1 00:00:05,120 --> 00:00:06,888 GOOD AFTERNOON. WELCOME TO 2 00:00:06,888 --> 00:00:10,291 THE 26TH ANNUAL ASTUTE CLINICIAN 3 00:00:10,291 --> 00:00:14,262 LECTURE. I'M COLLEEN HAIGAN 4 00:00:14,262 --> 00:00:15,830 CHIEF MEDICAL OFFICER AND 5 00:00:15,830 --> 00:00:17,799 DIRECTOR AT THE NIH CLINICAL 6 00:00:17,799 --> 00:00:20,135 CERTAINTY. THE LECTURE IS NIH 7 00:00:20,135 --> 00:00:20,969 DIRECTOR'S WEDNESDAY AFTERNOON 8 00:00:20,969 --> 00:00:23,672 LECTURE SERIES, EVENT HOSTED BY 9 00:00:23,672 --> 00:00:24,739 THE CLINICAL CENTER. WE ARE 10 00:00:24,739 --> 00:00:26,307 PLEASED TO WELCOME BOTH IN 11 00:00:26,307 --> 00:00:28,643 PERSON ATTENDEES AND ALL OF OUR 12 00:00:28,643 --> 00:00:30,445 PARTICIPANTS WHO ARE WATCHING 13 00:00:30,445 --> 00:00:32,681 THROUGH THE NIH VIDEO WEBCAST. 14 00:00:32,681 --> 00:00:35,350 DR. TABAK UNFORTUNATELY WAS 15 00:00:35,350 --> 00:00:37,786 UNABLE TO JOIN US TODAY DUE TO 16 00:00:37,786 --> 00:00:40,755 OTHER COMMITMENTS. TODAY'S A 17 00:00:40,755 --> 00:00:44,993 LITTLE HOUSEKEEPING, TODAY'S CME 18 00:00:44,993 --> 00:00:49,197 CODE IS 50097. PLEASE TEXT THIS 19 00:00:49,197 --> 00:00:51,866 CODE TO THE JOHNS HOPKINS CME 20 00:00:51,866 --> 00:00:53,101 PHONE NUMBER ON THE SLIDE TO 21 00:00:53,101 --> 00:00:55,336 RECEIVE YOUR CME CREDITS FOR 22 00:00:55,336 --> 00:00:58,406 PARTICIPATION AT THIS LECTURE. 23 00:00:58,406 --> 00:00:59,874 FINALLY, LAST PIECE OF 24 00:00:59,874 --> 00:01:01,776 HOUSEKEEPING. DURING THE LECTURE 25 00:01:01,776 --> 00:01:02,877 IF YOU HAVE QUESTIONS PLEASE 26 00:01:02,877 --> 00:01:04,979 SUBMIT THEM FOR DR. SEARS ANY 27 00:01:04,979 --> 00:01:07,315 TIME DURING THE LECTURE BY 28 00:01:07,315 --> 00:01:08,750 CLICKING THE LIVE FEEDBACK 29 00:01:08,750 --> 00:01:10,785 BUTTON LOCATED ON THE VIDEOCAST 30 00:01:10,785 --> 00:01:12,987 WEBSITE. AND DR. SEARS WILL HAVE 31 00:01:12,987 --> 00:01:14,956 AN OPPORTUNITY TO ANSWER THOSE 32 00:01:14,956 --> 00:01:16,291 QUESTIONS AS TIME PERMITS AT THE 33 00:01:16,291 --> 00:01:18,326 CONCLUSION OF THE LECTURE. FOR 34 00:01:18,326 --> 00:01:19,561 INDIVIDUALS IN THE ROOM THEY CAN 35 00:01:19,561 --> 00:01:21,329 GO TO ONE OF THE MICS WHEN 36 00:01:21,329 --> 00:01:25,667 ASKING A QUESTION. SO FIRST, THE 37 00:01:25,667 --> 00:01:27,235 ASTUTE CLINICIAN LECTURESHIP IS 38 00:01:27,235 --> 00:01:29,137 ESTABLISHED THROUGH A GIFT FROM 39 00:01:29,137 --> 00:01:31,973 THE LATE ROBERT MILLER, AND HIS 40 00:01:31,973 --> 00:01:34,409 WIFE. IT HONORS A U.S. SCIENTIST 41 00:01:34,409 --> 00:01:36,511 WHO HAS OBSERVED AN UNUSUAL 42 00:01:36,511 --> 00:01:38,847 CLINICAL OCCURRENCE AND THEN BY 43 00:01:38,847 --> 00:01:40,849 INVESTIGATING IT HAS OPENED AN 44 00:01:40,849 --> 00:01:42,517 IMPORTANT NEW AVENUE OF 45 00:01:42,517 --> 00:01:44,652 RESEARCH. IT IS AN HONOR TO 46 00:01:44,652 --> 00:01:46,554 INTRODUCE CYNTHIA SEARS, AN 47 00:01:46,554 --> 00:01:48,757 INFECTIOUS DISEASE EXPERT IN GUT 48 00:01:48,757 --> 00:01:51,960 AND FOOD BORNE INFECTIONS. DR. 49 00:01:51,960 --> 00:01:53,561 SEARS' PRESENTATION TITLED 50 00:01:53,561 --> 00:01:58,066 SLEUTHING THE MICROBIOME, 51 00:01:58,066 --> 00:01:59,667 REVEALS UNDERCOVER AGENTS OF 52 00:01:59,667 --> 00:02:03,304 ONCOGENESIS. THIS WILL PROVIDE 53 00:02:03,304 --> 00:02:05,974 CONTEXT FOR MICROBE PROMOTION OF 54 00:02:05,974 --> 00:02:08,076 COLON CARCINOGENESIS AS 55 00:02:08,076 --> 00:02:10,712 ILLUSTRATED BY TWO VIGNETTES.SS 56 00:02:10,712 --> 00:02:13,815 ONE DEFINED BY A SINGLE 57 00:02:13,815 --> 00:02:14,816 BACTERIAL GENE AND SECOND 58 00:02:14,816 --> 00:02:17,652 COMMUNITY POTENTIAL TO MOLD 59 00:02:17,652 --> 00:02:20,922 DISEASE. SHE EARNED HER M.D. 60 00:02:20,922 --> 00:02:22,023 FROM THOMAS JEFFERSON MEDICAL 61 00:02:22,023 --> 00:02:23,658 COLLEGE AND COMPLETED RESIDENCY 62 00:02:23,658 --> 00:02:27,095 AND FELLOWSHIP AT NY HOSPITAL 63 00:02:27,095 --> 00:02:29,264 WEILL CORNELL MEDICAL CERTAINTY. 