR PATIENTS HAD TWO OR 167 00:07:02,204 --> 00:07:05,274 MORE COMORBID CONDITIONS WITH 168 00:07:05,274 --> 00:07:07,810 CARDIOVASCULAR DISEASE, CHRONIC 169 00:07:07,810 --> 00:07:09,445 RESPIRATORY DISEASE, AND 170 00:07:09,445 --> 00:07:10,312 DIABETES, HYPERTENSION, AND 171 00:07:10,312 --> 00:07:14,817 OBESITY BEING THE MOST PREVALENT 172 00:07:14,817 --> 00:07:15,317 CO-MORBIDITIES. 173 00:07:15,317 --> 00:07:18,421 AS SHOWN IN THIS HEAT MAP OF ALL 174 00:07:18,421 --> 00:07:20,890 RNA LEVELS ACROSS TISSUES IN 175 00:07:20,890 --> 00:07:26,295 PATIENTS, WE DETECTED A HIGH 176 00:07:26,295 --> 00:07:29,565 BURDEN OF SARS-COV-2 IN THE 177 00:07:29,565 --> 00:07:33,969 DIGESTIVE TRACT, UPPER AND LOWER 178 00:07:33,969 --> 00:07:35,071 RESPIRATORY TRACT, PREDOMINANTLY 179 00:07:35,071 --> 00:07:38,274 EARLY AFTER ILLNESS, ALSO 180 00:07:38,274 --> 00:07:39,675 IMPORTANTLY OBSERVED PERSISTENCE 181 00:07:39,675 --> 00:07:41,644 OF VIRAL RNA ACROSS THESE 182 00:07:41,644 --> 00:07:46,682 TISSUES FOR MONTHS AFTER ILLNESS 183 00:07:46,682 --> 00:07:46,882 ONSET. 184 00:07:46,882 --> 00:07:49,518 IN WORK LED BY BLAKE WARNER AND 185 00:07:49,518 --> 00:07:53,122 COLLEAGUES IN THE DENTAL 186 00:07:53,122 --> 00:07:54,356 INSTITUTE, THE CELLULAR 187 00:07:54,356 --> 00:08:00,229 DISTRIBUTION OF SARS-COV-2 IN 188 00:08:00,229 --> 00:08:02,098 SALIVARY GLANDS IS SHOWN ON THE 189 00:08:02,098 --> 00:08:04,366 LEFT IMAGE, AND FREQUENT FRIENDS 190 00:08:04,366 --> 00:08:05,835 OF ACTIVATED T- AND B-CELLS 191 00:08:05,835 --> 00:08:07,903 SHOWN IN THE RIGHT IMAGE 192 00:08:07,903 --> 00:08:09,638 PROVIDED INSIGHT INTO THE 193 00:08:09,638 --> 00:08:11,741 MECHANISMS CONTRIBUTING TO THE 194 00:08:11,741 --> 00:08:15,878 LOSS OF TASTE IN COVID-19 195 00:08:15,878 --> 00:08:17,213 PATIENTS. 196 00:08:17,213 --> 00:08:18,180 MOVING DOWN THE RESPIRATORY 197 00:08:18,180 --> 00:08:20,583 TRACT, IN WORK LED BY JENNY 198 00:08:20,583 --> 00:08:23,018 TINGE AND HER COLLEAGUES AT 199 00:08:23,018 --> 00:08:25,087 UNIVERSITY OF NORTH CAROLINA, 200 00:08:25,087 --> 00:08:28,891 THE PRIMARY CELL DEATH EVENT IN 201 00:08:28,891 --> 00:08:31,961 SARS-COV-2 INFECTED AIRWAY 202 00:08:31,961 --> 00:08:35,698 EPITHELIAL WAS WAS DETERMINED TO 203 00:08:35,698 --> 00:08:37,466 BE NECROPTOSIS, A REPRESENTATIVE 204 00:08:37,466 --> 00:08:39,969 IMAGE AMONG EIGHT AUTOPSY CASES 205 00:08:39,969 --> 00:08:44,006 THAT THEY EVALUATED, SHOWING 206 00:08:44,006 --> 00:08:46,742 CO-LOCALIZATION OF SARS-COV-2 207 00:08:46,742 --> 00:08:47,943 NUCLEAR PROTEIN WITH 208 00:08:47,943 --> 00:08:49,945 PHOSPHORYLATED MLK, A LATE 209 00:08:49,945 --> 00:08:54,517 PROTEIN IN THE QUITE 210 00:08:54,517 --> 00:08:55,184 PRO-INFLAMMATORY NECROPTOTIC 211 00:08:55,184 --> 00:08:57,586 CELL-DEATH PATHWAY. 212 00:08:57,586 --> 00:09:03,492 REMAINING IN THE AIRWAYS IN WORK 213 00:09:03,492 --> 00:09:05,427 LED BY RICK BUCHEY AT UNIVERSITY 214 00:09:05,427 --> 00:09:09,498 OF NORTH CAROLINA, MUCUS 215 00:09:09,498 --> 00:09:19,608 PLUGGING DETECTED IN 60% OF 970 216 00:09:19,608 --> 00:09:25,447 SMALL AIRWAYS ACROSS 15 AUTOPSY 217 00:09:25,447 --> 00:09:30,553 CASES. 218 00:09:30,553 --> 00:09:32,755 MUC 5B RATHER THAN 5 AS 219 00:09:32,755 --> 00:09:34,590 PREDOMINATED, SHOWN ON THE RIGHT 220 00:09:34,590 --> 00:09:36,592 IMAGE, REPRESENTATIVE IMAGE ON 221 00:09:36,592 --> 00:09:38,527 THE RIGHT. 222 00:09:38,527 --> 00:09:41,697 PROVIDING NOVEL INSIGHTS INTO 223 00:09:41,697 --> 00:09:42,798 THE CONTRIBUTION OF MUCOUS 224 00:09:42,798 --> 00:09:45,868 ACCUMULATION OF PLUGGING OF 225 00:09:45,868 --> 00:09:49,438 SMALL AIRWAYS TO RESPIRATORY 226 00:09:49,438 --> 00:09:50,506 INSUFFICIENCY AND FAILURE IN 227 00:09:50,506 --> 00:09:51,674 SEVERE COVID-19. 228 00:09:51,674 --> 00:09:54,410 WITHIN THE LUNG PARENCHYMA 229 00:09:54,410 --> 00:09:58,747 ITSELF, WE AND OTHERS HAVE SHOWN 230 00:09:58,747 --> 00:10:01,050 THAT DIFFUSE ALVEOLAR DAMAGE IN 231 00:10:01,050 --> 00:10:02,818 THE EXUDATIVE PHASE IN PANEL A 232 00:10:02,818 --> 00:10:05,354 ON THE LEFT AND PROLIFERATIVE 233 00:10:05,354 --> 00:10:07,423 PHASE, LATER PHASE IN THE PANEL 234 00:10:07,423 --> 00:10:12,228 IN B, OR FIBROTIC PHASE, NOT 235 00:10:12,228 --> 00:10:14,763 SHOWN HERE, THIS DAD IS THE 236 00:10:14,763 --> 00:10:17,032 PREDOMINANT PATHOLOGY IN FATAL 237 00:10:17,032 --> 00:10:19,335 COVID-19 CASES. 238 00:10:19,335 --> 00:10:23,706 BY TAKING OUR CASES, DAVID 239 00:10:23,706 --> 00:10:26,642 KLEINER IN NCI PATHOLOGY LAB WAS 240 00:10:26,642 --> 00:10:29,511 ABLE TO ANNOTATE PRESENCE OF DAD 241 00:10:29,511 --> 00:10:31,380 IN THESE DIFFERENT STAGES ACROSS 242 00:10:31,380 --> 00:10:33,115 EACH OF OUR PATIENTS. 243 00:10:33,115 --> 00:10:36,385 AND THROUGH THAT ANNOTATION WE 244 00:10:36,385 --> 00:10:40,222 DETERMINED THAT BY DAY 14 POST 245 00:10:40,222 --> 00:10:42,324 ILLNESS ONSET, DAD WAS DETECTED 246 00:10:42,324 --> 00:10:45,728 IN THE LUNGS OF ALL OF OUR FATAL 247 00:10:45,728 --> 00:10:48,664 CASES, AND BY ONE MONTH LUNG 248 00:10:48,664 --> 00:10:52,601 FIBROSIS WAS DETECTED IN OVER 249 00:10:52,601 --> 00:10:56,438 50% OF OUR CASES. 250 00:10:56,438 --> 00:11:03,078 WE USED SARS-COV-2 SUBGENOMIC 251 00:11:03,078 --> 00:11:07,049 RNA, A SURROGATE FOR VIRAL 252 00:11:07,049 --> 00:11:09,919 REPLICATION, TO DETERMINE THAT 253 00:11:09,919 --> 00:11:11,453 SARS CoV STOP REPLICATED IN 254 00:11:11,453 --> 00:11:14,556 OUR PATIENTS' LUNG SAMPLES OF 255 00:11:14,556 --> 00:11:17,826 THESE FATAL CASES MEDIAN OF NINE 256 00:11:17,826 --> 00:11:20,095 DAYS AFTER PATIENT RECORDED 257 00:11:20,095 --> 00:11:21,096 ILLNESS ONSET. 258 00:11:21,096 --> 00:11:27,136 DESPITE THE END OF VIRAL 259 00:11:27,136 --> 00:11:28,904 REPLICATION AS NOTED EVERY 260 00:11:28,904 --> 00:11:31,073 PATIENT AFTER DAY 14 DEVELOPED 261 00:11:31,073 --> 00:11:34,109 OR HAVE PROGRESSIVE DAD IN THEIR 262 00:11:34,109 --> 00:11:34,310 LUNGS. 263 00:11:34,310 --> 00:11:35,644 SUGGESTING THAT IT'S NOT JUST 264 00:11:35,644 --> 00:11:37,846 THE VIRUS THAT WE NEED TO TARGET 265 00:11:37,846 --> 00:11:40,482 BUT THAT ACTUALLY WE NEED TO 266 00:11:40,482 --> 00:11:43,018 TARGET THE HOST AND REPAIR 267 00:11:43,018 --> 00:11:47,790 RESPONSES AFTER THE END OF VIRAL 268 00:11:47,790 --> 00:11:54,630 REPLICATION WHEN WE'RE 269 00:11:54,630 --> 00:11:55,931 THERAPEUTICALLY TREATING THEIRS 270 00:11:55,931 --> 00:11:56,699 PATIENTS. 271 00:11:56,699 --> 00:11:59,969 TO BETTER UNDERSTAND WE DESIGNED 272 00:11:59,969 --> 00:12:02,404 AND VALIDATED A GREATER THAN 30 273 00:12:02,404 --> 00:12:05,908 METAL TAGGED ANTIBODY PANEL AND 274 00:12:05,908 --> 00:12:09,311 APPLIED IMAGING MASS CYTOMETRY 275 00:12:09,311 --> 00:12:10,713 TO DEEPLY CHARACTERIZE INNATE 276 00:12:10,713 --> 00:12:15,551 ADAPTIVE IMMUNE CELLS WITHIN 277 00:12:15,551 --> 00:12:17,753 THEIR LUNG SPATIAL CONTEXT. 278 00:12:17,753 --> 00:12:20,456 MYELOID AND PARENCHYMAL CELLS 279 00:12:20,456 --> 00:12:23,726 CAPTURED BY OUR ASSAY INCLUDE 280 00:12:23,726 --> 00:12:25,828 SUBPOPULATIONS OF NEUTROPHILS, 281 00:12:25,828 --> 00:12:31,834 AND MACROPHAGES, AS WELL AS 282 00:12:31,834 --> 00:12:33,168 EPITHELIAL, ENDOTHELIAL, 283 00:12:33,168 --> 00:12:34,036 DENDRITIC CELLS. 284 00:12:34,036 --> 00:12:38,273 WITH THIS PANEL, WE CAN ANALYZE 285 00:12:38,273 --> 00:12:41,910 CHANGES IN CELL POPULATIONS AND 286 00:12:41,910 --> 00:12:43,846 THEIR SPATIAL ORIENTATION 287 00:12:43,846 --> 00:12:46,815 ACCORDING TO OUR WELL 288 00:12:46,815 --> 00:12:48,417 CHARACTERIZED DEMOGRAPHIC 289 00:12:48,417 --> 00:12:50,419 CLINICAL AND VIROLOGIC METADATA 290 00:12:50,419 --> 00:12:51,120 VARIABLES. 291 00:12:51,120 --> 00:12:54,289 IN THE EXAMPLES SHOWN HERE, WE 292 00:12:54,289 --> 00:12:57,226 OBSERVED THAT THE NUMBER OR THE 293 00:12:57,226 --> 00:12:59,962 COUNT OF LUNG MACROPHAGES IN 294 00:12:59,962 --> 00:13:00,596 REGIONS WE EVALUATED, AND THAT 295 00:13:00,596 --> 00:13:03,032 GRAPH IS SHOWN ON THE LEFT, AND 296 00:13:03,032 --> 00:13:04,967 THE NEUTROPHILS, IN THE GRAPH 297 00:13:04,967 --> 00:13:08,404 SHOWN ON THE RIGHT, THAT THOSE 298 00:13:08,404 --> 00:13:09,605 COUNTS DO NOT SIGNIFICANTLY 299 00:13:09,605 --> 00:13:13,442 CHANGE IN THE LUNGS WE EVALUATED 300 00:13:13,442 --> 00:13:15,277 AFTER THE VIRUS STOPS 301 00:13:15,277 --> 00:13:17,479 REPLICATING, AS DETERMINED BY 302 00:13:17,479 --> 00:13:19,648 DETECTION OR NOT OF SUBGENOMIC 303 00:13:19,648 --> 00:13:22,785 RNA IN ASSOCIATED LUNG SAMPLES. 304 00:13:22,785 --> 00:13:24,920 SO THE CONCEPT THERE IS THE 305 00:13:24,920 --> 00:13:26,889 VIRUS STOPS REPLICATING BUT 306 00:13:26,889 --> 00:13:28,757 THESE INNATE IMMUNE CELLS OFTEN 307 00:13:28,757 --> 00:13:31,794 THOUGHT TO CONTRIBUTE TO LUNG 308 00:13:31,794 --> 00:13:32,394 INJURY AND PATHOGENESIS, THEY 309 00:13:32,394 --> 00:13:34,763 PERSIST IN THE LUNGS. 310 00:13:34,763 --> 00:13:38,567 AND I SHOULD NOTE THESE SAMPLES 311 00:13:38,567 --> 00:13:40,669 WERE PARTICULARLY SELECTED TO 312 00:13:40,669 --> 00:13:43,839 HAVE THE ABSENCE OF EVIDENCE OF 313 00:13:43,839 --> 00:13:44,706 BACTERIAL CO-INFECTION IN THEM 314 00:13:44,706 --> 00:13:48,644 WHICH WOULD BE A LOGICAL 315 00:13:48,644 --> 00:13:52,147 RATIONALE TO SAY WHY DO THEY 316 00:13:52,147 --> 00:13:53,582 HAVE ONGOING INNOVATE IMMUNE 317 00:13:53,582 --> 00:13:55,784 CELL RESPONSES IN THESE SAMPLES, 318 00:13:55,784 --> 00:14:00,055 AREAS FREE OF HISTOPATHOLOGIC OR 319 00:14:00,055 --> 00:14:01,423 MOLECULAR EVIDENCE OF 320 00:14:01,423 --> 00:14:01,757 CO-INFECTION. 321 00:14:01,757 --> 00:14:03,892 TO VISUALIZE THE IMPLICATIONS OF 322 00:14:03,892 --> 00:14:07,529 THIS, WE CAN EXAMINE THE LUNG 323 00:14:07,529 --> 00:14:10,899 PARENCHYMA OF A 71-YEAR-OLD 324 00:14:10,899 --> 00:14:13,435 FEMALE WHO DIED 33 DAYS POST 325 00:14:13,435 --> 00:14:14,870 ILLNESS ONSET. 