64 00:02:29,264 --> 00:02:30,732 IN 1985 DR. SEARS BEGAN HER 65 00:02:30,732 --> 00:02:32,600 TEACHING JOURNEY AS ASSISTANT 66 00:02:32,600 --> 00:02:34,035 PROFESSOR OF MEDICINE AT 67 00:02:34,035 --> 00:02:35,804 UNIVERSITY OF VIRGINIA BEFORE 68 00:02:35,804 --> 00:02:37,572 TRANSITIONING TO JOHNS HOPKINS 69 00:02:37,572 --> 00:02:38,506 UNIVERSITY SCHOOL OF MEDICINE 70 00:02:38,506 --> 00:02:41,075 WHERE SHE IS STILL TODAY. DR. 71 00:02:41,075 --> 00:02:44,479 SEARS IS A PHYSICIAN, MENTOR, 72 00:02:44,479 --> 00:02:46,014 AND ALSO PROFESSOR OF MEDICINE 73 00:02:46,014 --> 00:02:48,449 AND ONCOLOGY AT JOHNS HOPKINS 74 00:02:48,449 --> 00:02:49,150 UNIVERSITY SCHOOL OF MEDICINE 75 00:02:49,150 --> 00:02:51,920 AND PROFESSOR OF MOLECULAR 76 00:02:51,920 --> 00:02:53,087 MICROBIOLOGY AND IMMUNOLOGY AT 77 00:02:53,087 --> 00:02:57,826 THE HOPKINS BLOOMBERG SCHOOL OF 78 00:02:57,826 --> 00:03:01,596 PUBLIC HEALT HEALTH. SHE LEADS E 79 00:03:01,596 --> 00:03:02,831 MICROBIOME PROGRAM IF THE CANCER 80 00:03:02,831 --> 00:03:03,631 INSTITUTE FOR CANCER 81 00:03:03,631 --> 00:03:05,767 IMMUNOTHERAPY AT JOHNS HOPKINS 82 00:03:05,767 --> 00:03:06,935 AND DIRECTOR OF THE MURINE 83 00:03:06,935 --> 00:03:09,037 FACILITY AT JOHNS HOPKINS. THE 84 00:03:09,037 --> 00:03:10,839 LAB IS CONDUCTING HUMAN AND 85 00:03:10,839 --> 00:03:12,473 MURINE STUDIES TO DETERMINE IF 86 00:03:12,473 --> 00:03:14,242 AND HOW MICROBIOME IMPACTS 87 00:03:14,242 --> 00:03:16,678 CLINICAL RESPONSES TO CANCER 88 00:03:16,678 --> 00:03:18,146 IMMUNOTHERAPY P. AS INFECTIOUS 89 00:03:18,146 --> 00:03:19,914 DISEASE EXPERT DR. SEARS STUDIED 90 00:03:19,914 --> 00:03:22,217 GUT INFECTIONS INCLUDING 91 00:03:22,217 --> 00:03:23,585 DIARRHEAL AND FOOD BORNE 92 00:03:23,585 --> 00:03:29,858 ILLNESSES SUCH AS C DID C DIFF D 93 00:03:29,858 --> 00:03:31,092 HELICOBACTER. IN LABORATORY AND 94 00:03:31,092 --> 00:03:35,830 CLINICAL SETTINGS SHE STUDIED 95 00:03:35,830 --> 00:03:39,067 PATHOGENESIS FRAGILLIS OVER THE 96 00:03:39,067 --> 00:03:40,368 PAST 25 YEARS. THE CURRENT 97 00:03:40,368 --> 00:03:41,536 FOSSAS OF THE SEARS LABORATORY 98 00:03:41,536 --> 00:03:43,938 IS DETERMINE HOW THE MICROBIOTA 99 00:03:43,938 --> 00:03:47,575 AND SPECIFIC BACTERIAL PATHOGEN 100 00:03:47,575 --> 00:03:48,710 HEARSAY CONTRIBUTE TO COLON 101 00:03:48,710 --> 00:03:51,713 CARCINOGENESIS. THE LABORATORY 102 00:03:51,713 --> 00:03:53,748 INTEGRATES STUDIES OF HUMAN AND 103 00:03:53,748 --> 00:03:57,485 MOUSE MODELS, EMPLOYING 104 00:03:57,485 --> 00:04:00,488 MICROBIOLOGY, BIOINFORMATICS, 105 00:04:00,488 --> 00:04:01,756 IMMUNOLOGY, AND OTHER SCIENTIFIC 106 00:04:01,756 --> 00:04:04,425 METHODS, DR. SEARS HAS WORKED 107 00:04:04,425 --> 00:04:08,897 ABROAD IN THAILAND BRAZIL, 108 00:04:08,897 --> 00:04:09,797 HAITI, BANGLADESH AND MAY LA 109 00:04:09,797 --> 00:04:11,966 SHAH. SHE ALSO CERTAINED AS THE 110 00:04:11,966 --> 00:04:14,035 EDITOR OF THE ASSOCIATE AD E OF 111 00:04:14,035 --> 00:04:17,472 ED SR. EDITOR OF JOURNAL 112 00:04:17,472 --> 00:04:19,073 INFECTIOUS DISEASE BETWEEN 2000 113 00:04:19,073 --> 00:04:21,209 AND 2016 APPOINTED EDITOR IN 114 00:04:21,209 --> 00:04:23,411 CHIEF OF JOURNAL OF INFECTIOUS 115 00:04:23,411 --> 00:04:24,946 DISEASE IN JANUARY OF 2022. 116 00:04:24,946 --> 00:04:26,214 SHE'S MEMBER OF THE E TERM 117 00:04:26,214 --> 00:04:27,849 ADVISORY BOARD OF DIGESTIVE 118 00:04:27,849 --> 00:04:29,250 DISEASE RESEARCH CENTER AT 119 00:04:29,250 --> 00:04:31,085 VANDERBILT AND COLUMBIA 120 00:04:31,085 --> 00:04:32,120 UNIVERSITY, HAS BEEN AN ACTIVE 121 00:04:32,120 --> 00:04:34,155 MEMBER OF IDSA MORE THAN 20 122 00:04:34,155 --> 00:04:37,358 YEARS, SERVING THE SOCIETY IN 123 00:04:37,358 --> 00:04:38,626 NUMEROUS CAPACITY AND CURRENTLY 124 00:04:38,626 --> 00:04:40,662 PRESIDENT OF THAT ORGANIZATION. 125 00:04:40,662 --> 00:04:41,896 IN 2021 SHE WAS AWARDED THE 126 00:04:41,896 --> 00:04:43,231 JOHNS HOPKINS CANCER CENTER 127 00:04:43,231 --> 00:04:46,200 MENTORING AWARD. DR. SEARS IS 128 00:04:46,200 --> 00:04:47,802 RECOGNIZED GLOBALLY FOR HER 129 00:04:47,802 --> 00:04:50,338 SCIENTIFIC EXPERTISE AND IS A 130 00:04:50,338 --> 00:04:52,907 TRUE ROLE MODEL AS A CARING 131 00:04:52,907 --> 00:04:54,876 COMPASSIONATE CLINICIAN TO ALL 132 00:04:54,876 --> 00:04:57,879 PATIENTS. DR. SEARS EXEMPLIFIES 133 00:04:57,879 --> 00:04:59,881 THE TRAITS THAT FOR THE HONOR 134 00:04:59,881 --> 00:05:01,649 RES UPON THIS LECTURESHIP WHICH 135 00:05:01,649 --> 00:05:03,818 WAS CREATED 26 YEARS AGO. WITH 136 00:05:03,818 --> 00:05:04,986 NO FURTHER ADIEU, I WOULD LIKE 137 00:05:04,986 --> 00:05:06,521 TO INTRODUCE DR. SEARS. THANK 138 00:05:06,521 --> 00:05:06,721 YOU. 139 00:05:06,721 --> 00:05:13,728 [APPLAUSE] 140 00:05:13,728 --> 00:05:15,630 GOOD AFTERNOON, EVERYONE. I WILL 141 00:05:15,630 --> 00:05:16,531 CORRECT ONE THING I WAS 142 00:05:16,531 --> 00:05:20,969 PRESIDENT OF IDSA IN 2019 SO NOT 143 00:05:20,969 --> 00:05:23,271 THE CURRENT PRESIDENT, NOT TO 144 00:05:23,271 --> 00:05:25,873 TRY TO TAKE THAT HONOR TODAY. SO 145 00:05:25,873 --> 00:05:28,109 I WANT TO FIRST THANK DR. MILLER 146 00:05:28,109 --> 00:05:31,312 AND HITS FAMILY FOR SUPPORTING 147 00:05:31,312 --> 00:05:33,815 THIS LECTURE AND THEN DR. 148 00:05:33,815 --> 00:05:35,416 GILLMAN AND THE NIH COMMITTEE 149 00:05:35,416 --> 00:05:37,752 FOR SELECTING ME FOR THE HONOR 150 00:05:37,752 --> 00:05:40,989 OF SPEAKING TODAY AND TO ALL OF 151 00:05:40,989 --> 00:05:46,127 YOU, HERE IN -- AND ONLINE WHO 152 00:05:46,127 --> 00:05:48,629 JOINED TO LISTEN. SO THE TITLE 153 00:05:48,629 --> 00:05:50,465 AS YOU HAVE ALREADY HEARD IS 154 00:05:50,465 --> 00:05:53,234 SLEUTHING THE MICROBIOME REVEALS 155 00:05:53,234 --> 00:05:54,369 UNDERCOVER AGENTS OF 156 00:05:54,369 --> 00:06:01,142 ONCOGENESIS. AND FEW DISCLOSE 157 00:06:01,142 --> 00:06:03,378 SURES NOT PARTICULARLY DOMAIN. I 158 00:06:03,378 --> 00:06:05,747 HOPE TO GIVE A FRAMEWORK HOW WE 159 00:06:05,747 --> 00:06:07,715 CONSIDER MICROBES IN THE 160 00:06:07,715 --> 00:06:09,584 PATHOGENESIS OF HUMAN COLON 161 00:06:09,584 --> 00:06:12,020 CANCER, AND TALK TO YOU A LITTLE 162 00:06:12,020 --> 00:06:15,323 BIT ABOUT SPECIFIC BACTERIA AS 163 00:06:15,323 --> 00:06:16,891 WELL AS COMMUNITIES THAT MAYBE 164 00:06:16,891 --> 00:06:20,862 CONTRIBUTING TO THE DISEASE. SO 165 00:06:20,862 --> 00:06:24,532 THE OVERARCHING HYPOTHESIS IS 166 00:06:24,532 --> 00:06:26,167 THE GUT MICROBIOME IS A CRITICAL 167 00:06:26,167 --> 00:06:27,702 ENVIRONMENTAL FACTOR IMPACTING 168 00:06:27,702 --> 00:06:28,803 DISEASE AND THERAPEUTIC 169 00:06:28,803 --> 00:06:30,772 OUTCOMES. THAT CERTAINLY IS NOT 170 00:06:30,772 --> 00:06:33,207 UNIQUE TO OUR GROUP. BUT IF WE 171 00:06:33,207 --> 00:06:34,876 STEP BACK TOP THE FIELD OF 172 00:06:34,876 --> 00:06:38,112 INFECTIOUS DISEASES, I THINK WE 173 00:06:38,112 --> 00:06:39,680 NOW MUST CONSIDER THAT 174 00:06:39,680 --> 00:06:42,250 ANTIBIOTIC EXPOSURE OVER LIFE 175 00:06:42,250 --> 00:06:44,919 THROUGH ITS MICROBIOME IMPACTS 176 00:06:44,919 --> 00:06:47,355 IS AN EVER-EVOLVING DISEASE 177 00:06:47,355 --> 00:06:49,123 MODIFIER POTENTIALLY WITH HIGH 178 00:06:49,123 --> 00:06:52,193 IMPACT ON CHRONIC DISEASE 179 00:06:52,193 --> 00:06:54,128 DEVELOPMENT. SO AS COLLEEN JUST 180 00:06:54,128 --> 00:06:56,697 SAID I'M GOING TO PRESENT DATA 181 00:06:56,697 --> 00:06:59,267 TODAY WHERE WE TOGGLE BETWEEN 182 00:06:59,267 --> 00:07:03,137 HUMAN STUDIES AND MURINE STUDIES 183 00:07:03,137 --> 00:07:04,906 TRYING TO UNDERSTAND HOW THE 184 00:07:04,906 --> 00:07:08,643 MICROBIOME IS AFFECTING DISEASE. 