326 00:14:14,870 --> 00:14:17,840 HER LUNG PATHOLOGY AS EXPECTED 327 00:14:17,840 --> 00:14:21,410 SHOWED PROLIFERATIVE AND 328 00:14:21,410 --> 00:14:23,078 FIBROSING DIFFUSE ALVEOLAR 329 00:14:23,078 --> 00:14:25,681 DAMAGE WITH DISORDERED AND 330 00:14:25,681 --> 00:14:26,381 SPARSE ALVEOLAR STRUCTURES, 331 00:14:26,381 --> 00:14:29,451 WHICH ARE INDICATED IN THIS 332 00:14:29,451 --> 00:14:31,753 IMAGE BY EPITHELIAL CELL MARKER 333 00:14:31,753 --> 00:14:35,224 PAN CK, PSEUDOCOLORED MA GENERAL 334 00:14:35,224 --> 00:14:37,326 DA. MAGENTA. 335 00:14:37,326 --> 00:14:39,294 YOU CAN SEE SOME REMNANTS, BUT 336 00:14:39,294 --> 00:14:40,729 VERY FEW, INSTEAD YOU HAVE 337 00:14:40,729 --> 00:14:43,665 ALMOST REMNANTS OF WHAT USED TO 338 00:14:43,665 --> 00:14:46,502 BE AN INTACT ALVEOLI, SCATTERED 339 00:14:46,502 --> 00:14:48,504 THROUGHOUT THIS IMAGE. 340 00:14:48,504 --> 00:14:50,672 WHILE SUBGENOMIC RNA FROM THIS 341 00:14:50,672 --> 00:14:56,178 LUNG SAMPLE IS NEGATIVE, SHE HAD 342 00:14:56,178 --> 00:14:57,146 A PRONOUNCED INTERSTITIAL 343 00:14:57,146 --> 00:14:57,813 MACROPHAGE INFILTRATE INDICATED 344 00:14:57,813 --> 00:15:00,315 IN THIS IMAGE BY CD14 STAINING 345 00:15:00,315 --> 00:15:02,084 WHICH IS PSEUDOCOLORED RED, SO 346 00:15:02,084 --> 00:15:05,120 YOU CAN SEE THIS INTERSTITIAL 347 00:15:05,120 --> 00:15:06,455 MACROPHAGE INFILTRATE, OF COURSE 348 00:15:06,455 --> 00:15:11,126 WE ARE ABLE TO GO ON AND FURTHER 349 00:15:11,126 --> 00:15:11,760 CHARACTERIZE SUBPOPULATIONS OF 350 00:15:11,760 --> 00:15:14,363 MACROPHAGES, I'M NOT SHOWING 351 00:15:14,363 --> 00:15:16,198 THAT HERE. 352 00:15:16,198 --> 00:15:19,134 IN THE SAME IMAGE IN DIFFERENT 353 00:15:19,134 --> 00:15:20,135 SPATIAL ORIENTATION YOU'LL NOTE 354 00:15:20,135 --> 00:15:24,173 THAT THERE ARE CLUSTERS OF 355 00:15:24,173 --> 00:15:26,141 NEUTROPHILS WHICH ARE INDICATED 356 00:15:26,141 --> 00:15:28,577 BY OUR CD15 MARKER, WHICH IS 357 00:15:28,577 --> 00:15:30,345 PSEUDOCOLORED GREEN HERE, BUT 358 00:15:30,345 --> 00:15:31,413 THAT THOSE NEUTROPHILS ARE 359 00:15:31,413 --> 00:15:36,585 PRESENT IN WHAT APPEAR TO BE 360 00:15:36,585 --> 00:15:38,887 ALVEOLAR REMNANTS. 361 00:15:38,887 --> 00:15:41,456 TAKEN TOGETHER, MACROPHAGES AND 362 00:15:41,456 --> 00:15:43,458 NEUTROPHILS ARE DETECTED, 33 363 00:15:43,458 --> 00:15:45,427 DAYS AFTER SYMPTOM ONSET, AND 364 00:15:45,427 --> 00:15:46,228 OCCUPY DISTINCTION SPATIAL 365 00:15:46,228 --> 00:15:48,664 NICHES IN THIS PATIENT'S 366 00:15:48,664 --> 00:15:54,303 SUBGENOMIC RNA AND AS I 367 00:15:54,303 --> 00:15:55,204 MENTIONED BEFORE BACTERIAL 368 00:15:55,204 --> 00:15:59,141 CO-INFECTION FREE, IF YOU WILL, 369 00:15:59,141 --> 00:15:59,942 LUNG SAMPLE. 370 00:15:59,942 --> 00:16:02,644 SUPPORTING THIS IDEA THAT 371 00:16:02,644 --> 00:16:03,612 THERE'S THIS PERSISTENT INNATE 372 00:16:03,612 --> 00:16:05,681 IMMUNE CELL IN THESE FATAL CASES 373 00:16:05,681 --> 00:16:08,884 IN THE LUNGS. 374 00:16:08,884 --> 00:16:12,154 NOW, AS DESIGNED OR IMC PANEL 375 00:16:12,154 --> 00:16:14,790 ALSO ALLOWS US TO CHARACTERIZE 376 00:16:14,790 --> 00:16:17,292 T- AND B-CELL POPULATIONS 377 00:16:17,292 --> 00:16:19,361 INCLUDING NAIVE AND ANTIGENIC 378 00:16:19,361 --> 00:16:21,163 EXPERIENCE, MEMORY OR EFFECTOR 379 00:16:21,163 --> 00:16:24,433 CD4 AND CD8 T CELLS, THAT'S 380 00:16:24,433 --> 00:16:31,306 REPRESENTED BY THIS BIORENDER 381 00:16:31,306 --> 00:16:33,075 IMAGE HERE. 382 00:16:33,075 --> 00:16:35,377 IN CONTRAST, CD4 T CELLS SHOWN 383 00:16:35,377 --> 00:16:38,347 ON THE GRAPH ON THE LEFT, CD8 T 384 00:16:38,347 --> 00:16:40,515 CELLS SHOWN ON THE GRAPH IN THE 385 00:16:40,515 --> 00:16:42,851 MIDDLE, AND B CELLS SHOWN ON THE 386 00:16:42,851 --> 00:16:46,021 GRAPH ON THE RIGHT, THOSE 387 00:16:46,021 --> 00:16:49,057 POPULATIONS AND ALL THREE 388 00:16:49,057 --> 00:16:51,593 INCREASE IN OUR LUNGS AFTER 389 00:16:51,593 --> 00:16:53,328 SARS-COV-2 STOPS REPLICATING. 390 00:16:53,328 --> 00:16:54,763 AND THAT'S, AGAIN, DETERMINED BY 391 00:16:54,763 --> 00:16:57,065 PRESENCE OR ABSENCE OF OUR 392 00:16:57,065 --> 00:16:59,034 SUBGENOMIC RNA AND OUR 393 00:16:59,034 --> 00:17:02,738 ASSOCIATED LUNG SAMPLES. 394 00:17:02,738 --> 00:17:04,940 THERE'S A BOOST IN IMMUNE 395 00:17:04,940 --> 00:17:07,042 ADAPTIVE RESPONSES EVEN AFTER 396 00:17:07,042 --> 00:17:08,243 THE VIRUS STOPS REPLICATING. 397 00:17:08,243 --> 00:17:11,847 NOW, IF WE GO BACK TO OUR LUNG 398 00:17:11,847 --> 00:17:13,482 OF OUR 71-YEAR-OLD FEMALE 399 00:17:13,482 --> 00:17:16,685 PATIENT WHO DIED 33 DAYS POST 400 00:17:16,685 --> 00:17:19,388 ILLNESS ONSET, WE OBSERVE AND 401 00:17:19,388 --> 00:17:20,589 INTERSTITIAL T CELL INFILTRATE. 402 00:17:20,589 --> 00:17:23,892 NOW, THAT IS SHOWN BY OUR CD3 403 00:17:23,892 --> 00:17:25,460 STAINING WHICH IS BLUE, AND IN A 404 00:17:25,460 --> 00:17:28,497 WAY THE DISTRIBUTION OF THOSE T 405 00:17:28,497 --> 00:17:30,666 CELLS SOMEWHAT MATCHES THE 406 00:17:30,666 --> 00:17:31,767 DISTRIBUTION OF OUR MACROPHAGES, 407 00:17:31,767 --> 00:17:33,935 AND OF COURSE ACROSS ALL OF OUR 408 00:17:33,935 --> 00:17:35,604 PATIENTS AND ACROSS ALL OF OUR 409 00:17:35,604 --> 00:17:38,006 TIME WE'LL BE ABLE TO 410 00:17:38,006 --> 00:17:39,207 CHARACTERIZE THE SPATIAL 411 00:17:39,207 --> 00:17:43,812 ORIENTATION OF DIFFERENT CELL 412 00:17:43,812 --> 00:17:44,880 SUBSETS. 413 00:17:44,880 --> 00:17:47,215 WHILE WE HAVE MANY INFILTRATING 414 00:17:47,215 --> 00:17:51,486 T CELLS, WE HAVE MANY FEWER 415 00:17:51,486 --> 00:17:53,355 INTERSTITIAL B CELLS INDICATED 416 00:17:53,355 --> 00:17:58,093 BY OUR CD20 STAIN AND THAT'S 417 00:17:58,093 --> 00:18:00,262 CYAN. 418 00:18:00,262 --> 00:18:01,596 YOU HAVE 419 00:18:01,596 --> 00:18:02,898 YOU HAVE TO HUNT BUT THEY ARE 420 00:18:02,898 --> 00:18:03,332 THERE. 421 00:18:03,332 --> 00:18:05,734 THE WORK IN PROGRESS, THIS 422 00:18:05,734 --> 00:18:07,035 SUMMARY, OVERALL AIMS TO PROVIDE 423 00:18:07,035 --> 00:18:09,671 BETTER UNDERSTAND OF HOW INNATE 424 00:18:09,671 --> 00:18:10,972 AND ADAPTIVE IMMUNE CELLS 425 00:18:10,972 --> 00:18:12,741 INTERACT WITH EACH OTHER AND 426 00:18:12,741 --> 00:18:15,711 OTHER PARENCHYMAL CELLS WE CAN 427 00:18:15,711 --> 00:18:19,548 CAPTURE EPITHELIAL AND WE CAN 428 00:18:19,548 --> 00:18:21,983 CAPTURE ENDOTHELIAL CELLS IN THE 429 00:18:21,983 --> 00:18:22,951 LUNG PARENCHYMA DURING FATAL 430 00:18:22,951 --> 00:18:25,687 COVID-19, THE HOPE IS THIS WILL 431 00:18:25,687 --> 00:18:28,490 PROVIDE US INSIGHTS PERHAPS INTO 432 00:18:28,490 --> 00:18:30,926 WHICH HOST RESPONSES MIGHT BE 433 00:18:30,926 --> 00:18:31,927 THERAPEUTICALLY TARGETED IN 434 00:18:31,927 --> 00:18:34,896 THESE SEVERE AND FATAL CASES, 435 00:18:34,896 --> 00:18:37,265 AND AT WHICH POINT DURING 436 00:18:37,265 --> 00:18:42,104 ILLNESS FROM EARLY TO LATE 437 00:18:42,104 --> 00:18:43,038 DISEASE PROGRESSION. 438 00:18:43,038 --> 00:18:48,343 SO, TO GAIN A BETTER INSIGHT 439 00:18:48,343 --> 00:18:48,977 INTO SARS-COV-2 SPECIFIC T-CELL 440 00:18:48,977 --> 00:18:51,279 RESPONSES IN THE LUNGS OF OUR 441 00:18:51,279 --> 00:18:54,249 FATAL COVID-19 CASES, WE 442 00:18:54,249 --> 00:18:55,250 PERFORMED T-CELL RECEPTOR BETA 443 00:18:55,250 --> 00:18:58,420 SEQUENCE OF SEQUENCING OF LUNG 444 00:18:58,420 --> 00:19:01,690 TISSUES ACROSS OUR COHORT. 445 00:19:01,690 --> 00:19:05,293 V(D)J REARRANGEMENT OF THE CDR 3 446 00:19:05,293 --> 00:19:11,500 VARIABLE REGION OF THE CHAIN 447 00:19:11,500 --> 00:19:14,136 DETERMINES T CELL ANTIGEN 448 00:19:14,136 --> 00:19:14,703 SPECIFICITY. 449 00:19:14,703 --> 00:19:15,370 ADAPTIVE BIOTECHNOLOGIES WHICH 450 00:19:15,370 --> 00:19:19,975 IS THE TOOL WE USED TO DO THE 451 00:19:19,975 --> 00:19:22,744 SEQUENCING USES A PROPRIETARY 452 00:19:22,744 --> 00:19:24,312 FUNCTIONAL MULTIPLEX ASSAY FOR 453 00:19:24,312 --> 00:19:27,783 THE IDENTIFICATION OF T-CELL 454 00:19:27,783 --> 00:19:28,550 RECEPTOR ANTIGEN SPECIFICITY. 455 00:19:28,550 --> 00:19:30,252 THEY'VE DONE A NUMBER OF 456 00:19:30,252 --> 00:19:32,053 FUNCTIONAL STUDIES. 457 00:19:32,053 --> 00:19:36,858 THEY HAVE DETERMINED THE 458 00:19:36,858 --> 00:19:38,293 DISTRIBUTION OF 459 00:19:38,293 --> 00:19:39,261 SARS-COV-2-SPECIFIC CLONOTYPES, 460 00:19:39,261 --> 00:19:41,430 AND THEIR COGNATE ANTIGENS AND 461 00:19:41,430 --> 00:19:42,964 WITH THAT FUNCTIONAL DATASET 462 00:19:42,964 --> 00:19:45,066 THAT IS A PUBLICLY AVAILABLE 463 00:19:45,066 --> 00:19:46,802 DATASET, COULD BE USED AS A 464 00:19:46,802 --> 00:19:49,638 RESOURCE FOR US TO COMPARE OUR 465 00:19:49,638 --> 00:19:53,175 T-CELL RECEPTOR CLONOTYPES, AND 466 00:19:53,175 --> 00:19:57,245 TO UNDERSTAND THE BREADTH, THE 467 00:19:57,245 --> 00:20:00,182 DEPTH, ANTIGEN SPECIFICITY AND 468 00:20:00,182 --> 00:20:02,350 SPECIFIC COV 2 ANTIGENS. 469 00:20:02,350 --> 00:20:04,352 WITH THIS RESOURCE, AS A 470 00:20:04,352 --> 00:20:07,088 COMPONENT OF A LARGER, AGAIN, 471 00:20:07,088 --> 00:20:09,691 ONGOING PROJECT, WE OBSERVED 472 00:20:09,691 --> 00:20:15,163 THAT THE RATIO OF SARS-COV-2 473 00:20:15,163 --> 00:20:17,466 SPECIFIC CLONE TO NON-SPECIFIC 474 00:20:17,466 --> 00:20:19,000 CLONES DOES NOT SIGNIFICANTLY 475 00:20:19,000 --> 00:20:21,169 CHANGE AS A FUNCTION OF 476 00:20:21,169 --> 00:20:22,404 SUBGENOMIC RNA POSITIVE OR 477 00:20:22,404 --> 00:20:23,772 NEGATIVE STATUS, AND THAT'S THE 478 00:20:23,772 --> 00:20:26,241 GRAPH SHOWN ON THE LEFT. 479 00:20:26,241 --> 00:20:29,444 NOW, CLONES, YOU CAN THINK OF 480 00:20:29,444 --> 00:20:31,480 THESE AS INDIVIDUAL T CELLS, 481 00:20:31,480 --> 00:20:33,949 SAYING ALL YOUR T CELLS THAT ARE 482 00:20:33,949 --> 00:20:35,851 SPECIFIC TO COV 2 OVER ALL OF 483 00:20:35,851 --> 00:20:37,919 THE T CELLS PRESENT IN THE LUNG, 484 00:20:37,919 --> 00:20:39,888 THAT'S THE RATIO, DID NOT 485 00:20:39,888 --> 00:20:41,323 MEANINGFULLY CHANGE BEFORE AND 486 00:20:41,323 --> 00:20:45,260 AFTER VIRUS STOPPED REPLICATING. 487 00:20:45,260 --> 00:20:49,431 SIMILARLY THE SAME WAS OBSERVED 488 00:20:49,431 --> 00:20:50,899 FOR SARS-COV-2 SPECIFIC 489 00:20:50,899 --> 00:20:51,199 CLONOTYPES. 490 00:20:51,199 --> 00:20:54,569 NOW A CLONOTYPE HAS AN 491 00:20:54,569 --> 00:20:56,204 ANTIGEN-SPECIFIC TARGET, ONE 492 00:20:56,204 --> 00:20:57,439 CLONOTYPE CAN HAVE MULTIPLE 493 00:20:57,439 --> 00:20:59,708 CLONES EXPANDED. 494 00:20:59,708 --> 00:21:02,210 BUT THE NUMBER OF CLONOTYPES 495 00:21:02,210 --> 00:21:04,212 TELLS YOU BREADTH OF YOUR T-CELL 496 00:21:04,212 --> 00:21:06,381 RESPONSE AGAINST ANTIGENS OF 497 00:21:06,381 --> 00:21:06,648 INTEREST. 498 00:21:06,648 --> 00:21:09,251 AND THE CLONOTYPE DATA ON THE 499 00:21:09,251 --> 00:21:11,319 RIGHT TELLS US THAT THE BREADTH 500 00:21:11,319 --> 00:21:15,023 OF THAT CLONOTYPE, THE RATIO OF 501 00:21:15,023 --> 00:21:16,358 SARS-COV-2 OVER TOTAL DOESN'T 502 00:21:16,358 --> 00:21:17,459 MEANINGFULLY CHANGE BEFORE AND 503 00:21:17,459 --> 00:21:17,959 AFTER. 504 00:21:17,959 --> 00:21:19,828 NOW, THERE'S A NUMBER OF WAYS TO 505 00:21:19,828 --> 00:21:20,829 INTERPRET THIS DATA. 506 00:21:20,829 --> 00:21:23,665 WE'RE STILL WORKING THROUGH IT. 507 00:21:23,665 --> 00:21:25,867 BUT ONE WAY TO INTERPRET IT IS 508 00:21:25,867 --> 00:21:29,905 THAT EVEN AFTER VIRUS STOPS 509 00:21:29,905 --> 00:21:31,006 REPLICATING, WE'VE ALREADY SHOWN 510 00:21:31,006 --> 00:21:33,542 VIRAL RNA PERSISTS IN THE LUNGS, 511 00:21:33,542 --> 00:21:36,578 THAT THERE'S A PERSISTENT AND 512 00:21:36,578 --> 00:21:38,547 DIVERSE T-CELL RESPONSE THAT'S 513 00:21:38,547 --> 00:21:40,215 HAPPENING WITHIN THESE LUNGS AND 514 00:21:40,215 --> 00:21:41,950 IN OUR COHORT THAT'S UP TO SEVEN 515 00:21:41,950 --> 00:21:46,888 MONTHS AFTER INITIAL ILLNESS 516 00:21:46,888 --> 00:21:47,622 ONSET. 517 00:21:47,622 --> 00:21:49,291 NOW, MOVING OUT OF THE LUNGS, 518 00:21:49,291 --> 00:21:53,094 AND INTO THE REST OF THE BODY, 519 00:21:53,094 --> 00:21:55,830 WE DETECT VIRAL RNA IN ALL MAJOR 520 00:21:55,830 --> 00:22:00,769 ORGAN SYSTEMS WITH THE HIGHEST 521 00:22:00,769 --> 00:22:05,574 BURDEN DETECTED EARLY AFTER 522 00:22:05,574 --> 00:22:13,915 ILLNESS ONSET WITH VARIABLE 523 00:22:13,915 --> 00:22:17,185 PERSISTENCE OF VIRAL RNA, 524 00:22:17,185 --> 00:22:18,186 ADDITIONALLY GREW VIRUS, 525 00:22:18,186 --> 00:22:24,559 REPLICATION COMPETENT VIRUS FROM 526 00:22:24,559 --> 00:22:26,061 SITES INCLUDING JEJENUM, HEART, 527 00:22:26,061 --> 00:22:27,829 RENAL GLAND, THE EYE. 528 00:22:27,829 --> 00:22:30,365 THERE'S GENERAL CONSENSUS IN THE 529 00:22:30,365 --> 00:22:31,766 FIELD THAT, AGAIN, THERE'S STILL 530 00:22:31,766 --> 00:22:34,302 SOME DEBATE BUT I THINK NOW 531 00:22:34,302 --> 00:22:37,772 THERE'S GENERAL CONSENSUS IN THE 532 00:22:37,772 --> 00:22:39,674 FIELD THAT SARS CoV HAS 533 00:22:39,674 --> 00:22:41,743 ABILITY TO INFECT TISSUES 534 00:22:41,743 --> 00:22:43,178 THROUGHOUT THE BODY, THERE'S 535 00:22:43,178 --> 00:22:44,813 LESS CLARITY ON HOW THE VIRUS 536 00:22:44,813 --> 00:22:48,850 DISSEMINATES TO ALL OF THESE 537 00:22:48,850 --> 00:22:49,050 SITES. 538 00:22:49,050 --> 00:22:51,286 OUR DATA IN OUR COHORT WOULD 539 00:22:51,286 --> 00:22:54,889 SUGGEST THAT THE VIRUS TRAFFICS 540 00:22:54,889 --> 00:22:56,625 IN THE NON-CELLULAR FRACTION OF 541 00:22:56,625 --> 00:23:00,128 THE BLOOD EARLY IN INFECTION AND 542 00:23:00,128 --> 00:23:05,700 PRIOR TO THE ONSET OF HUMORAL 543 00:23:05,700 --> 00:23:06,001 IMMUNITIES. 544 00:23:06,001 --> 00:23:08,436 WE DETECTED VIRAL RNA IN THE 545 00:23:08,436 --> 00:23:12,607 PLASMA OF 11 OF 16, OR 69% OF 546 00:23:12,607 --> 00:23:14,976 CASES WHO DIED WITHIN 14 DAYS OF 547 00:23:14,976 --> 00:23:15,744 ILLNESS ONSET, THAT'S THE DATA 548 00:23:15,744 --> 00:23:17,178 SHOWN ON TOP. 549 00:23:17,178 --> 00:23:19,147 THE HIGHEST BURDEN OF THAT VIRAL 550 00:23:19,147 --> 00:23:23,685 RNA IN THE PLASMA WAS DETECTED 551 00:23:23,685 --> 00:23:25,820 IN EARLY CASES. 552 00:23:25,820 --> 00:23:31,192 WE QUERIED THE PBMC FRACTION FOR 553 00:23:31,192 --> 00:23:33,595 VIRAL RNA IN CASES WITH AND 554 00:23:33,595 --> 00:23:35,797 WITHOUT VIRAL RNA DETECTED IN 555 00:23:35,797 --> 00:23:39,100 THE PLASMA AND DETECTED ALMOST 556 00:23:39,100 --> 00:23:42,537 NO VIRAL RNA IN THE CELLULAR 557 00:23:42,537 --> 00:23:42,804 FRACTION. 558 00:23:42,804 --> 00:23:43,872 AGAIN, SUGGESTING THAT THIS IS 559 00:23:43,872 --> 00:23:45,940 VIRUS CIRCULATING IN THE PLASMA 560 00:23:45,940 --> 00:23:48,910 AND NOT THE CELLULAR FRACTION. 561 00:23:48,910 --> 00:23:51,646 WITHIN THIS GROUP OF PATIENTS, 562 00:23:51,646 --> 00:23:53,481 PATIENT 27 STUCK OUT. 563 00:23:53,481 --> 00:23:55,684 THEY STUCK OUT AS BEING THE 564 00:23:55,684 --> 00:23:56,918 EARLIEST PATIENT THAT WAS 565 00:23:56,918 --> 00:23:59,187 PRESENT IN OUR COHORT WHO SHOWED 566 00:23:59,187 --> 00:24:01,623 UP AFTER ILLNESS ONSET AS HAVING 567 00:24:01,623 --> 00:24:03,925 THE HIGHEST BURDEN OF VIRAL RNA 568 00:24:03,925 --> 00:24:05,794 IN THEIR PLASMA. 569 00:24:05,794 --> 00:24:08,196 AND IS BEING ONLY ONE OF THREE 570 00:24:08,196 --> 00:24:10,699 PATIENTS IN OUR COMPLETE COHORT, 571 00:24:10,699 --> 00:24:15,537 ALL 44 PATIENTS, WHO WAS FULLY 572 00:24:15,537 --> 00:24:17,739 SERONEGATIVE AGAINST SARS-COV-2 573 00:24:17,739 --> 00:24:20,041 NP AND SPIKE PROTEINS. 574 00:24:20,041 --> 00:24:22,777 IN THIS PATIENT, WE SUCCESSFULLY 575 00:24:22,777 --> 00:24:24,512 GREW VIRUS FROM THEIR PLASMA. 576 00:24:24,512 --> 00:24:26,815 AND WE DETERMINED THAT THAT 577 00:24:26,815 --> 00:24:28,683 VIRUS, THE NEAR FULL LENGTH 578 00:24:28,683 --> 00:24:31,086 SEQUENCE OF THAT VIRUS, HAD 579 00:24:31,086 --> 00:24:33,588 ESSENTIALLY THE EXACT SAME 580 00:24:33,588 --> 00:24:36,424 SEQUENCE AS VIRUS THAT WE 581 00:24:36,424 --> 00:24:40,729 EXTRACTED RNA FROM LUNG AND 582 00:24:40,729 --> 00:24:40,929 HEART. 583 00:24:40,929 --> 00:24:44,766 THIS CASE PROVIDES PROOF OF 584 00:24:44,766 --> 00:24:45,700 PRINCIPLE THAT REPLICATION 585 00:24:45,700 --> 00:24:47,602 SARS-COV-2 CAN TRAFFIC IN THE 586 00:24:47,602 --> 00:24:54,209 BLOOD IN SOME CASES AND CAN SEED 587 00:24:54,209 --> 00:24:55,410 DISTANT TISSUES. 588 00:24:55,410 --> 00:24:57,345 ONE QUESTION THAT NATURALLY 589 00:24:57,345 --> 00:24:59,013 ARISES RELATED TO DETECTION OF 590 00:24:59,013 --> 00:25:00,248 VIRUS IN NON-RESPIRATORY TISSUES 591 00:25:00,248 --> 00:25:02,283 IS POTENTIAL FOR BLOOD TO 592 00:25:02,283 --> 00:25:04,252 CONTAMINATE THOSE TISSUES. 593 00:25:04,252 --> 00:25:05,987 RATHER THAN THE TISSUES 594 00:25:05,987 --> 00:25:07,956 THEMSELVES BEING INFECTED. 595 00:25:07,956 --> 00:25:10,692 TO ADDRESS THIS QUESTION AND 596 00:25:10,692 --> 00:25:14,796 VALIDATE OUR PCR FINDINGS, WE 597 00:25:14,796 --> 00:25:18,500 ALSO UTILIZED IN SITU 598 00:25:18,500 --> 00:25:19,701 HYBRIDIZATION ASSAY PROBING 599 00:25:19,701 --> 00:25:21,469 SPIKE GENE TO CONFIRM INFECTION 600 00:25:21,469 --> 00:25:23,204 OF TISSUES AND DETERMINE CELL 601 00:25:23,204 --> 00:25:24,072 TYPE SPECIFICITY OF INFECTION 602 00:25:24,072 --> 00:25:27,041 WITHIN THOSE TISSUES. 603 00:25:27,041 --> 00:25:30,111 IN TOTAL, WE DETECTED SARS-COV-2 604 00:25:30,111 --> 00:25:35,817 RNA WITHIN GREATER THAN 30 CELL 605 00:25:35,817 --> 00:25:39,788 TYPES ACROSS 35 TISSUES. 606 00:25:39,788 --> 00:25:42,524 NOTABLY, IN ALL TISSUES OUTSIDE 607 00:25:42,524 --> 00:25:45,994 OF THE RESPIRATORY TRACT, WE 608 00:25:45,994 --> 00:25:49,164 DETECTED A PAUCITY OF 609 00:25:49,164 --> 00:25:52,367 INFLAMMATORY OR CYTO PATHOLOGY, 610 00:25:52,367 --> 00:25:54,335 INFLAMMATION OR CYTO PATHOLOGY 611 00:25:54,335 --> 00:25:56,638 DIRECTLY ATTRIBUTABLE TO VIRAL 612 00:25:56,638 --> 00:25:56,905 INEFFECT. 613 00:25:56,905 --> 00:25:58,173 ASIDE FROM THE RESPIRATORY TRACT 614 00:25:58,173 --> 00:26:01,543 IN MAJORITY IF NOT ALL OF OUR 615 00:26:01,543 --> 00:26:05,280 TISSUES WE NOTED AN ABSENCE OF 616 00:26:05,280 --> 00:26:07,115 AN INFLAMMATORY CELL INFILTRATE 617 00:26:07,115 --> 00:26:14,255 AND NOTED ABSENCE OF VIRALLY 618 00:26:14,255 --> 00:26:15,223 ATTRIBUTED CYTOPATHOLOGY. 619 00:26:15,223 --> 00:26:20,929 ALSO IN THE CASE IN EYE, A STUDY 620 00:26:20,929 --> 00:26:24,265 WITH THE NEI, DETECTED 621 00:26:24,265 --> 00:26:29,971 SARS-COV-2 SPIKE RNA IN 622 00:26:29,971 --> 00:26:31,472 SCLEROFIBROBLASTS IN PANEL A, 623 00:26:31,472 --> 00:26:32,774 RETINAL OUTER AND INNER NUCLEAR 624 00:26:32,774 --> 00:26:38,580 LAYERS SHOWN IN PANEL C, AND 625 00:26:38,580 --> 00:26:46,354 CORNEAL EPITHELIUM IN PANEL D 626 00:26:46,354 --> 00:26:48,857 WITHOUT CYTO PATHOLOGY. 627 00:26:48,857 --> 00:26:53,895 AND FINALLY, MOVING ON TO THE 628 00:26:53,895 --> 00:26:59,067 BRAIN, WE DETECTED SARS-COV-2 629 00:26:59,067 --> 00:27:01,369 RNA IN MULTIPLE NON-CONTIGUOUS 630 00:27:01,369 --> 00:27:04,405 REGIONS IN 10 OF 11 BRAINS THAT 631 00:27:04,405 --> 00:27:07,709 WE EVALUATED. 632 00:27:07,709 --> 00:27:13,615 THIS NON-CONTIGUOUS DISTRIBUTION 633 00:27:13,615 --> 00:27:15,783 SUGGESTS HEMO TOGENOUS SEEDING 634 00:27:15,783 --> 00:27:17,318 RATHER THAN RETROGRADE SPREAD 635 00:27:17,318 --> 00:27:18,519 VIA OLFACTORY TRACT ALONE, NOT 636 00:27:18,519 --> 00:27:20,188 SAYING ONE HAPPENS AND THE OTHER 637 00:27:20,188 --> 00:27:22,290 DOESN'T, THEY ARE NOT MUTUALLY 638 00:27:22,290 --> 00:27:24,893 EXCLUSIONIVE, THEY CAN BOTH 639 00:27:24,893 --> 00:27:25,460 HAPPEN. 