185 00:07:08,643 --> 00:07:10,311 THIS IS THE BIG PICTURE FOR THE 186 00:07:10,311 --> 00:07:12,080 FIELD. ON ONE END OF THE 187 00:07:12,080 --> 00:07:15,083 SPECTRUM WE ARE CONSIDERING THE 188 00:07:15,083 --> 00:07:16,951 IMPACT OF SINGLE SPECIES. THE 189 00:07:16,951 --> 00:07:18,719 THREE THAT GET DISCUSSED MOST 190 00:07:18,719 --> 00:07:26,627 OFTEN ARE LISTED HERE, 191 00:07:26,627 --> 00:07:30,832 FUSOBACTERIUM, ESCHERICHIA 192 00:07:30,832 --> 00:07:34,068 CO-LIE AN INTERTOX GENIC 193 00:07:34,068 --> 00:07:36,037 BACTEROIDES FRAGILIS. WE CAN 194 00:07:36,037 --> 00:07:38,473 TAKE A MOUSE AND ADD ONE BUG TO 195 00:07:38,473 --> 00:07:41,109 ITS MICROBIOME AND LEAD TO 196 00:07:41,109 --> 00:07:43,644 COMPLETE CHANGE OF PHENOTYPE 197 00:07:43,644 --> 00:07:47,982 DEPENDING ON TOXIN IT MAKES AND 198 00:07:47,982 --> 00:07:49,183 IL 17 IN THE RIGHT GENETIC 199 00:07:49,183 --> 00:07:52,153 CONTEXT. THAT GENETIC CONTEXT IS 200 00:07:52,153 --> 00:07:54,922 A MOUSE THAT IS HETEROZYGOUS FOR 201 00:07:54,922 --> 00:07:58,893 THE GENE APC, WHICH STANDS FOR 202 00:07:58,893 --> 00:08:03,564 AD NO, MA'AMTUS POLYCOSIS COLI 203 00:08:03,564 --> 00:08:05,666 AND IS THE GENE HEREDITARY 204 00:08:05,666 --> 00:08:08,402 CANCER SYNDROME KNOWN AS 205 00:08:08,402 --> 00:08:13,374 FAMILIAL ADENOMITUS AND IT IS 206 00:08:13,374 --> 00:08:15,977 MUTATED OVER 80% OF SPORADIC 207 00:08:15,977 --> 00:08:17,979 COLON CANCER. EACH OF THE 208 00:08:17,979 --> 00:08:19,747 BACTERIA I JUST LISTED HAS 209 00:08:19,747 --> 00:08:21,682 SUBSTANTIVE CONNECTIONS, TO 210 00:08:21,682 --> 00:08:22,917 COLON CANCER IN HUMANS IN THE 211 00:08:22,917 --> 00:08:26,921 LITERATURE. ON THE OTHER SIDE OF 212 00:08:26,921 --> 00:08:29,290 THE EQUATION IS THE IDEA THAT 213 00:08:29,290 --> 00:08:31,592 MICROBIAL COMMUNITY IS IMPORTANT 214 00:08:31,592 --> 00:08:33,828 TO DISEASE DEVELOPMENT. AND IN 215 00:08:33,828 --> 00:08:35,730 THIS CARTOON WHAT I'M SHOWING 216 00:08:35,730 --> 00:08:37,932 YOU IS THE ENTIRE COLON IS 217 00:08:37,932 --> 00:08:40,668 COVERED WITH TWO LAYERS OF 218 00:08:40,668 --> 00:08:43,804 MUCOUS, ONE IS A LOOSE LAYER OF 219 00:08:43,804 --> 00:08:46,340 MUCOUS, READILY INVADED BY 220 00:08:46,340 --> 00:08:49,844 BACTERIA, THE SECOND IS THIS 221 00:08:49,844 --> 00:08:51,746 DENSE LAYER OF MUCOUS THAT 222 00:08:51,746 --> 00:08:53,881 STRETCHES THE AXIS OF THE COLON, 223 00:08:53,881 --> 00:08:55,783 LARGELY STERILE, EXCEPT WHEN THE 224 00:08:55,783 --> 00:08:57,585 RIGHT BUGS COME ALONG AND ARE 225 00:08:57,585 --> 00:08:59,921 ABLE TO PENETRATE THAT AND 226 00:08:59,921 --> 00:09:02,256 PIERCE IT AND THEN SIT RIGHT ON 227 00:09:02,256 --> 00:09:05,326 TOP OF THE EPITHELIAL CELLS. AND 228 00:09:05,326 --> 00:09:07,261 STUDYING THESE BIOFILMS HAS LED 229 00:09:07,261 --> 00:09:09,864 US TO CONCEPTS AND 230 00:09:09,864 --> 00:09:12,066 CARCINOGENESIS AS WELL AS 231 00:09:12,066 --> 00:09:14,735 SURPRISING POTENTIAL CONTRIBUTOR 232 00:09:14,735 --> 00:09:19,907 TO COLON CANCER. SO TWO 233 00:09:19,907 --> 00:09:21,676 VIGNETTES AS MENTIONED ALREADY 234 00:09:21,676 --> 00:09:24,745 TODAY. ONE IS THE POWER OF ONE, 235 00:09:24,745 --> 00:09:33,020 IT USES A MODEL ENTEROTOXIGENIC 236 00:09:33,020 --> 00:09:33,721 BACTEROIDES WHICH WE HAVE BEEN 237 00:09:33,721 --> 00:09:35,389 STUDYING A LONG TIME. THE SECOND 238 00:09:35,389 --> 00:09:37,992 IS POWER COMMUNITY, OBSERVATIONS 239 00:09:37,992 --> 00:09:40,094 OF BIOFILM, RETURN TO THE FIRST 240 00:09:40,094 --> 00:09:46,067 VIGNETTE, AND TALK ABOUT 241 00:09:46,067 --> 00:09:49,971 INTERTOXIGENIC BACTEROIDES 242 00:09:49,971 --> 00:09:53,174 FRAGILIS OR EBF. WHERE IN THE 243 00:09:53,174 --> 00:09:56,143 WORLD DOES ETBF COME FROM? THESE 244 00:09:56,143 --> 00:09:57,945 ARE KNOT NOT A HOUSEHOLD NAME, 245 00:09:57,945 --> 00:10:00,681 IN IN CLINICAL