640 00:27:25,460 --> 00:27:28,596 BUT OUR DATA, BASED UPON THE 641 00:27:28,596 --> 00:27:29,497 DISTRIBUTION, SUGGESTS THAT 642 00:27:29,497 --> 00:27:35,370 INFECTION IN THESE INDIVIDUALS 643 00:27:35,370 --> 00:27:36,604 WAS ATTRIBUTABLE TO HEMOTOGENOUS 644 00:27:36,604 --> 00:27:36,838 SEEDING. 645 00:27:36,838 --> 00:27:45,580 FROM PATIENT 38, THIRD FROM THE 646 00:27:45,580 --> 00:27:47,482 LEFT, THE HASH MARKS ARE 647 00:27:47,482 --> 00:27:49,851 SUBGENOMIC RNA, THAT WAS OUR 648 00:27:49,851 --> 00:27:52,020 MOLECULAR MARKER OF REPLICATION, 649 00:27:52,020 --> 00:27:53,554 SUGGESTING THERE WAS A LOT OF 650 00:27:53,554 --> 00:27:55,623 RNA AND THERE WAS A SUGGESTION 651 00:27:55,623 --> 00:27:56,791 THAT VIRUS WAS REPLICATING IN 652 00:27:56,791 --> 00:27:59,127 THOSE TISSUES. 653 00:27:59,127 --> 00:28:03,631 IT WAS WITHIN THAT TISSUE THAT 654 00:28:03,631 --> 00:28:05,433 WE SUCCESSFULLY ISOLATED 655 00:28:05,433 --> 00:28:09,137 SARS-COV-2 FROM THE BRAIN IN 656 00:28:09,137 --> 00:28:11,105 PATIENT 38, PROVING THE 657 00:28:11,105 --> 00:28:14,375 POTENTIAL FOR VIABLE SARS-COV-2 658 00:28:14,375 --> 00:28:16,778 INFECTION OF THE BRAIN. 659 00:28:16,778 --> 00:28:20,148 FURTHER, WE CONFIRMED SARS-COV-2 660 00:28:20,148 --> 00:28:22,483 INFECTION OF NEURONS IN BRAIN 661 00:28:22,483 --> 00:28:30,358 AND SPINAL CORD VIA FLUORESCENT 662 00:28:30,358 --> 00:28:40,835 HEMO CHEMISTRY, VALIDATED, 663 00:28:42,136 --> 00:28:43,171 CONFIRMING THESE, THOSE IMAGES 664 00:28:43,171 --> 00:28:46,341 I'M NOT SHOWING YOU. 665 00:28:46,341 --> 00:28:48,309 AND CONSISTENT WITH VIRAL 666 00:28:48,309 --> 00:28:52,146 REPLICATION IN THE BRAIN, AND 667 00:28:52,146 --> 00:28:54,649 INDICATIVE OF VIRAL EVOLUTION IN 668 00:28:54,649 --> 00:28:59,721 THE BRAIN, WE DETECTED UNIQUE 669 00:28:59,721 --> 00:29:00,788 SARS-COV-2 SPIKE GENE VARIANTS, 670 00:29:00,788 --> 00:29:02,457 THIS IS LONG READ SEQUENCING 671 00:29:02,457 --> 00:29:05,059 THAT WAS DONE BY ELY AND HIS 672 00:29:05,059 --> 00:29:09,197 GROUP IN NIAID, THAT WE DETECTED 673 00:29:09,197 --> 00:29:10,999 THESE UNIQUE SARS-COV-2 SPIKE 674 00:29:10,999 --> 00:29:15,169 VARIANTS IN BRAIN TISSUE 675 00:29:15,169 --> 00:29:15,903 SPECIFICALLY THALAMUS AND HYPO 676 00:29:15,903 --> 00:29:19,307 THAT 677 00:29:19,307 --> 00:29:19,841 HYPOTHALAMUS, PATIENT 38, 678 00:29:19,841 --> 00:29:21,943 PATIENT WITH THE HOT BRAIN. 679 00:29:21,943 --> 00:29:22,944 THOSE SEQUENCES, THOSE VARIANTS 680 00:29:22,944 --> 00:29:25,446 IN THE BRAIN, WERE NOT DETECTED 681 00:29:25,446 --> 00:29:28,950 IN THIS PATIENT'S LUNG SAMPLES, 682 00:29:28,950 --> 00:29:30,284 SUGGESTING REPLICATION OR 683 00:29:30,284 --> 00:29:33,888 PERHAPS VIRAL EVOLUTION IN 684 00:29:33,888 --> 00:29:36,724 DIFFERENT ANATOMIC SITES. 685 00:29:36,724 --> 00:29:37,925 DESPITE DETECTION OF VIRAL RNA 686 00:29:37,925 --> 00:29:40,328 IN MULTIPLE BRAIN TISSUES WE 687 00:29:40,328 --> 00:29:43,064 AGAIN OBSERVED A PAUCITY OF 688 00:29:43,064 --> 00:29:44,932 INFLAMMATION THAT COULD BE 689 00:29:44,932 --> 00:29:52,040 ATTRIBUTED TO THE - OR CYTO 690 00:29:52,040 --> 00:29:55,543 PATHOLOGY ATTRIBUTED TO THE 691 00:29:55,543 --> 00:29:56,177 VIRUS. 692 00:29:56,177 --> 00:29:57,311 WE HYPOTHESIZE NEURONAL CHANGE 693 00:29:57,311 --> 00:30:00,481 MIGHT BE OCCURRING AT 694 00:30:00,481 --> 00:30:01,783 TRANSCRIPTIONAL LEVEL WITH 695 00:30:01,783 --> 00:30:02,450 POTENTIAL PHENOTYPIC 696 00:30:02,450 --> 00:30:03,184 CONSEQUENCES OR FUNCTIONAL 697 00:30:03,184 --> 00:30:05,620 CONSEQUENCES, TO EVALUATE WE 698 00:30:05,620 --> 00:30:09,023 SELECTED CEREBELLUM FOR A PILOT 699 00:30:09,023 --> 00:30:13,428 STUDY USING SPATIAL 700 00:30:13,428 --> 00:30:17,432 TRANSCRIPTOMICS TO QUERY 701 00:30:17,432 --> 00:30:18,433 DIFFERENTIAL GENE EXPRESSION IN 702 00:30:18,433 --> 00:30:20,735 CEREBELLAR TISSUE. 703 00:30:20,735 --> 00:30:22,904 WE SELECTED CEREBELLUM FOR PILOT 704 00:30:22,904 --> 00:30:25,973 STUDY BECAUSE OF DISTINCT 705 00:30:25,973 --> 00:30:29,043 ANATOMY OF THE CEREBELLUM, THE 706 00:30:29,043 --> 00:30:32,880 FOLIA SHOWN HERE, THAT HAS THIS 707 00:30:32,880 --> 00:30:36,050 NEURON DENSE GRANULAR LAYER, 708 00:30:36,050 --> 00:30:39,654 ADJACENT OUTER MOLECULAR LAYER, 709 00:30:39,654 --> 00:30:45,026 CENTRAL WHITE MATTER LAYER. 710 00:30:45,026 --> 00:30:49,063 WE USED 10X VISIUM PLATFORM UP 711 00:30:49,063 --> 00:30:53,868 TO 14,000 SPOTS ON THIS 11 X 712 00:30:53,868 --> 00:30:55,403 11-MILLIMETER SLIDE. 713 00:30:55,403 --> 00:30:57,805 GIVEN AN APPROXIMATE NEURON CELL 714 00:30:57,805 --> 00:31:00,007 BODY DIAMETER OF ABOUT FIVE 715 00:31:00,007 --> 00:31:01,809 MICRONS, EACH OF THE SPOTS 716 00:31:01,809 --> 00:31:04,812 REPRESENTS THE GENE EXPRESSION 717 00:31:04,812 --> 00:31:06,814 PROFILE OF AROUND 10 NEURONS 718 00:31:06,814 --> 00:31:08,983 WITHIN THE GRANULAR LAYER. 719 00:31:08,983 --> 00:31:11,819 IN THIS EXAMPLE UNBIASED 720 00:31:11,819 --> 00:31:15,223 CLUSTERING OF PATIENT 41 721 00:31:15,223 --> 00:31:16,023 CEREBELLUM SHOWS DOMINANT 722 00:31:16,023 --> 00:31:17,325 CLUSTERS ACCORDING TO GRANULAR 723 00:31:17,325 --> 00:31:20,962 MOLECULAR AND WHITE MATTER 724 00:31:20,962 --> 00:31:22,063 LAYERS, WITH MULTIPLE ADDITIONAL 725 00:31:22,063 --> 00:31:26,634 LESS PROMINENT CLUSTERS THAT 726 00:31:26,634 --> 00:31:27,535 WERE IDENTIFIED. 727 00:31:27,535 --> 00:31:32,540 DECON CONVOLUTION OF CELL TYPES 728 00:31:32,540 --> 00:31:35,276 WITHIN SPOTS FROM EACH LAYER 729 00:31:35,276 --> 00:31:37,745 REVEALS THAT AS EXPECTED 730 00:31:37,745 --> 00:31:38,880 EXCITATORY NEURONS PREDOMINATE 731 00:31:38,880 --> 00:31:45,253 IN THE GRANULAR LAYER OF THE 732 00:31:45,253 --> 00:31:45,686 CEREBELLUM. 733 00:31:45,686 --> 00:31:49,056 FOCUSING ONLY ON THE EXCITATORY 734 00:31:49,056 --> 00:31:50,825 NEURON DENSE CEREBELLAR GRANULAR 735 00:31:50,825 --> 00:31:54,662 LAYER, WE WERE ABLE TO MANUALLY 736 00:31:54,662 --> 00:31:57,832 ANNOTATE THE BURDEN OF 737 00:31:57,832 --> 00:32:01,169 SARS-COV-2 INFECTION ACROSS OUR 738 00:32:01,169 --> 00:32:05,706 PATIENTS ON ADJACENT 5 MICRO 739 00:32:05,706 --> 00:32:07,141 METER SLICE OF TISSUE AS 740 00:32:07,141 --> 00:32:09,177 DETERMINED BY ISH. 741 00:32:09,177 --> 00:32:11,746 SOME PATIENTS INCLUDING 41 AND 742 00:32:11,746 --> 00:32:15,216 42 HAD HIGH BURDEN OF INFECTION 743 00:32:15,216 --> 00:32:16,017 IN CEREBELLAR GRANULAR LAYER, 744 00:32:16,017 --> 00:32:19,720 OTHERS BURDEN OF INFECTION WAS 745 00:32:19,720 --> 00:32:21,589 QUITE LOW. 746 00:32:21,589 --> 00:32:24,926 WHILE WE WERE UNDERPOWERED TO 747 00:32:24,926 --> 00:32:26,827 DETECT SIGNIFICANT GENE 748 00:32:26,827 --> 00:32:29,697 EXPRESSION DIFFERENCES IN 749 00:32:29,697 --> 00:32:32,400 INFECTED VERSUS UNINFECTED 750 00:32:32,400 --> 00:32:33,401 REGIONS OF THE CEREBELLAR 751 00:32:33,401 --> 00:32:34,936 GRANULAR LAYER AT INDIVIDUAL 752 00:32:34,936 --> 00:32:39,640 PATIENT LEVEL, WHEN WE COMBINED 753 00:32:39,640 --> 00:32:40,841 ALL HIGH INFECTION SPOTS ACROSS 754 00:32:40,841 --> 00:32:43,277 ALL OF THE PATIENTS THAT WE 755 00:32:43,277 --> 00:32:45,313 EVALUATED, AND COMPARED THAT 756 00:32:45,313 --> 00:32:48,282 WITH MID-AND LOW INFECTION 757 00:32:48,282 --> 00:32:53,421 SPOTS, WE REVEAL A HANDFUL OF 758 00:32:53,421 --> 00:32:53,955 POTENTIAL -- PREDOMINANTLY 759 00:32:53,955 --> 00:32:56,591 NEURONAL SIGNALING GENES THAT 760 00:32:56,591 --> 00:32:58,693 WERE DIFFERENTIALLY INFECTED, 761 00:32:58,693 --> 00:33:00,761 BETWEEN HIGH AND MID-VERSUS LOW. 762 00:33:00,761 --> 00:33:02,730 NOW, THESE ARE PRELIMINARY DATA 763 00:33:02,730 --> 00:33:06,167 WHICH WE INTEND TO PURSUE 764 00:33:06,167 --> 00:33:07,501 FURTHER. 765 00:33:07,501 --> 00:33:09,904 WITH ADDITIONAL SEQUENCING 766 00:33:09,904 --> 00:33:12,506 APPROACHES AND ATTEMPT TO 767 00:33:12,506 --> 00:33:14,375 CROSS-VALIDATE PHENOTYPIC AND 768 00:33:14,375 --> 00:33:16,711 FUNCTIONAL IMPLICATIONS OF THESE 769 00:33:16,711 --> 00:33:17,178 DIFFERENT -- OF THIS 770 00:33:17,178 --> 00:33:22,149 DIFFERENTIAL GENE EXPRESSION? 771 00:33:22,149 --> 00:33:28,189 IN A HUMAN BRAINING OR ORGANOID 772 00:33:28,189 --> 00:33:30,324 MODEL, LET BY A POSTDOC, SOPHIA, 773 00:33:30,324 --> 00:33:31,459 IN THE AUDIENCE TODAY. 774 00:33:31,459 --> 00:33:35,162 IN CLOSING, I HOPE I CONVINCED 775 00:33:35,162 --> 00:33:37,431 YOU THAT AUTOPSY TOOLS ARE -- 776 00:33:37,431 --> 00:33:40,001 AUTOPSIES ARE A POWERFUL TOOL 777 00:33:40,001 --> 00:33:43,738 FOR GLEANING NEW INSIGHT INTO 778 00:33:43,738 --> 00:33:45,906 INFECTIOUS DISEASE PATHOGENESIS, 779 00:33:45,906 --> 00:33:49,310 AND THAT PROPER DESIGN AND 780 00:33:49,310 --> 00:33:52,246 SAMPLE HANDLING BEYOND WHAT IS 781 00:33:52,246 --> 00:33:55,082 ACCOMPLISHED IN STANDARD 782 00:33:55,082 --> 00:33:55,650 AUTOPSIES IS FUNDAMENTAL FOR 783 00:33:55,650 --> 00:33:58,386 SUCCESS, AND I HOPE I'VE 784 00:33:58,386 --> 00:34:01,022 CONVINCED YOU THAT THE NIH 785 00:34:01,022 --> 00:34:02,490 CLINICAL CENTER AND THE 786 00:34:02,490 --> 00:34:05,059 INTRAMURAL RESEARCH PROGRAM AT 787 00:34:05,059 --> 00:34:07,361 THE NIH ARE EXTREMELY WELL 788 00:34:07,361 --> 00:34:08,796 POSITIONED TO MEANINGFULLY 789 00:34:08,796 --> 00:34:11,198 ADVANCE THESE TYPES OF STUDIES 790 00:34:11,198 --> 00:34:14,535 FOR BOTH INFECTIOUS AND 791 00:34:14,535 --> 00:34:15,369 NON-INFECTIOUS DISEASES. 792 00:34:15,369 --> 00:34:17,872 WITH THAT I'M GOING TO TAKE TWO 793 00:34:17,872 --> 00:34:19,974 SECONDS TO JUST GO A LITTLE BIT 794 00:34:19,974 --> 00:34:21,042 OVER MY ACKNOWLEDGMENTS BECAUSE 795 00:34:21,042 --> 00:34:25,446 AS I SAID AT THE BEGINNING THIS 796 00:34:25,446 --> 00:34:26,747 REALLY WAS TEAM SCIENCE. 