MEDICINE. AND IT 246 00:10:00,681 --> 00:10:04,218 TURNS OUT THESE ORGANISMS WERE 247 00:10:04,218 --> 00:10:05,586 DISCOVERED BY DR. A LITTLE 248 00:10:05,586 --> 00:10:06,988 MEYERS, VETERINARY SCIENTIST AT 249 00:10:06,988 --> 00:10:08,489 MONTANA STATE UNIVERSITY IN THE 250 00:10:08,489 --> 00:10:12,126 MID 1980s. AND HE TOOK ON THIS 251 00:10:12,126 --> 00:10:13,561 PERPLEXING PROBLEM OF DIARRHEAL 252 00:10:13,561 --> 00:10:15,463 DISEASE IN NEWBORN LAMBS WHICH 253 00:10:15,463 --> 00:10:17,865 APPEARED NOT TO FOLLOW THE USUAL 254 00:10:17,865 --> 00:10:20,368 PATH OF OTHER DIARRHEAL DISEASE. 255 00:10:20,368 --> 00:10:23,070 IN AN ELEGANT SERIES OF 256 00:10:23,070 --> 00:10:23,904 EXPERIMENTS TRYING TO FIGURE OUT 257 00:10:23,904 --> 00:10:25,539 WHAT WAS GOING ON AT THE END OF 258 00:10:25,539 --> 00:10:28,009 THE DAY, HE CAME TO THE 259 00:10:28,009 --> 00:10:30,344 CONCLUSION THAT IT WAS THIS 260 00:10:30,344 --> 00:10:32,647 BACTEROIDES FRAGILIS STRAIN IN 261 00:10:32,647 --> 00:10:34,115 THE LOOP STIMULATING THIS 262 00:10:34,115 --> 00:10:36,017 SECRETION. AND IN FACT, WHEN YOU 263 00:10:36,017 --> 00:10:38,119 PUT THE SUPERNATANTS FROM THAT 264 00:10:38,119 --> 00:10:41,589 BUG INTO ADDITIONAL LIGATED 265 00:10:41,589 --> 00:10:43,691 LOOPS THEY BLEW UP SO MUCH WITH 266 00:10:43,691 --> 00:10:44,592 FLUID THEY WOULD BURST SIMILAR 267 00:10:44,592 --> 00:10:47,028 TO WHAT YOU CAN SEE IN ALCOHOL 268 00:10:47,028 --> 00:10:56,037 CHOLERATOXIN. DR. BRAD SACK, ATS 269 00:10:56,037 --> 00:10:58,072 HOPKINS MEDICINE WAS A SLEUTH OF 270 00:10:58,072 --> 00:11:01,142 CHILDHOOD DIE REARIA. AT THAT 271 00:11:01,142 --> 00:11:04,245 TIME CHILDHOOD DIARRHEA WAS THE 272 00:11:04,245 --> 00:11:07,348 NUMBER ONE CAUSE OF MORBIDITY 273 00:11:07,348 --> 00:11:09,016 AND MORTALITY, ONLY HALF COULD 274 00:11:09,016 --> 00:11:11,786 BE EXPLAINED. SO HE NOTICED DR. 275 00:11:11,786 --> 00:11:14,188 MEYERS WORK AND TOOK IT TO THE 276 00:11:14,188 --> 00:11:17,058 FIELD WHERE HE WENT TO THE 277 00:11:17,058 --> 00:11:18,859 INDIAN HEALTH INDIAN HOST 278 00:11:18,859 --> 00:11:21,062 SERVICE HOSPITAL IN WHITE RIVER, 279 00:11:21,062 --> 00:11:23,898 ARIZONA AND DID EPIDEMIOLOGY 280 00:11:23,898 --> 00:11:30,237 STUDY LINKING ETBF TO HUMAN 281 00:11:30,237 --> 00:11:31,505 DIARRHEAL DISEASE. WHEN I FIRST 282 00:11:31,505 --> 00:11:33,507 WENT TO HOPKINS DR. SACK CALLED 283 00:11:33,507 --> 00:11:38,212 ME UP AND LITERALLY CAME TO MY 284 00:11:38,212 --> 00:11:41,048 OFFICE WITH THIS TUBE OF 20 FOLD 285 00:11:41,048 --> 00:11:42,516 CONCENTRATED CULTURE 286 00:11:42,516 --> 00:11:45,086 SUPERNATANTS FROM AN ETBF STRAIN 287 00:11:45,086 --> 00:11:48,356 THAT DR. MYERS SENT HIM AND SAID 288 00:11:48,356 --> 00:11:51,792 ALL THE ASSAYS WE TYPICALLY DO 289 00:11:51,792 --> 00:11:53,828 IN DIARRHEAL DISEASE TO DETECT 290 00:11:53,828 --> 00:11:56,597 TOXINS HAVE FAILED. I HEAR YOUR 291 00:11:56,597 --> 00:11:59,266 WORK WITH COLON EPITHELIAL 292 00:11:59,266 --> 00:12:00,034 CELLS, TAKE THIS AND SEE WHAT 293 00:12:00,034 --> 00:12:02,670 YOU CAN DO. SO FORTUNATELY WE 294 00:12:02,670 --> 00:12:05,506 WERE WORKING WITH THESE HUMAN 295 00:12:05,506 --> 00:12:10,144 COLON CANCER CELLS CALLED HT 29C 296 00:12:10,144 --> 00:12:11,712 1 CELLS. HAVING THIS TOOL YOU 297 00:12:11,712 --> 00:12:13,681 CAN SEE TOP THE CONTROL SELLS 298 00:12:13,681 --> 00:12:15,282 AND SEE WHAT HAPPENS AFTER YOU 299 00:12:15,282 --> 00:12:18,352 ADD IN YOUR PURIFIED BRFT OR THE 300 00:12:18,352 --> 00:12:19,487 CULTURE SUPERNATANT, WE 301 00:12:19,487 --> 00:12:21,655 CERTAINLY HAD A TOOL. AND WHICH 302 00:12:21,655 --> 00:12:24,458 IT ALLOWED US TO GO AND FIND THE 303 00:12:24,458 --> 00:12:26,827 BFT GENE AND PURIFIED THE TOXIN. 