797 00:34:26,747 --> 00:34:28,949 AND WHILE I TRIED TO HIGHLIGHT A 798 00:34:28,949 --> 00:34:30,685 NUMBER OF INDIVIDUALS WHO 799 00:34:30,685 --> 00:34:31,886 CONTRIBUTED TO INDIVIDUAL 800 00:34:31,886 --> 00:34:33,421 COMPONENTS OF THIS, I'D LIKE TO 801 00:34:33,421 --> 00:34:34,955 JUST SPEND A MOMENT TO 802 00:34:34,955 --> 00:34:36,490 ACKNOWLEDGE A FEW PEOPLE. 803 00:34:36,490 --> 00:34:37,925 SO THESE STUDIES WOULD NOT HAVE 804 00:34:37,925 --> 00:34:39,160 BEEN POSSIBLE WITHOUT OUR 805 00:34:39,160 --> 00:34:39,894 RESEARCH VILLAGE. 806 00:34:39,894 --> 00:34:42,096 I'D LIKE TO THANK AND 807 00:34:42,096 --> 00:34:43,831 ACKNOWLEDGE LARGER NIH COVID 808 00:34:43,831 --> 00:34:46,467 AUTOPSY CONSORTIUM, CALL OUT THE 809 00:34:46,467 --> 00:34:48,135 NIH CLINICAL CENTER ADMISSIONS 810 00:34:48,135 --> 00:34:50,671 DEPARTMENT THAT TOOK LATE NIGHT 811 00:34:50,671 --> 00:34:52,306 PHONE CALLS AND FACILITATED 812 00:34:52,306 --> 00:34:56,444 TRANSFER OF OUR NEXT INDIVIDUALS 813 00:34:56,444 --> 00:34:57,111 FOR POSTMORTEM STUDIES. 814 00:34:57,111 --> 00:34:59,413 I'D LIKE TO ACKNOWLEDGE OUR 815 00:34:59,413 --> 00:35:01,248 CLOSE COLLABORATOR AND CO-LEADS 816 00:35:01,248 --> 00:35:04,085 IN THE NCI PATHOLOGY GROUP, DR. 817 00:35:04,085 --> 00:35:05,953 KLEINER AND HEWITT, AND ALL 818 00:35:05,953 --> 00:35:12,093 THEIR STAFF AND TRAINEES, 819 00:35:12,093 --> 00:35:13,527 ACKNOWLEDGE DR. SYDNEY STEIN 820 00:35:13,527 --> 00:35:16,030 FROM MY GROUP, AND FIRST AUTHOR 821 00:35:16,030 --> 00:35:17,898 ON "NATURE" STUDY AND OTHERS IN 822 00:35:17,898 --> 00:35:22,069 THE CRITICAL CARE MEDICINE 823 00:35:22,069 --> 00:35:24,271 DEPARTMENT PARTICULARLY SABRINA 824 00:35:24,271 --> 00:35:27,174 RAMELLI, FOR HER ENDLESS HOURS 825 00:35:27,174 --> 00:35:30,678 AND KEVIN VANILLA, WHO IS ALSO 826 00:35:30,678 --> 00:35:32,012 IN ATTENDANCE TODAY, FOR HELPING 827 00:35:32,012 --> 00:35:37,151 SO MUCH MOVE THIS WORK FORWARD, 828 00:35:37,151 --> 00:35:38,219 AND DR. ALLISON GRAZIOLI, LEAD 829 00:35:38,219 --> 00:35:39,320 CLINICAL PARTNER AT THE 830 00:35:39,320 --> 00:35:41,622 UNIVERSITY OF MARYLAND, AND SO 831 00:35:41,622 --> 00:35:43,157 MANY OF HER COLLEAGUES ACROSS 832 00:35:43,157 --> 00:35:48,195 UNIVERSITY OF MARYLAND SYSTEM 833 00:35:48,195 --> 00:35:51,699 WHO TIRELESSLY CARED FOR MANY OF 834 00:35:51,699 --> 00:35:53,434 THESE PATIENTS, ULTIMATELY 835 00:35:53,434 --> 00:35:55,970 FACILITATED RECRUITMENT OF 836 00:35:55,970 --> 00:35:57,271 PATIENTS FOR THIS STUDY. 837 00:35:57,271 --> 00:36:01,108 I'D LIKE TO ACKNOWLEDGE MY 838 00:36:01,108 --> 00:36:02,276 COLLEAGUES IN NIAID AND THEIR 839 00:36:02,276 --> 00:36:05,813 LABS FOR THE WORK THEY HELPED 840 00:36:05,813 --> 00:36:09,216 WITH THE VIROLOGY, KAREN 841 00:36:09,216 --> 00:36:10,718 PETERSON AND CLAYTON WINKLER 842 00:36:10,718 --> 00:36:12,052 EVALUATING WHAT'S HAPPENING IN 843 00:36:12,052 --> 00:36:14,188 THE BRAIN, ELY I MENTIONED 844 00:36:14,188 --> 00:36:15,890 HELPED WITH THE LONG READ 845 00:36:15,890 --> 00:36:24,665 SEQUENCING, I'D LIKE TO 846 00:36:24,665 --> 00:36:32,273 SPECIFICALLY CALL OUT JEON ORGU 847 00:36:32,273 --> 00:36:33,107 DIS 848 00:36:33,107 --> 00:36:39,480 THE FREDERICK TEAM, NIDCR, PETER 849 00:36:39,480 --> 00:36:43,851 AND BLAKE, AND CHI CHAN FROM 850 00:36:43,851 --> 00:36:44,218 NEI. 851 00:36:44,218 --> 00:36:45,252 WITH THAT I'LL CLOSE. 852 00:36:45,252 --> 00:36:47,021 THANK YOU FOR YOUR ATTENTION, 853 00:36:47,021 --> 00:36:49,223 AND I'LL TAKE ANY QUESTIONS YOU 854 00:36:49,223 --> 00:36:49,657 MAY HAVE. 855 00:36:49,657 --> 00:36:50,691 [APPLAUSE] 856 00:36:50,691 --> 00:36:52,293 >> THANK YOU. 857 00:36:52,293 --> 00:36:56,363 THAT WAS FABULOUS LECTURE. 858 00:36:56,363 --> 00:37:00,367 A REMINDER OF HOW AUTOPSIES AND 859 00:37:00,367 --> 00:37:02,436 PATHOLOGIES CAN REALLY ENHANCE 860 00:37:02,436 --> 00:37:04,305 AND EXTEND OUR KNOWLEDGE OF SOME 861 00:37:04,305 --> 00:37:05,639 VERY IMPORTANT DISEASES. 862 00:37:05,639 --> 00:37:06,841 I'D LIKE THE AUDIENCE TO GO 863 00:37:06,841 --> 00:37:09,910 AHEAD IF YOU HAVE QUESTIONS, GO 864 00:37:09,910 --> 00:37:11,879 AHEAD AND COME TO THE 865 00:37:11,879 --> 00:37:13,314 MICROPHONE, AND THOSE ONLINE I 866 00:37:13,314 --> 00:37:15,349 THINK IF YOU WOULD LIKE TO TYPE 867 00:37:15,349 --> 00:37:16,917 IN A QUESTION, IT WILL COME TO 868 00:37:16,917 --> 00:37:23,924 MY E-MAIL HERE AND I WILL ASK 869 00:37:23,924 --> 00:37:25,292 DAN THE THINGS YOU'RE INTERESTED 870 00:37:25,292 --> 00:37:25,459 IN. 871 00:37:25,459 --> 00:37:26,694 I'D LIKE TO TAKE THE OPPORTUNITY 872 00:37:26,694 --> 00:37:28,395 TO ASK THE FIRST QUESTION, DAN, 873 00:37:28,395 --> 00:37:32,132 WHICH IS I KNOW YOUR NUMBERS 874 00:37:32,132 --> 00:37:33,634 WERE SMALL, BUT WHAT CAN YOU 875 00:37:33,634 --> 00:37:37,271 TELL US ABOUT THE DEMOGRAPHICS 876 00:37:37,271 --> 00:37:40,107 OF YOUR COHORT, AND COULD YOU 877 00:37:40,107 --> 00:37:41,942 TELL US IF THERE'S ANY SORT OF 878 00:37:41,942 --> 00:37:45,813 IMPACT OF THOSE DEMOGRAPHICS ON 879 00:37:45,813 --> 00:37:48,782 YOUR FINDINGS, GENDER, AGE, 880 00:37:48,782 --> 00:37:49,316 CO-MORBIDITIES, YOU HAVE 881 00:37:49,316 --> 00:37:50,084 INTERESTING FINDINGS BUT WHAT 882 00:37:50,084 --> 00:37:51,952 ELSE CAN YOU TELL US ABOUT HOW 883 00:37:51,952 --> 00:37:53,821 THE INDIVIDUALS WHO ARE 884 00:37:53,821 --> 00:37:58,626 AUTOPSIES MIGHT HAVE AFFECTED 885 00:37:58,626 --> 00:38:00,327 YOUR RESULTS. 886 00:38:00,327 --> 00:38:01,462 >> GREAT QUESTION, DR. CHAN. 887 00:38:01,462 --> 00:38:05,399 ONE OF THE MOST IMPORTANT 888 00:38:05,399 --> 00:38:07,167 ASPECTS OF THE DEMOGRAPHICS AND 889 00:38:07,167 --> 00:38:11,539 COMORBIDITIES OF OUR COHORT 890 00:38:11,539 --> 00:38:16,110 RELATES, AND WE WERE CABLE ABOUT 891 00:38:16,110 --> 00:38:17,311 THIS, GENERALIZABILITY OF OUR 892 00:38:17,311 --> 00:38:17,645 FINDINGS. 893 00:38:17,645 --> 00:38:20,881 WE HAD AN OLDER AND SICKER 894 00:38:20,881 --> 00:38:24,351 COHORT, WILD TYPE VIRUS 895 00:38:24,351 --> 00:38:25,119 UNVACCINATED EARLY. 896 00:38:25,119 --> 00:38:27,121 WE SAW REALLY A SEVERE PHENOTYPE 897 00:38:27,121 --> 00:38:28,422 OF DISEASE, PARTICULARLY IN THAT 898 00:38:28,422 --> 00:38:29,857 PATIENT POPULATION. 899 00:38:29,857 --> 00:38:32,493 THERE'S NO DOUBT IN MY MIND, WE 900 00:38:32,493 --> 00:38:35,095 ALL KNOW THAT OVER TIME THIS 901 00:38:35,095 --> 00:38:36,730 DISEASE HAS CHANGED. 902 00:38:36,730 --> 00:38:38,532 SO WHAT IT EMPHASIZES WE NEED TO 903 00:38:38,532 --> 00:38:40,601 BE CAUTIOUS ABOUT HOW WE 904 00:38:40,601 --> 00:38:42,670 INTERPRET OUR FINDINGS FOR THIS 905 00:38:42,670 --> 00:38:43,070 COHORT. 906 00:38:43,070 --> 00:38:46,173 I WILL MAKE ONE COMMENT ABOUT, 907 00:38:46,173 --> 00:38:48,809 YOU KNOW, HOW MIGHT SOME OF OUR 908 00:38:48,809 --> 00:38:50,077 CLINICAL OR DEMOGRAPHIC 909 00:38:50,077 --> 00:38:50,911 VARIABLES PROVIDE SOME INSIGHT. 910 00:38:50,911 --> 00:38:52,846 ONE OF THE THINGS I DIDN'T TALK 911 00:38:52,846 --> 00:38:55,950 ABOUT IS THAT WE'RE ACTUALLY 912 00:38:55,950 --> 00:39:00,721 LOOKING AT DIFFERENCES IN THE 913 00:39:00,721 --> 00:39:02,156 T-CELL RECEPTOR BREADTH, DEPTH, 914 00:39:02,156 --> 00:39:02,990 SARS-COV-2 SPECIFICITY, NOT JUST 915 00:39:02,990 --> 00:39:08,963 IN THE LUNGS BUT ACTUALLY IN THE 916 00:39:08,963 --> 00:39:11,999 LYMPH NODE COMPARTMENT, SPLEEN 917 00:39:11,999 --> 00:39:12,666 COMPARTMENT AND PBMCs, I 918 00:39:12,666 --> 00:39:15,603 DIDN'T PRESENT THAT PART. 919 00:39:15,603 --> 00:39:16,837 ONE OF THE DEMOGRAPHIC VARIABLES 920 00:39:16,837 --> 00:39:19,974 THAT STUCK OUT IN THAT DATA WAS 921 00:39:19,974 --> 00:39:20,374 AGE. 922 00:39:20,374 --> 00:39:23,043 SO, THE T-CELL RESPONSES IN 923 00:39:23,043 --> 00:39:27,047 TISSUES ACROSS MULTIPLE OF OUR 924 00:39:27,047 --> 00:39:28,882 PARAMETERS, BOTH COV 2 SPECIFIC 925 00:39:28,882 --> 00:39:32,019 AND NON-VARIED AS A FUNCTION OF 926 00:39:32,019 --> 00:39:33,454 AGE, A TEASER FOR WHAT 927 00:39:33,454 --> 00:39:36,857 ULTIMATELY WILL COME OUT LATER. 928 00:39:36,857 --> 00:39:37,925 >> GREAT TALK. 929 00:39:37,925 --> 00:39:45,065 YOUR DATA SUGGESTS A WELL-TIMED 930 00:39:45,065 --> 00:39:46,367 IMMUNOSUPPRESSION MIGHT HELP. 931 00:39:46,367 --> 00:39:49,203 >> DR. OWENS, THANK YOU FOR THAT 932 00:39:49,203 --> 00:39:50,104 QUESTION. 933 00:39:50,104 --> 00:39:51,505 AND AS YOU KNOW, DURING THE 934 00:39:51,505 --> 00:39:54,475 EARLY STAGES OF THE PANDEMIC, 935 00:39:54,475 --> 00:39:57,311 THE NIH AND INTRAMURAL AND 936 00:39:57,311 --> 00:40:00,414 EXTRAMURAL, YOU KNOW, REALLY 937 00:40:00,414 --> 00:40:02,583 MADE SIGNIFICANT EFFORTS TO 938 00:40:02,583 --> 00:40:03,283 BRING MULTIPLE POTENTIAL 939 00:40:03,283 --> 00:40:04,318 THERAPEUTICS INTO CLINICAL 940 00:40:04,318 --> 00:40:06,754 TRIALS AND EVALUATE EFFICACY, 941 00:40:06,754 --> 00:40:09,223 AND A NUMBER OF THOSE HAVE SHOWN 942 00:40:09,223 --> 00:40:11,458 TO BE EFFICACIOUS IN THE SICKEST 943 00:40:11,458 --> 00:40:12,693 OF THE SICK INCLUDING STEROIDS 944 00:40:12,693 --> 00:40:14,962 AND A NUMBER OF OTHER HOST 945 00:40:14,962 --> 00:40:15,362 TARGET THERAPIES. 946 00:40:15,362 --> 00:40:17,164 NOW, WHAT WE KNOW ABOUT THOSE 947 00:40:17,164 --> 00:40:19,233 THERAPIES, AND LOGICALLY AND 948 00:40:19,233 --> 00:40:20,234 BIOLOGICALLY IT MAKES SENSE, IN 949 00:40:20,234 --> 00:40:23,404 THE CONTEXT OF OUR COHORT. 950 00:40:23,404 --> 00:40:25,606 SO, I'M GLAD YOU POINT IT OUT 951 00:40:25,606 --> 00:40:27,241 BECAUSE IF YOU'RE TARGETING ONLY 952 00:40:27,241 --> 00:40:31,078 THE VIRUS, BUT THE VIRUS STOPS 953 00:40:31,078 --> 00:40:32,346 REPLICATING, SOMEWHERE AROUND 954 00:40:32,346 --> 00:40:35,249 DAY 9 OR 14 BUT IN SEVEREST OF 955 00:40:35,249 --> 00:40:38,819 CASES, YOU KNOW, DISEASE 956 00:40:38,819 --> 00:40:39,386 PROGRESSES, MAYBE REPAIR IS 957 00:40:39,386 --> 00:40:40,487 DISORDERED, WE NEED TO BE 958 00:40:40,487 --> 00:40:42,022 THINKING ABOUT HOW WE CAN 959 00:40:42,022 --> 00:40:42,256 TARGET. 