304 00:12:26,827 --> 00:12:28,996 LEARNING THAT IN FACT IT WAS 305 00:12:28,996 --> 00:12:31,065 ACTIVE AT VERY LOW 306 00:12:31,065 --> 00:12:33,434 CONCENTRATIONS. THIS IS AN ASSAY 307 00:12:33,434 --> 00:12:38,939 WE STILL USE TODAY. SO STEPPING 308 00:12:38,939 --> 00:12:41,809 FORWARD IT TURNS OUT ETBF IS 309 00:12:41,809 --> 00:12:44,245 CHARACTERIZED BY A SINGLE 310 00:12:44,245 --> 00:12:51,352 VIRULENCE FACTOR, BACTEROIDES 311 00:12:51,352 --> 00:12:53,654 FRAGILIS GENE, IT IS A TOXIN 312 00:12:53,654 --> 00:12:54,755 DEVELOPED BY THE ORGANISM AND 313 00:12:54,755 --> 00:12:56,490 MOLDED BY THE ORGANISM AND 314 00:12:56,490 --> 00:12:59,059 SECRETED AS A MATURE 315 00:12:59,059 --> 00:13:04,999 20-KILODALTON PROTEIN. IT IS A 316 00:13:04,999 --> 00:13:07,234 ZINC DEPENDENT PROTEASE TOXIN 317 00:13:07,234 --> 00:13:08,903 DUE TO A SINGLE CHROMOSOME GENE 318 00:13:08,903 --> 00:13:11,305 IN THE ORGANISM, OCCASIONAL LAY 319 00:13:11,305 --> 00:13:12,873 TWO COPIES. ON THE RIGHT SHOWS 320 00:13:12,873 --> 00:13:15,242 YOU THE SPECTRUM OF WHAT HAPPENS 321 00:13:15,242 --> 00:13:18,746 WITH BACTEROIDES FRAGILIS, CLOSE 322 00:13:18,746 --> 00:13:21,749 TO 90% OF US LISTENING ARE 323 00:13:21,749 --> 00:13:24,652 COLONIZED WITH B FRAG. AT ONE 324 00:13:24,652 --> 00:13:27,955 END ARE THE NON-TOX GENIC BACK 325 00:13:27,955 --> 00:13:30,958 TROY DEES FRAGILIS THOUGHT 326 00:13:30,958 --> 00:13:33,260 IMPORTANT TO MUCOSAL IMMUNE 327 00:13:33,260 --> 00:13:34,628 SYSTEM AND WORK DONE BY DR. 328 00:13:34,628 --> 00:13:36,330 CASPER AT HARVARD. THER END OF 329 00:13:36,330 --> 00:13:41,101 THE SPECTRUM ARE THESE B FRAG 330 00:13:41,101 --> 00:13:44,171 STRAINS PRO INFLAMMATORY AND 331 00:13:44,171 --> 00:13:46,240 TUMORIGENIC. IN HUMANS DATA HAS 332 00:13:46,240 --> 00:13:49,777 ALSO ACCRUED ASSOCIATING ETBF 333 00:13:49,777 --> 00:13:51,445 WITH INFLAMMATORY DIE RELEASE 334 00:13:51,445 --> 00:13:53,814 DISEASE AND INFLAMMATORY BOW WE 335 00:13:53,814 --> 00:14:01,155 WOULD DBOWELDISEASE AND COLON CS 336 00:14:01,155 --> 00:14:02,590 IS ONE SLIDE I SUMMARIZED BY 337 00:14:02,590 --> 00:14:03,958 LIFE AND MANY OTHERS ON ONE 338 00:14:03,958 --> 00:14:05,726 SLIDE SO LET ME WALK YOU THROUGH 339 00:14:05,726 --> 00:14:07,194 THIS. THE FIRST THING I WANT TO 340 00:14:07,194 --> 00:14:10,364 DO IS TO REVIEW THE TIME LINE. 341 00:14:10,364 --> 00:14:13,000 THIS IS NOW GETTING APC 342 00:14:13,000 --> 00:14:14,134 HETEROZYGOUS MOUSE WHICH WITH 343 00:14:14,134 --> 00:14:16,604 THE SINGLE INOCULATION TO THE 344 00:14:16,604 --> 00:14:18,506 MOUSE WE CAN GET COLONIZATION 345 00:14:18,506 --> 00:14:20,407 AND TURNS OUT WE GET INITIATION 346 00:14:20,407 --> 00:14:23,177 OF TUMOR FORMATION QUICKLY SO 347 00:14:23,177 --> 00:14:25,412 THAT BY ONE WEEK YOU CAN SEE 348 00:14:25,412 --> 00:14:28,148 THESE MICROSCOPIC TUMORS IN THE 349 00:14:28,148 --> 00:14:30,251 COLON MUCOSA OF THE MOUSE DUE TO 350 00:14:30,251 --> 00:14:32,720 LOSS OF HETERO ZYGOSITY. AND OF 351 00:14:32,720 --> 00:14:34,955 COURSE OVER TIME THEY GROW UNTIL 352 00:14:34,955 --> 00:14:37,558 THEY ARE BIGGER TUMORS. I WANT 353 00:14:37,558 --> 00:14:40,594 TO POINT OUT HOW IMPORTANT THESE 354 00:14:40,594 --> 00:14:44,632 STUDIES DR. LU AND AND DR. 355 00:14:44,632 --> 00:14:45,299 (INDISCERNIBLE) WHERE THEY 356 00:14:45,299 --> 00:14:47,134 WALKED WITH ME THE LAST 25 YEARS 357 00:14:47,134 --> 00:14:49,036 IN DEVELOPING THESE DATA. SO THE 358 00:14:49,036 --> 00:14:51,205 FIRST THING I WANT TO TELL YOU 359 00:14:51,205 --> 00:14:53,607 IS ABOUT THE TOXIN, THEN INDEED 360 00:14:53,607 --> 00:14:56,210 I MENTIONED THIS IS THE 361 00:14:56,210 --> 00:14:58,345 VIRULENCE FACTOR OF NOTE. HERE 362 00:14:58,345 --> 00:15:00,014 IS -- SEE IF I CAN DO IT. THERE 363 00:15:00,014 --> 00:15:01,248 WE GO. HERE IS THE EVIDENCE FOR 364 00:15:01,248 --> 00:15:03,284 THAT. IF E WE TAKE THE ORGANISM 365 00:15:03,284 --> 00:15:06,387 AND DO AND IN FRAME DELETION 