960 00:40:42,256 --> 00:40:45,392 SO, I THINK MANY OF OUR 961 00:40:45,392 --> 00:40:46,927 THERAPIES WERE EMPIRIC BUT THE 962 00:40:46,927 --> 00:40:48,696 CONCEPT BEHIND THIS WORK IS THAT 963 00:40:48,696 --> 00:40:51,198 MAYBE WE CAN GET INSIGHT INTO, 964 00:40:51,198 --> 00:40:53,434 YOU KNOW, THE SPECIFIC CELLULAR 965 00:40:53,434 --> 00:40:55,669 TARGET OR TARGETS, THE RIGHT 966 00:40:55,669 --> 00:40:57,304 TIMING, BIOMARKERS THAT ARE 967 00:40:57,304 --> 00:40:58,505 EASILY ACCESSIBLE IN THOSE 968 00:40:58,505 --> 00:40:59,173 INDIVIDUALS, THAT KIND OF GIVE 969 00:40:59,173 --> 00:41:00,808 YOU SENSE OF WHERE THEY ARE 970 00:41:00,808 --> 00:41:02,042 ALONG THE DISEASE COURSE, THAT'S 971 00:41:02,042 --> 00:41:07,981 ALL THE DIRECTION WE WANT TO GO. 972 00:41:07,981 --> 00:41:10,517 WE'RE JUST GETTING STARTED. 973 00:41:10,517 --> 00:41:11,719 >> THANK YOU. 974 00:41:11,719 --> 00:41:13,921 >> SO, DAN, WE HAVE A NUMBER OF 975 00:41:13,921 --> 00:41:16,757 ONLINE QUESTIONS. 976 00:41:16,757 --> 00:41:19,393 THE FIRST ONE IS, WHERE WERE THE 977 00:41:19,393 --> 00:41:22,362 AUTOPSIES PERFORMED AND DID THE 978 00:41:22,362 --> 00:41:23,030 CLINICIANS TAKE SPECIAL 979 00:41:23,030 --> 00:41:23,797 PRECAUTIONS? 980 00:41:23,797 --> 00:41:27,301 >> ALL OF THESE STUDIES WERE 981 00:41:27,301 --> 00:41:29,703 PERFORMED HERE WITHIN THE NCI 982 00:41:29,703 --> 00:41:31,138 PATHOLOGY SUITE. 983 00:41:31,138 --> 00:41:33,974 OUR PRIMARY LEADS ON THIS WERE 984 00:41:33,974 --> 00:41:35,609 STEPHEN HEWITT, HERE WITH US 985 00:41:35,609 --> 00:41:38,912 TODAY, CO-LEAD ON OUR STUDY, AND 986 00:41:38,912 --> 00:41:39,546 DAVID KLEINER. 987 00:41:39,546 --> 00:41:41,415 SO THEY WERE ALL DONE HERE. 988 00:41:41,415 --> 00:41:45,119 AT THAT STAGE OF THE PANDEMIC, 989 00:41:45,119 --> 00:41:46,987 AGAIN, NO VACCINES, SEVERE 990 00:41:46,987 --> 00:41:48,522 PHENOTYPE, WE ALL WORE 991 00:41:48,522 --> 00:41:52,893 ESSENTIALLY WHAT WOULD BE 992 00:41:52,893 --> 00:41:54,995 CLASSIFIED AS BIOSAFETY LEVEL 2 993 00:41:54,995 --> 00:41:56,730 WITH BIOSAFETY LEVEL 3 994 00:41:56,730 --> 00:42:00,901 PRECAUTIONS, SO WE HAD TYVEK 995 00:42:00,901 --> 00:42:03,604 SUITS, GOWNS, GLOVES, TO PREVENT 996 00:42:03,604 --> 00:42:03,871 EXPOSURES. 997 00:42:03,871 --> 00:42:06,206 I THINK VERONICA MAY HAVE HAD -- 998 00:42:06,206 --> 00:42:09,643 >> YEAH, I APPRECIATE, I'M 999 00:42:09,643 --> 00:42:12,246 NOT -- THIS IS OUTSIDE MY FIELD 1000 00:42:12,246 --> 00:42:13,714 BUT SO FASCINATING, THOUGHT YOU 1001 00:42:13,714 --> 00:42:18,852 DID A REALLY GOOD JOB EXPLAINING 1002 00:42:18,852 --> 00:42:20,053 THIS STUDY. 1003 00:42:20,053 --> 00:42:23,323 BEAR WITH ME BECAUSE I CLEAR 1004 00:42:23,323 --> 00:42:24,458 LIAM NOT VERY KNOWLEDGEABLE. 1005 00:42:24,458 --> 00:42:27,394 MY QUESTION IS A LITTLE BIT OF A 1006 00:42:27,394 --> 00:42:36,003 FOLLOW-UP OF DR. OWENS TRYING TO 1007 00:42:36,003 --> 00:42:37,104 UNDERSTAND PROLONGED IMMUNE 1008 00:42:37,104 --> 00:42:37,371 RESPONSE. 1009 00:42:37,371 --> 00:42:39,072 BECAUSE YOU'RE DEALING WITH 1010 00:42:39,072 --> 00:42:40,707 AUTOPSIES, IT'S EASIER TO GET 1011 00:42:40,707 --> 00:42:42,810 ACCESS TO DNA SU INTO THERE ANY 1012 00:42:42,810 --> 00:42:46,613 WAY OF GETTING ACCESS TO 1013 00:42:46,613 --> 00:42:47,181 PROTEINS? 1014 00:42:47,181 --> 00:42:48,649 I WONDER DOES PROTEINS FROM THE 1015 00:42:48,649 --> 00:42:51,518 VIRUS, THEY ARE STILL PRESENT IN 1016 00:42:51,518 --> 00:42:52,452 SOME PATIENTS PROLONGED IN THE 1017 00:42:52,452 --> 00:42:54,855 IMMUNE RESPONSE, WHAT DO YOU 1018 00:42:54,855 --> 00:42:59,326 THINK IS THE MECHANISM YOU 1019 00:42:59,326 --> 00:43:00,427 DEMONSTRATED IN THE PATIENTS 1020 00:43:00,427 --> 00:43:02,963 THAT END UP WITH DEATH THERE IS 1021 00:43:02,963 --> 00:43:05,032 THIS PROLONGATION OF IMMUNE 1022 00:43:05,032 --> 00:43:06,233 RESPONSE BUT ONTO THE -- HOW 1023 00:43:06,233 --> 00:43:08,735 THIS IS CARRIED BY THE VIRUS 1024 00:43:08,735 --> 00:43:09,837 SEEMS TO BE TRANSFERRED THROUGH 1025 00:43:09,837 --> 00:43:14,541 THE BLOOD AND ALL THAT, BUT WITH 1026 00:43:14,541 --> 00:43:15,776 RELATES TO LATE ONGOING IMMUNE 1027 00:43:15,776 --> 00:43:17,177 RESPONSE YOU DO YOU HAVE ANY 1028 00:43:17,177 --> 00:43:21,114 MECHANISMS THAT YOU CAN PROPOSE? 1029 00:43:21,114 --> 00:43:23,417 >> I HAVE THOUGHTS ABOUT WHAT 1030 00:43:23,417 --> 00:43:24,718 MIGHT BE PROPOSED. 1031 00:43:24,718 --> 00:43:27,287 AND THAT'S THE MOST IMPORTANT 1032 00:43:27,287 --> 00:43:29,423 THING BECAUSE WE HAVE NOT -- 1033 00:43:29,423 --> 00:43:31,291 WE'VE MADE OBSERVATION, WE'VE 1034 00:43:31,291 --> 00:43:34,695 NOT SHOWN OR PROVEN MECHANISMS, 1035 00:43:34,695 --> 00:43:37,865 SO WE ARE VERY CAUTIOUS. 1036 00:43:37,865 --> 00:43:40,400 YOUR QUESTION GETS TO THE HEART, 1037 00:43:40,400 --> 00:43:41,802 WE'RE ALSO CAREFUL TO SAY THAT 1038 00:43:41,802 --> 00:43:44,204 IN THIS STUDY WE DIDN'T STUDY 1039 00:43:44,204 --> 00:43:46,607 LONG COVID, RIGHT? 1040 00:43:46,607 --> 00:43:49,009 WE STUDIED INDIVIDUALS THAT HAD 1041 00:43:49,009 --> 00:43:51,311 SEVERE, YOU KNOW, MAJORITY OF 1042 00:43:51,311 --> 00:43:53,180 THEM HAD SEVERE ACUTE COVID, 1043 00:43:53,180 --> 00:43:55,148 MAJORITY OF OUR COHORT DIED FROM 1044 00:43:55,148 --> 00:43:57,084 SEVERE ACUTE COVID. 1045 00:43:57,084 --> 00:43:59,953 THERE ARE A FEW THAT RECOVERED 1046 00:43:59,953 --> 00:44:03,457 AND THEN WENT ON TO DIE, YOU 1047 00:44:03,457 --> 00:44:07,594 KNOW, AFTER RECOVERING FROM 1048 00:44:07,594 --> 00:44:09,363 COVID, BUT IMPORTANTLY THOSE 1049 00:44:09,363 --> 00:44:11,531 INDIVIDUALS DIDN'T MEET ANY 1050 00:44:11,531 --> 00:44:12,866 CLINICAL SYNDROMIC CRITERIA FOR 1051 00:44:12,866 --> 00:44:14,067 QUOTE/UNQUOTE LONG COVID. 1052 00:44:14,067 --> 00:44:16,703 NOW, WITH THAT IN MIND, YOU 1053 00:44:16,703 --> 00:44:19,740 KNOW, SO WE'RE CAREFUL NOT TO 1054 00:44:19,740 --> 00:44:22,476 SAY THAT OUR STUDY DIRECTLY 1055 00:44:22,476 --> 00:44:23,677 INFORMS LONG COVID. 1056 00:44:23,677 --> 00:44:28,282 WHAT WE DO SAY IS THAT OUR STUDY 1057 00:44:28,282 --> 00:44:29,750 PROVIDES PROOF OF PRINCIPLE THAT 1058 00:44:29,750 --> 00:44:32,586 PERHAPS IN ANOTHER, YOU KNOW, IN 1059 00:44:32,586 --> 00:44:36,723 THIS OTHER DISEASE ENTITY THAT 1060 00:44:36,723 --> 00:44:39,126 PERSISTENCE OF VIRAL COMPONENTS 1061 00:44:39,126 --> 00:44:42,930 MIGHT BE TRIGGERING OR MIGHT BE 1062 00:44:42,930 --> 00:44:44,264 CONTRIBUTING TO PROLONGED 1063 00:44:44,264 --> 00:44:45,232 INFLAMMATORY RESPONSE. 1064 00:44:45,232 --> 00:44:48,001 THEY MIGHT, YOU KNOW, OTHER 1065 00:44:48,001 --> 00:44:53,006 ASPECTS OF THE LONG COVID STORY, 1066 00:44:53,006 --> 00:44:56,410 ARE THEY RESULTING IN 1067 00:44:56,410 --> 00:44:57,611 RECREDESENCE OF OTHER VIRUSES, 1068 00:44:57,611 --> 00:44:59,112 HERPES VIRUSES, THAT MIGHT BE 1069 00:44:59,112 --> 00:45:01,114 CONTRIBUTING TO SOME OF THE 1070 00:45:01,114 --> 00:45:06,053 CLINICAL MANIFESTATIONS THAT ARE 1071 00:45:06,053 --> 00:45:08,121 OBSERVED IN LONG COVID. 1072 00:45:08,121 --> 00:45:10,857 I DON'T KNOW THAT YOU ABOUT WE 1073 00:45:10,857 --> 00:45:11,725 PROVIDE BIOLOGICAL 1074 00:45:11,725 --> 00:45:15,062 POSSIBILITIES, AND OTHERS, WE'RE 1075 00:45:15,062 --> 00:45:19,967 NOT THE ONLY ONES, IN HUMAN 1076 00:45:19,967 --> 00:45:21,101 STUDIES WHERE WE COLLECTED 1077 00:45:21,101 --> 00:45:25,872 TISSUES, IN THE G.I. TRACT, WERE 1078 00:45:25,872 --> 00:45:28,175 RNA OR PROTEINS IN INDIVIDUALS 1079 00:45:28,175 --> 00:45:31,678 THAT MEET SYNDROMIC CRITERIA WE 1080 00:45:31,678 --> 00:45:32,446 PROVIDED ADEQUATE BIOLOGIC 1081 00:45:32,446 --> 00:45:33,647 RATIONALE WE NEED TO CONSIDER 1082 00:45:33,647 --> 00:45:37,117 THAT, PERSISTENCE OF VIRAL 1083 00:45:37,117 --> 00:45:39,086 REMNANTS AND OTHER PROLONGED 1084 00:45:39,086 --> 00:45:40,187 AND/OR DISORDERED IMMUNE 1085 00:45:40,187 --> 00:45:41,221 RESPONSE CONTRIBUTING TO 1086 00:45:41,221 --> 00:45:41,755 CLINICAL MANIFESTATIONS. 1087 00:45:41,755 --> 00:45:43,023 WE DON'T HAVE ANSWERS BUT I 1088 00:45:43,023 --> 00:45:47,427 THINK WE HAVE RAISED SOME 1089 00:45:47,427 --> 00:45:48,395 ADDITIONAL QUESTIONS. 1090 00:45:48,395 --> 00:45:49,529 >> WE HAVE A NUMBER OF OTHER 1091 00:45:49,529 --> 00:45:50,364 ONLINE QUESTIONS. 1092 00:45:50,364 --> 00:45:53,567 WERE YOU ABLE TO VALIDATE SG RNA 1093 00:45:53,567 --> 00:45:56,503 FOUND IN THE GREATER THAN 31-DAY 1094 00:45:56,503 --> 00:45:59,272 COHORT IN THE SMALL INTESTINE 1095 00:45:59,272 --> 00:46:00,807 TISSUES? 1096 00:46:00,807 --> 00:46:03,977 >> SO, IF I WENT BACK TO MY HEAT 1097 00:46:03,977 --> 00:46:06,680 MAP, WHICH IS IN THERE, WHAT 1098 00:46:06,680 --> 00:46:11,952 I'LL SAY IS THAT THE MAJORITY OF 1099 00:46:11,952 --> 00:46:14,254 OUR SUBGENOMIC RNA POSITIVE 1100 00:46:14,254 --> 00:46:17,958 SAMPLES ACROSS RESPIRATORY AND 1101 00:46:17,958 --> 00:46:18,525 NON-RESPIRATORY TISSUES 1102 00:46:18,525 --> 00:46:20,260 PREDOMINATED IN OUR 1103 00:46:20,260 --> 00:46:21,561 QUOTE/UNQUOTE EARLY COHORT. 1104 00:46:21,561 --> 00:46:23,964 WE DEFINED OUR EARLY COHORT AS 1105 00:46:23,964 --> 00:46:27,267 LESS THAN OR EQUAL TO 14 DAYS 1106 00:46:27,267 --> 00:46:28,035 AFTER, YOU KNOW, 1107 00:46:28,035 --> 00:46:29,770 PATIENT-REPORTED ILLNESS ONSET. 1108 00:46:29,770 --> 00:46:31,738 SO THE MAJORITY OF RESPIRATORY 1109 00:46:31,738 --> 00:46:33,607 AND NON-RESPIRATORY SAMPLES 1110 00:46:33,607 --> 00:46:35,142 POSITIVE FOR SUBGENOMIC RNA FELL 1111 00:46:35,142 --> 00:46:37,577 IN EARLY COHORT. 