366 00:15:06,387 --> 00:15:07,688 MUTANT SO THE TOXIN GENE IS NO 367 00:15:07,688 --> 00:15:10,491 LONGER THERE WE OBLITERATE ALL 368 00:15:10,491 --> 00:15:12,493 THAT TUMORIGENESIS AND 369 00:15:12,493 --> 00:15:13,694 INFLAMMATION. ONLY WHEN THAT 370 00:15:13,694 --> 00:15:15,296 GENE IS THERE DO YOU SEE ANY OF 371 00:15:15,296 --> 00:15:18,732 THESE BIOLOGICAL EFFECTS IN THE 372 00:15:18,732 --> 00:15:21,835 MICE. NOW, THE NEXT STEP IN THIS 373 00:15:21,835 --> 00:15:28,542 PROCESS THAT WE STUDIED WAS E 374 00:15:28,542 --> 00:15:30,210 CAD HEREIN CLEAVAGE, ACTIVATION 375 00:15:30,210 --> 00:15:32,046 OF CMIC AND CELL PROLIFERATION. 376 00:15:32,046 --> 00:15:33,614 HOW DID THAT COME TO POE BE? 377 00:15:33,614 --> 00:15:35,015 EARLY IN THE 1990 ET CETERA I 378 00:15:35,015 --> 00:15:36,050 WAS PRESENTING AT A CONFERENCE 379 00:15:36,050 --> 00:15:38,252 AND AS I WALKED AWAY FROM THE 380 00:15:38,252 --> 00:15:41,655 PODIUM DR. JAMES NELSON CHAIR OF 381 00:15:41,655 --> 00:15:43,324 THE DEPARTMENT OF CELL BIOLOGY 382 00:15:43,324 --> 00:15:45,693 AT STANFORD STOPPED ME AND SAID, 383 00:15:45,693 --> 00:15:47,561 YOU KNOW, THERE IS SOMETHING 384 00:15:47,561 --> 00:15:48,729 ABOUT THOSE CELLS THAT MAKES ME 385 00:15:48,729 --> 00:15:52,166 THINK OF E CAD HEREIN. I HAVE 386 00:15:52,166 --> 00:15:54,301 THREE AGENTS, WHY DON'T I SEND 387 00:15:54,301 --> 00:15:55,336 THEM TO YOU AND SEE WHAT YOU 388 00:15:55,336 --> 00:15:57,771 THINK. SO INDEED HE DID AND THIS 389 00:15:57,771 --> 00:16:00,240 WESTERN BLOT SHOWS THAT IN FACT 390 00:16:00,240 --> 00:16:04,111 DR. NELSON WAS SPOT ON. SO IN 391 00:16:04,111 --> 00:16:07,047 THE FIRST LANE IS BLOT SHOWING E 392 00:16:07,047 --> 00:16:09,783 CAD HEREIN, 120-KILODALTON 393 00:16:09,783 --> 00:16:16,523 PROTEIN THAT IS KEY TO THE 394 00:16:16,523 --> 00:16:18,726 COLONIC ENTHELIAL CELLS 395 00:16:18,726 --> 00:16:20,327 TOGETHER. LANES 2 AND FORWARD 396 00:16:20,327 --> 00:16:21,695 SHOW WHAT HAPPENS AFTER ADDITION 397 00:16:21,695 --> 00:16:24,264 OF BFT TO THE HD 29 CELLS AND 398 00:16:24,264 --> 00:16:26,166 WHAT YOU SEE IS THIS RAPID 399 00:16:26,166 --> 00:16:28,636 CLEAVAGE OVER 30 TO 45 MINUTES 400 00:16:28,636 --> 00:16:31,005 OF THE E CAD HEREIN AND WHAT THE 401 00:16:31,005 --> 00:16:33,941 BLOT HERE SHOWING RESIDUAL 402 00:16:33,941 --> 00:16:36,276 INTRACELLULAR DOMAIN. WHEN E CAD 403 00:16:36,276 --> 00:16:44,852 HEREIN GETS CLEAVED IT RELEASES 404 00:16:44,852 --> 00:16:47,054 IT RELEASES THIS EXTRA FRAGMENT 405 00:16:47,054 --> 00:16:48,022 WHICH I WILL SHOW YOU IN A 406 00:16:48,022 --> 00:16:55,362 MOMENT. THERE IS MY CARTOON. SO 407 00:16:55,362 --> 00:16:58,799 WHAT HAPPENED NEXT WAS TWO 408 00:16:58,799 --> 00:17:05,105 THINGS. PART ONE BURT VOGELSTEIN 409 00:17:05,105 --> 00:17:09,076 AND KEN KENSLER PUBLISHED 410 00:17:09,076 --> 00:17:10,878 SIMILAR PAPERS SAYING THE WINT 411 00:17:10,878 --> 00:17:12,179 PATHWAY IS CRITICAL TO THE 412 00:17:12,179 --> 00:17:14,048 DEVELOPMENT OF COLON CANCER. 413 00:17:14,048 --> 00:17:15,616 SIMULTANEOUSLY I DID EXPERIMENT 414 00:17:15,616 --> 00:17:17,751 I FORGOT TO STOP AND CAME BACK 415 00:17:17,751 --> 00:17:19,753 IN THE NEXT MORNING AND LOOKED 416 00:17:19,753 --> 00:17:21,855 IN THE INCUBATOR AND LOW AND 417 00:17:21,855 --> 00:17:24,324 BEHOLD THE TISSUE CULTURE FLASK 418 00:17:24,324 --> 00:17:27,227 WAS BRIGHT YELLOW. SO I THOUGHT 419 00:17:27,227 --> 00:17:28,562 I NOT ONLY MESS UP THE 420 00:17:28,562 --> 00:17:30,264 EXPERIMENT I CONTAMINATED THE 421 00:17:30,264 --> 00:17:33,133 FLASK. WHEN I LOOKED UNDER THE 422 00:17:33,133 --> 00:17:35,035 MICROSCOPE, IN FACT THE MENU WAS 423 00:17:35,035 --> 00:17:36,270 CLEAR. WHAT HAPPENED IS OVER 424 00:17:36,270 --> 00:17:38,706 NIGHT THE CELLS HAD PROLIFERATED 425 00:17:38,706 --> 00:17:41,175 MADLY AND HAD USED UP ALL THE 