1112 00:46:37,577 --> 00:46:40,747 NOW, THERE ARE A HANDFUL OF 1113 00:46:40,747 --> 00:46:44,117 INDIVIDUALS AND A HANDFUL OF 1114 00:46:44,117 --> 00:46:48,388 TISSUES MORE SPORADICALLY WHERE 1115 00:46:48,388 --> 00:46:49,923 WE FOUND EVIDENCE OF SUBGENOMIC 1116 00:46:49,923 --> 00:46:51,691 RNA LATER IN THE TIME COURSE. 1117 00:46:51,691 --> 00:46:53,627 ONE THAT WAS PARTICULARLY 1118 00:46:53,627 --> 00:46:55,695 INTERESTING, NOW WELL KNOWN AND 1119 00:46:55,695 --> 00:46:58,231 MORE OR LESS ACCEPTED IN THE 1120 00:46:58,231 --> 00:47:03,336 FIELD, IS WHAT IS PATIENT 33 IN 1121 00:47:03,336 --> 00:47:03,837 OUR COHORT. 1122 00:47:03,837 --> 00:47:06,206 PATIENT 33 DIED 76 DAYS AFTER 1123 00:47:06,206 --> 00:47:07,541 ILLNESS ONSET, THEIR CLINICAL 1124 00:47:07,541 --> 00:47:09,943 HISTORY WAS INTERESTING. 1125 00:47:09,943 --> 00:47:13,213 THEY HAD PREEXISTING LUNG 1126 00:47:13,213 --> 00:47:14,781 DISEASE, THEY HAD CO-MORBIDITIES 1127 00:47:14,781 --> 00:47:16,750 THEY HAD ACUTE COVID, AT THAT 1128 00:47:16,750 --> 00:47:19,052 TIME PATIENTS WERE BEING TREATED 1129 00:47:19,052 --> 00:47:20,587 WITH STEROIDS FOR SEVERE ACUTE 1130 00:47:20,587 --> 00:47:21,688 COVID. 1131 00:47:21,688 --> 00:47:23,423 IT WAS A CLINICAL OBSERVATION 1132 00:47:23,423 --> 00:47:24,758 THAT OFTEN WHEN YOU WITHDREW 1133 00:47:24,758 --> 00:47:26,960 THOSE STEROIDS WHEN YOU TAPERED 1134 00:47:26,960 --> 00:47:29,129 THEM, THE DISEASE WOULD FLARE. 1135 00:47:29,129 --> 00:47:29,963 SO THIS PARTICULAR INDIVIDUAL 1136 00:47:29,963 --> 00:47:32,099 WAS NOT ABLE TO BE TAPERED OFF 1137 00:47:32,099 --> 00:47:33,300 THOSE STEROIDS. 1138 00:47:33,300 --> 00:47:38,038 AND WAS ON A PROLONGED COURSE OF 1139 00:47:38,038 --> 00:47:40,874 STEROIDS AND DIED 76 DAYS AFTER, 1140 00:47:40,874 --> 00:47:50,083 EVIDENCE OF SUBGENOMIC RNA AND 1141 00:47:50,083 --> 00:47:50,851 NON-RESPIRATORY COMPONENTS, 1142 00:47:50,851 --> 00:47:54,488 THERE'S POTENTIAL FOR ONGOING 1143 00:47:54,488 --> 00:47:57,557 VIRAL REPRESENTLY TAIGS IN 1144 00:47:57,557 --> 00:48:05,432 REMINDERS AND NON-RESPIRATORY 1145 00:48:05,432 --> 00:48:06,199 COMPARTMENTS. 1146 00:48:06,199 --> 00:48:08,935 >> WERE THERE CLINICAL FEATURES, 1147 00:48:08,935 --> 00:48:11,138 SEVERITY OF ILLNESS, ET CETERA. 1148 00:48:11,138 --> 00:48:13,240 >> YES, IF I WERE -- YOU'RE 1149 00:48:13,240 --> 00:48:15,876 STRETCHING MY MEMORY A LITTLE 1150 00:48:15,876 --> 00:48:16,042 BIT. 1151 00:48:16,042 --> 00:48:18,778 THE ONE STANDS OUT, OBVIOUSLY, 1152 00:48:18,778 --> 00:48:20,580 THAT THE PATIENT 27 I POINTED 1153 00:48:20,580 --> 00:48:25,051 OUT IN MY TALK, THAT INDIVIDUAL 1154 00:48:25,051 --> 00:48:25,685 PRESENTED QUOTE/UNQUOTE ONE DAY 1155 00:48:25,685 --> 00:48:27,988 AFTER ILLNESS ONSET. 1156 00:48:27,988 --> 00:48:30,757 NOW, REMEMBER, ILLNESS ONSET FOR 1157 00:48:30,757 --> 00:48:32,926 US IS SOMEWHAT OF A CLINICAL 1158 00:48:32,926 --> 00:48:36,129 ANCHOR BUT IT IS NOT LIKE AN 1159 00:48:36,129 --> 00:48:36,997 EXPERIMENTAL CONDITION. 1160 00:48:36,997 --> 00:48:38,765 YOU KNOW, IT'S SUBJECT TO BIAS, 1161 00:48:38,765 --> 00:48:40,934 SUBJECT TO PATIENT REPORT. 1162 00:48:40,934 --> 00:48:43,136 AND IT'S ALSO -- IT DOESN'T 1163 00:48:43,136 --> 00:48:46,206 REFLECT TIMING OF INFECTION. 1164 00:48:46,206 --> 00:48:47,807 OF COURSE THERE'S INCUBATION 1165 00:48:47,807 --> 00:48:50,477 PERIOD WITH SARS-COV-2, SO THAT 1166 00:48:50,477 --> 00:48:51,778 INDIVIDUAL IS PROBABLY INFECTED, 1167 00:48:51,778 --> 00:48:54,414 YOU KNOW, AGAIN GUESSING FIVE OR 1168 00:48:54,414 --> 00:48:57,050 SEVEN OR EIGHT DAYS BEFORE THEY 1169 00:48:57,050 --> 00:48:58,485 HAD SYMPTOMS. 1170 00:48:58,485 --> 00:49:00,887 BUT DIRECTLY TO ANSWER THE 1171 00:49:00,887 --> 00:49:04,257 QUESTION, TIMING OF PRESENTATION 1172 00:49:04,257 --> 00:49:06,459 AFTER ILLNESS ONSET WAS A 1173 00:49:06,459 --> 00:49:10,063 DEFINING FEATURE OF INDIVIDUALS 1174 00:49:10,063 --> 00:49:11,498 THAT WERE COMPLETELY 1175 00:49:11,498 --> 00:49:12,098 SERONEGATIVE. 1176 00:49:12,098 --> 00:49:13,366 >> IF I RECALL CORRECTLY THERE 1177 00:49:13,366 --> 00:49:15,902 WAS ONE PATIENT THAT HAD A VERY 1178 00:49:15,902 --> 00:49:17,737 HIGH VIRAL LOAD IN PLASMA BUT 1179 00:49:17,737 --> 00:49:20,140 SEEMED TO HAVE NO ANTIBODY 1180 00:49:20,140 --> 00:49:20,407 RESPONSE. 1181 00:49:20,407 --> 00:49:22,242 WAS THIS A KINETIC ISSUE OR 1182 00:49:22,242 --> 00:49:23,877 DEMONSTRATION OF THE ROLE THAT 1183 00:49:23,877 --> 00:49:25,445 ANTIBODIES PLAY IN CONTROL? 1184 00:49:25,445 --> 00:49:27,180 HOW WAS THIS FOLLOWED UP? 1185 00:49:27,180 --> 00:49:29,983 >> YEAH, VERY GOOD QUESTION. 1186 00:49:29,983 --> 00:49:34,588 THAT'S PATIENT 27. 1187 00:49:34,588 --> 00:49:36,323 WE HYPOTHESIZE THAT INDIVIDUAL 1188 00:49:36,323 --> 00:49:38,325 PRESENTED TOO EARLY AFTER 1189 00:49:38,325 --> 00:49:40,060 ILLNESS ONSET, TOO EARLY AFTER 1190 00:49:40,060 --> 00:49:50,237 INFECTION FOR THEM TO HAVE AN 1191 00:49:50,237 --> 00:49:52,772 ADEQUATE HUMOROAL RESPONSE. 1192 00:49:52,772 --> 00:49:58,445 THE ANTIBODY MEDIATED ARM, 1193 00:49:58,445 --> 00:50:04,584 ANTIBODIES ARE QUITE GOOD, 1194 00:50:04,584 --> 00:50:06,019 OBSINISING VIRUS. 1195 00:50:06,019 --> 00:50:06,886 WE HYPOTHESIZE IN THOSE 1196 00:50:06,886 --> 00:50:11,725 INDIVIDUALS THAT PRESENTED EARLY 1197 00:50:11,725 --> 00:50:15,428 BEFORE THE ONSET OF HUMORAL 1198 00:50:15,428 --> 00:50:17,931 RESPONSE, VIRUS CAN CIRCULATE. 1199 00:50:17,931 --> 00:50:21,101 ONCE YOU HAVE AD QUIT HUMORAL 1200 00:50:21,101 --> 00:50:23,637 RESPONSE, ADEQUATE TITER AND 1201 00:50:23,637 --> 00:50:26,473 AFFINITY, CIRCULATING VIRUS IS 1202 00:50:26,473 --> 00:50:28,108 PRESUMABLY OBSINISING, CANNOT GO 1203 00:50:28,108 --> 00:50:33,246 ON TO AFFECT CELLS AND TISSUES 1204 00:50:33,246 --> 00:50:35,448 OUTSIDE RESPIRATORY TRACT. 1205 00:50:35,448 --> 00:50:37,517 >> THANKS FOR BRILLIANT WORK, 1206 00:50:37,517 --> 00:50:39,052 REALLY OUTSTANDING LECTURE. 1207 00:50:39,052 --> 00:50:41,254 I'M CURIOUS ON SOMETHING SIMPLE, 1208 00:50:41,254 --> 00:50:43,590 IN THIS COHORT WHAT WAS THE 1209 00:50:43,590 --> 00:50:45,592 DISTRIBUTION OF DISEASE IN THE 1210 00:50:45,592 --> 00:50:50,096 LUNGS, WAS IT FOCAL OR WAS IT 1211 00:50:50,096 --> 00:50:52,599 DIFFUSE, BOTH? 1212 00:50:52,599 --> 00:50:58,305 >> YEAH, THE ANSWER IS THAT IT 1213 00:50:58,305 --> 00:50:59,072 WAS DIFFUSE. 1214 00:50:59,072 --> 00:51:01,474 YEAH, THIS IS -- IT'S AN 1215 00:51:01,474 --> 00:51:03,143 OPPORTUNITY TO HIGHLIGHT ANOTHER 1216 00:51:03,143 --> 00:51:05,111 POINT WHICH IS AN INTERESTING 1217 00:51:05,111 --> 00:51:08,982 POINT IS THAT IN THE LUNG WHERE 1218 00:51:08,982 --> 00:51:11,718 WE UNDERSTAND AND BELIEVE THAT 1219 00:51:11,718 --> 00:51:13,219 VIRUS SPREADS CONTIGUOUSLY, YOU 1220 00:51:13,219 --> 00:51:14,421 KNOW, YOU'LL SEE WHOLE REGIONS 1221 00:51:14,421 --> 00:51:16,389 OF AIRWAY AND WHOLE REGIONS OF 1222 00:51:16,389 --> 00:51:19,693 LUNG PARENCHYMA THAT ARE 1223 00:51:19,693 --> 00:51:19,993 INFECTED. 1224 00:51:19,993 --> 00:51:22,195 IN OTHER ORGANS WHERE THE 1225 00:51:22,195 --> 00:51:32,372 HYPOTHESIS IS THAT THERE'S, YOU 1226 00:51:32,372 --> 00:51:34,808 KNOW, HENOTOGENOU, SEEDING 1227 00:51:34,808 --> 00:51:36,476 THAT'S NOT OBSERVED OUTSIDE THE 1228 00:51:36,476 --> 00:51:36,976 LUNG. 1229 00:51:36,976 --> 00:51:40,780 THE BRAIN, FOR EXAMPLE, WE HAD 1230 00:51:40,780 --> 00:51:44,184 INFECTION OF MULTIPLE REGIONS OF 1231 00:51:44,184 --> 00:51:45,151 THE BRAIN NOT ANATOMICALLY 1232 00:51:45,151 --> 00:51:46,386 CONTIGUOUS, EVEN IN THOSE 1233 00:51:46,386 --> 00:51:51,391 INDIVIDUAL REGIONS IT WASN'T 1234 00:51:51,391 --> 00:51:58,531 LIKE THE WHOLE SECTION WAS 1235 00:51:58,531 --> 00:52:00,500 POSITIVE FOR ISH. 1236 00:52:00,500 --> 00:52:01,901 NEIGHBORING CLUSTERS WOULD BE 1237 00:52:01,901 --> 00:52:02,268 INFECTED. 1238 00:52:02,268 --> 00:52:04,437 I USE THE ANALOGY OF OUR PLAQUE 1239 00:52:04,437 --> 00:52:04,838 ASSAY. 1240 00:52:04,838 --> 00:52:11,111 WHEN WE EXPOSE A CELL LAYER TO 1241 00:52:11,111 --> 00:52:14,280 VIRUS, A NUMBER OF CELLS BECOME 1242 00:52:14,280 --> 00:52:14,948 INFECTED, VIRUS SPREADS 1243 00:52:14,948 --> 00:52:16,249 CONTIGUOUSLY, YOU GET PLAQUES OF 1244 00:52:16,249 --> 00:52:17,984 VIRUS BUT NOT THE WHOLE 1245 00:52:17,984 --> 00:52:19,018 MONOLAYER IS INFECTED. 1246 00:52:19,018 --> 00:52:21,421 ONLY LATER IF THE VIRUS, YOU 1247 00:52:21,421 --> 00:52:24,157 KNOW, REPLICATES IN THOSE CELLS, 1248 00:52:24,157 --> 00:52:25,458 THERE'S ABSENCE OF INTERFERON 1249 00:52:25,458 --> 00:52:27,961 RESPONSE YOU GET MORE CONFLUENT 1250 00:52:27,961 --> 00:52:28,962 CONTIGUOUS SPREAD. 1251 00:52:28,962 --> 00:52:29,729 IN MANY TISSUES INCLUDING THE 1252 00:52:29,729 --> 00:52:31,598 BRAIN THAT'S NOT WHAT WE 1253 00:52:31,598 --> 00:52:32,132 OBSERVED. 1254 00:52:32,132 --> 00:52:34,667 WE OBSERVED MORE OF THESE WHAT I 1255 00:52:34,667 --> 00:52:36,169 WOULD CALL PLAQUES RATHER THAN, 1256 00:52:36,169 --> 00:52:36,936 YOU KNOW, CONTIGUOUS SPREAD BUT 1257 00:52:36,936 --> 00:52:40,673 IN THE LUNG, YEAH, THE LUNG DOES 1258 00:52:40,673 --> 00:52:41,040 GREAT. 1259 00:52:41,040 --> 00:52:43,643 >> SO, THERE'S A NUMBER OF 1260 00:52:43,643 --> 00:52:46,479 QUESTIONS HERE ABOUT NEURONAL 1261 00:52:46,479 --> 00:52:48,882 INVOLVEMENT. 1262 00:52:48,882 --> 00:52:50,283 AND QUESTIONS RELATED TO WHETHER 1263 00:52:50,283 --> 00:52:54,988 OR NOT NEURONS EXPRESS AC 2, AND 1264 00:52:54,988 --> 00:52:57,424 IF NOT WAS THE VIRUS MEDIATED 1265 00:52:57,424 --> 00:53:02,996 THROUGH AC 2, IF NOT HOW WOULD 1266 00:53:02,996 --> 00:53:04,898 IT BE SPREAD THROUGHOUT THE 1267 00:53:04,898 --> 00:53:08,001 NEURONAL AREAS? 1268 00:53:08,001 --> 00:53:09,436 >> SO, ACE-2 IS EXPRESSED ON 1269 00:53:09,436 --> 00:53:12,071 NEURONS, THAT WAS KNOWN BEFORE, 1270 00:53:12,071 --> 00:53:14,908 IT WAS KNOWN AFTER COVID. 1271 00:53:14,908 --> 00:53:19,846 THE EXACT MECHANISM BY WHICH 1272 00:53:19,846 --> 00:53:20,847 VIRUS INFECTS NEURONS, I'M -- 1273 00:53:20,847 --> 00:53:23,883 THAT WORK MAY HAVE BEEN DONE, WE 1274 00:53:23,883 --> 00:53:25,418 DIDN'T DO THAT WORK BUT, AGAIN, 1275 00:53:25,418 --> 00:53:28,288 THAT WORK WOULD HAVE HAD TO HAVE 1276 00:53:28,288 --> 00:53:33,293 BEEN MODELED IN EITHER 2D OR 3D 1277 00:53:33,293 --> 00:53:34,160 NEURONAL SYSTEMS LIKE ORGANOIDS 1278 00:53:34,160 --> 00:53:36,029 OR OTHER SYSTEMS WHERE THEY CAN 1279 00:53:36,029 --> 00:53:38,398 DO, YOU KNOW, TARGETED KNOCKOUTS 1280 00:53:38,398 --> 00:53:39,866 OF DIFFERENT RECEPTORS, THOSE 1281 00:53:39,866 --> 00:53:42,602 TYPES OF THINGS. 1282 00:53:42,602 --> 00:53:45,104 SO, YES, NEURONS HAVE ACE-2. 1283 00:53:45,104 --> 00:53:49,275 WHAT THE MECHANISM OF INFECTION 1284 00:53:49,275 --> 00:53:51,277 OF NEURONS IS, I'M NOT -- I 1285 00:53:51,277 --> 00:53:52,579 DON'T THINK I KNOW THAT. 1286 00:53:52,579 --> 00:53:54,147 I KNOW I DON'T KNOW THAT ANSWER. 1287 00:53:54,147 --> 00:53:56,416 BUT THE OTHER PART OF THAT 1288 00:53:56,416 --> 00:53:57,617 QUESTION THAT'S IMPORTANT IS 1289 00:53:57,617 --> 00:54:00,153 EVEN IF A VIRUS INFECTS A CELL, 1290 00:54:00,153 --> 00:54:02,455 NOT ALL CELLS ARE CREATED EQUAL 1291 00:54:02,455 --> 00:54:05,391 SO SOME CELLS WILL UNDERGO 1292 00:54:05,391 --> 00:54:07,794 CYTOPATHIC EFFECT, HAVE A 1293 00:54:07,794 --> 00:54:08,461 RESPONSE. 1294 00:54:08,461 --> 00:54:10,096 SOME CELLS WILL GENERATE 1295 00:54:10,096 --> 00:54:11,965 INTERFERON RESPONSE THAT WILL 1296 00:54:11,965 --> 00:54:13,366 PRECLUDE VIRAL BUDDING AND 1297 00:54:13,366 --> 00:54:14,467 SPREAD TO NEIGHBORING CELLS, AND 1298 00:54:14,467 --> 00:54:16,636 SO JUST BECAUSE A CELL GETS 1299 00:54:16,636 --> 00:54:17,604 INFECTED THEY ARE NOT ALL GOING 1300 00:54:17,604 --> 00:54:18,638 TO ACT THE SAME WAY. 1301 00:54:18,638 --> 00:54:23,510 NEURONS SEEMED TO BE INFECTED AT 1302 00:54:23,510 --> 00:54:25,512 A RELATIVELY LOW LEVEL AND DON'T 1303 00:54:25,512 --> 00:54:28,448 AT LEAST BASED UPON OUR DATA 1304 00:54:28,448 --> 00:54:32,185 APPEAR TO UNDERGO CELL DEATH. 1305 00:54:32,185 --> 00:54:33,586 >> YOU MENTIONED INTERFERON. 1306 00:54:33,586 --> 00:54:35,588 I WAS WONDERING IF YOU KNOW THE 1307 00:54:35,588 --> 00:54:39,425 STATUS OF YOUR PATIENTS AND DID 1308 00:54:39,425 --> 00:54:40,326 THEY HAVE ANTI-TYPE I INTERFERON 1309 00:54:40,326 --> 00:54:43,162 ABC BECAUSE THAT WOULD MAKE THEM 1310 00:54:43,162 --> 00:54:44,163 EXTREME ARE -- ANTIBODIES 1311 00:54:44,163 --> 00:54:49,168 BECAUSE THAT WOULD MAKE THEM 1312 00:54:49,168 --> 00:54:49,636 EXTREMELY SUSCEPTIBLE. 1313 00:54:49,636 --> 00:54:52,705 >> WE DIDN'T PUBLISH BUT WE 1314 00:54:52,705 --> 00:54:55,441 WORKED WITH STEVE HOLLINS' 1315 00:54:55,441 --> 00:54:59,178 GROUP, THAT WAS QUERYING THE 1316 00:54:59,178 --> 00:55:02,348 ROLE OF INTERFERON ANTIBODIES 1317 00:55:02,348 --> 00:55:04,417 AND OTHER HOST-DIRECTED 1318 00:55:04,417 --> 00:55:07,487 AUTOANTIBODIES AND PERHAPS THEIR 1319 00:55:07,487 --> 00:55:10,223 CONTRIBUTION TO SEVERE COVID. 1320 00:55:10,223 --> 00:55:12,325 WE PROVIDED PLASMA SAMPLES TO 1321 00:55:12,325 --> 00:55:13,826 STEVE'S GROUP AND THEY QUERIED 1322 00:55:13,826 --> 00:55:17,997 THIS, WHAT I WOULD CALL A PILOT 1323 00:55:17,997 --> 00:55:20,233 STUDY, WAS NOT -- IT WAS NOT 1324 00:55:20,233 --> 00:55:22,802 SYSTEMATIC, DIDN'T INCLUDE ALL 1325 00:55:22,802 --> 00:55:24,103 OF OUR PATIENTS. 1326 00:55:24,103 --> 00:55:30,043 BUT THE RATES OF -- THE PRESENCE 1327 00:55:30,043 --> 00:55:31,878 OR QUANTIFICATION OF 1328 00:55:31,878 --> 00:55:32,579 HOST-DIRECTED AUTOANTIBODIES 1329 00:55:32,579 --> 00:55:34,180 DIDN'T MEANINGFULLY DIFFER IN 1330 00:55:34,180 --> 00:55:34,647 OUR COHORT -- 1331 00:55:34,647 --> 00:55:36,049 >> OTHER PATIENTS. 1332 00:55:36,049 --> 00:55:38,017 >> YEAH, TO THEIR OTHER 1333 00:55:38,017 --> 00:55:39,586 PATIENTS. 1334 00:55:39,586 --> 00:55:44,591 >> ANOTHER QUESTION ABOUT T 1335 00:55:44,591 --> 00:55:44,924 CELLS. 1336 00:55:44,924 --> 00:55:46,192 I THINK THE THINKING IS THAT 1337 00:55:46,192 --> 00:55:49,963 THERE'S A LOT OF 1338 00:55:49,963 --> 00:55:52,498 CROSS-REACTIVITY BEFORE WE WERE 1339 00:55:52,498 --> 00:55:55,101 EXPOSED TO COVID DUE TO 1340 00:55:55,101 --> 00:55:56,703 DIFFERENT COLD VIRUSES. 1341 00:55:56,703 --> 00:56:00,573 SO, HOW YOU WERE ABLE TO 1342 00:56:00,573 --> 00:56:03,977 DISTINGUISH AND SAY THESE ARE 1343 00:56:03,977 --> 00:56:05,378 REALLY COVID-SPECIFIC T CELLS, 1344 00:56:05,378 --> 00:56:09,415 DO YOU THINK THAT THEY WERE SOME 1345 00:56:09,415 --> 00:56:13,286 RAISED FROM PREVIOUS EXPOSURE TO 1346 00:56:13,286 --> 00:56:13,953 DIFFERENT VIRUSES AND 1347 00:56:13,953 --> 00:56:15,421 PATHOLOGICALLY IN A WAY -- 1348 00:56:15,421 --> 00:56:18,725 >> THAT'S TWO PARTS TO THE 1349 00:56:18,725 --> 00:56:19,826 QUESTION I'LL TRY TO ADDRESS. 1350 00:56:19,826 --> 00:56:22,695 ONE IS I THINK YOU'RE ABSOLUTELY 1351 00:56:22,695 --> 00:56:26,199 RIGHT. 1352 00:56:26,199 --> 00:56:26,666 THERE'S POTENTIAL FOR 1353 00:56:26,666 --> 00:56:28,434 CROSS-REACTIVE T-CELL RESPONSES 1354 00:56:28,434 --> 00:56:31,904 FROM EXPOSURE TO THE FOUR PRIOR 1355 00:56:31,904 --> 00:56:33,740 KNOWN ENDEMIC HUMAN 1356 00:56:33,740 --> 00:56:34,641 CORONAVIRUSES THAT DON'T INDUCE 1357 00:56:34,641 --> 00:56:35,508 SEVERE DISEASE. 1358 00:56:35,508 --> 00:56:37,777 AND MAYBE WE'RE SEEING SOME OF 1359 00:56:37,777 --> 00:56:38,578 THAT IN THE DIFFERENCES THAT 1360 00:56:38,578 --> 00:56:40,179 WE'RE SEEING AS A FUNCTION OF 1361 00:56:40,179 --> 00:56:40,446 AGE. 1362 00:56:40,446 --> 00:56:42,448 I MENTIONED TO YOU ABOUT OUR 1363 00:56:42,448 --> 00:56:43,683 T-CELL RESPONSES. 1364 00:56:43,683 --> 00:56:48,855 AS FAR AS THE ABILITY TO SAY ARE 1365 00:56:48,855 --> 00:56:51,824 THE INDIVIDUAL CLONOTYPES THAT 1366 00:56:51,824 --> 00:56:56,963 WE IDENTIFIED BY OUR TCR BETA 1367 00:56:56,963 --> 00:57:00,033 SEQUENCING COV SPECIFIC, TO THE 1368 00:57:00,033 --> 00:57:01,668 BEST OF MY KNOWLEDGE, YES BUT 1369 00:57:01,668 --> 00:57:06,272 THERE'S A CAVEAT, THE CAVEAT IS 1370 00:57:06,272 --> 00:57:06,673 THIS. 1371 00:57:06,673 --> 00:57:08,141 THAT ANSWER IS DEPENDENT UPON A 1372 00:57:08,141 --> 00:57:11,210 FUNCTIONAL ASSAY THAT WAS 1373 00:57:11,210 --> 00:57:17,684 PERFORMED BY ADAPTIVE 1374 00:57:17,684 --> 00:57:18,885 BIOTECHNOLOGIES USING 1375 00:57:18,885 --> 00:57:21,387 PROPRIETARY HIGH-THROUGHPUT 1376 00:57:21,387 --> 00:57:22,522 TECHNIQUE THAT DETERMINES 1377 00:57:22,522 --> 00:57:23,589 WHETHER OR NOT T CELLS, THESE 1378 00:57:23,589 --> 00:57:26,459 ARE T CELLS FOR THEIR ASSAY THAT 1379 00:57:26,459 --> 00:57:27,994 WORK FROM THE PBMC FRACTION, 1380 00:57:27,994 --> 00:57:28,194 RIGHT? 1381 00:57:28,194 --> 00:57:31,364 THOSE ARE THE ONES IMMEDIATELY 1382 00:57:31,364 --> 00:57:32,298 AVAILABLE. 1383 00:57:32,298 --> 00:57:34,867 WHETHER THEY ARE SPECIFIC 1384 00:57:34,867 --> 00:57:39,038 AGAINST, YOU KNOW, KNOWN COV 2 1385 00:57:39,038 --> 00:57:39,572 ANTIGENS. 1386 00:57:39,572 --> 00:57:41,007 NOW, I THINK THAT THE 1387 00:57:41,007 --> 00:57:43,042 SPECIFICITY OF THAT ASSAY IS 1388 00:57:43,042 --> 00:57:44,977 LIKELY GOOD, LIKE IN OTHER WORDS 1389 00:57:44,977 --> 00:57:47,613 IF THEY SAID THAT THE ASSAYS, 1390 00:57:47,613 --> 00:57:50,683 THIS IS COV 2 SPECIFIC AND WE 1391 00:57:50,683 --> 00:57:53,753 FOUND THE SAME TCR RECEPTOR THAT 1392 00:57:53,753 --> 00:57:56,622 RECOGNIZES WITH THE SAME MHC 1393 00:57:56,622 --> 00:57:58,024 CLASS YOU CAN GET CLOSE TO 1394 00:57:58,024 --> 00:58:00,226 SAYING OKAY MAYBE. 1395 00:58:00,226 --> 00:58:02,995 WHAT IS MISSING IN THE ASSAY IS 1396 00:58:02,995 --> 00:58:03,429 SENSITIVITY. 1397 00:58:03,429 --> 00:58:05,298 NOW, THEY CAN ONLY TEST SO MANY 1398 00:58:05,298 --> 00:58:07,600 T CELLS FUNCTIONALLY, RIGHT? 1399 00:58:07,600 --> 00:58:09,402 AND TIME HAS MOVED ON. 1400 00:58:09,402 --> 00:58:12,605 SO WHAT I'M CURIOUS ABOUT IN OUR 1401 00:58:12,605 --> 00:58:15,108 COHORT IS THE DENOMINATOR THAT I 1402 00:58:15,108 --> 00:58:18,644 SHOWED YOU IN THE COUPLE SLIDES, 1403 00:58:18,644 --> 00:58:23,816 THE NON-COV 2 SPECIFIC ANTIGEN, 1404 00:58:23,816 --> 00:58:31,691 T CELLS, SOME MAY MAY BE SPECIO 1405 00:58:31,691 --> 00:58:37,130 COS V 2, MAYBE GOING AGAINST HOT 1406 00:58:37,130 --> 00:58:37,997 OR OTHER PATHOGENS, BETTER 1407 00:58:37,997 --> 00:58:38,898 INSIGHT WOULD BE HELPFUL. 1408 00:58:38,898 --> 00:58:40,466 >> MAY BE AS IMPORTANT AS THE 1409 00:58:40,466 --> 00:58:41,267 COVID ONES. 1410 00:58:41,267 --> 00:58:42,168 >> A HUNDRED PERCENT. 1411 00:58:42,168 --> 00:58:42,769 >> THANK YOU. 1412 00:58:42,769 --> 00:58:45,204 >> SO WE'RE ABOUT OUT OF TIME 1413 00:58:45,204 --> 00:58:47,840 BUT I THINK NINA, DR. NINA 1414 00:58:47,840 --> 00:58:51,210 SCHOR, WOULD LIKE TO BRING US TO 1415 00:58:51,210 --> 00:58:52,011 A CLOSE. 1416 00:58:52,011 --> 00:58:53,479 >> HONESTLY, I JUST WANT TO 1417 00:58:53,479 --> 00:58:56,682 THANK YOU, DAN, FOR A FANTASTIC 1418 00:58:56,682 --> 00:59:00,319 DEMONSTRATION OF THE TRUE POWER 1419 00:59:00,319 --> 00:59:03,022 OF CLINICAL OBSERVATION AND 1420 00:59:03,022 --> 00:59:07,760 METICULOUS STUDY IN GENERATING 1421 00:59:07,760 --> 00:59:08,761 TESTABLE HYPOTHESES WITH 1422 00:59:08,761 --> 00:59:09,395 MECHANISTIC IMPORTANCE. 1423 00:59:09,395 --> 00:59:10,930 CONGRATULATIONS, AND THANK YOU. 1424 00:59:10,930 --> 00:59:13,933 >> THANK YOU SO MUCH, DR. SCHOR. 1425 00:59:13,933 --> 00:59:16,836 [APPLAUSE] 1426 00:59:16,836 --> 00:59:19,438 [END OF PROGRAM] 1427 00:59:19,438 --> 00:59:29,482