426 00:17:41,175 --> 00:17:44,611 GLUCOSE IN THE MEDIA. SO WE HAD 427 00:17:44,611 --> 00:17:46,246 NEVER DONE LONGER EXPERIMENTS, 428 00:17:46,246 --> 00:17:48,182 IT WAS A NEW OBSERVATION. SO 429 00:17:48,182 --> 00:17:51,385 HERE IS A CARTOON OF WHAT WE 430 00:17:51,385 --> 00:17:53,654 THINK HAPPENED. SO I MENTIONED E 431 00:17:53,654 --> 00:17:57,057 CADHERIN IS A STRUCTURAL PROTEIN 432 00:17:57,057 --> 00:18:00,394 IN THE PART OF THE JUNCTIONAL 433 00:18:00,394 --> 00:18:01,595 COMPLEX IN THE COLON AND 434 00:18:01,595 --> 00:18:03,263 TETHERED TO THE INTRACELLULAR 435 00:18:03,263 --> 00:18:05,265 DOMAIN OF E CADHERIN IS A LITTLE 436 00:18:05,265 --> 00:18:07,201 POOL OF BETA CATENIN. WHICH AT 437 00:18:07,201 --> 00:18:10,204 THE TIME WAS THOUGHT TO BE 438 00:18:10,204 --> 00:18:12,206 BIOLOGICALLY INERT. HOWEVER WE 439 00:18:12,206 --> 00:18:14,775 KNEW BFT BOUND TO A SPECIFIC 440 00:18:14,775 --> 00:18:16,076 RECEPTOR, WE DIDN'T KNOW WHAT IT 441 00:18:16,076 --> 00:18:18,612 WAS ON THE CELL, WE SHOWED THAT 442 00:18:18,612 --> 00:18:20,748 E CADHERIN GOT CLEAVED AND IN 443 00:18:20,748 --> 00:18:23,517 FACT WE WENT ON TO SHOW THAT 444 00:18:23,517 --> 00:18:25,452 BETA CATENIN TRANSLOCATED TO THE 445 00:18:25,452 --> 00:18:28,322 NUCLEUS ACTIVATED CMIC AND LED 446 00:18:28,322 --> 00:18:33,227 TO CELL PROLIFERATION. AS WE 447 00:18:33,227 --> 00:18:34,661 CONTINUED FORWARD TRYING TO 448 00:18:34,661 --> 00:18:36,930 UNDERSTAND THIS PROCESS, IN THE 449 00:18:36,930 --> 00:18:40,934 MOUSE WE LEARNED THAT THE TOXIC 450 00:18:40,934 --> 00:18:43,704 DNA DAMAGE SHOWN HERE BY GAMMA H 451 00:18:43,704 --> 00:18:45,706 2A STAINING WHICH IS A MEASURE 452 00:18:45,706 --> 00:18:48,642 OF DOUBLE STRANDED DNA BREAKS 453 00:18:48,642 --> 00:18:52,446 INDICATIVE OF DNA DAMAGE. WITH 454 00:18:52,446 --> 00:18:53,881 (INDISCERNIBLE) HERE IN THE 455 00:18:53,881 --> 00:18:56,016 AUDIENCE TODAY WE STUDIED 456 00:18:56,016 --> 00:18:57,284 ADDITIONAL TRANSDUCTION PATHWAYS 457 00:18:57,284 --> 00:19:01,622 AND IDENTIFIED THAT P STAT 3 WAS 458 00:19:01,622 --> 00:19:04,491 ACTIVATED AMONG ALL THE STATS 459 00:19:04,491 --> 00:19:05,726 AND THAT ACTIVATION 460 00:19:05,726 --> 00:19:07,194 PHOSPHORYLATION THAT PROTEIN IS 461 00:19:07,194 --> 00:19:09,563 CRITICAL TO THE PRODUCTION IL 17 462 00:19:09,563 --> 00:19:12,766 WHICH PROVED TO BE PRO 463 00:19:12,766 --> 00:19:17,204 CARCINOGENIC IN THIS MODEL. 464 00:19:17,204 --> 00:19:18,438 FRANK AND HIS TEAM HELPED 465 00:19:18,438 --> 00:19:19,473 FURTHER PULL THIS APART AND WHAT 466 00:19:19,473 --> 00:19:22,309 WE ULTIMATELY LEARNED IS THAT 467 00:19:22,309 --> 00:19:25,779 THE IL 17 RECEPTOR ON THE 468 00:19:25,779 --> 00:19:26,713 COLOLLIC EPITHELIAL CELL IS 469 00:19:26,713 --> 00:19:29,516 CRITICAL TO THE CARCINOGENESIS. 470 00:19:29,516 --> 00:19:31,885 ONCE THAT RESELL TORR IS ENGAGED 471 00:19:31,885 --> 00:19:35,389 BY IL 17 THERE IS NF KAPPA B 472 00:19:35,389 --> 00:19:36,857 ACTIVATION, RELEASED FROM THE 473 00:19:36,857 --> 00:19:40,027 EPITHELIAL CELLS, CXCL 474 00:19:40,027 --> 00:19:41,428 CHEMOKINES THEN THAT PULLS IN 475 00:19:41,428 --> 00:19:44,498 ALL THESE IMMATURE MYLOAD CELLS 476 00:19:44,498 --> 00:19:46,266 JUST TO THE DISTAL COLON WHERE 477 00:19:46,266 --> 00:19:49,269 WE SEE THE TUMORS THAT IS SO 478 00:19:49,269 --> 00:19:51,972 IMPORTANT TO THE TUMORIGENESIS 479 00:19:51,972 --> 00:19:56,276 THAT THE PICTURES SHOW. WE TRIED 480 00:19:56,276 --> 00:19:59,680 FOR YEARS TO FIGURE OUT BFT 481 00:19:59,680 --> 00:20:02,082 RECEPTOR WAS AND THROUGH AN SI 482 00:20:02,082 --> 00:20:05,953 RNA LIBRARY WE ZEROED IN ON GPR 483 00:20:05,953 --> 00:20:10,157 35 GTB BINDING PROTEIN THAT IS 484 00:20:10,157 --> 00:20:12,025 PREVALENT AND HAS HIGH 485 00:20:12,025 --> 00:20:14,228 EXPRESSION LEVEL IN THE GUT. 486 00:20:14,228 --> 00:20:15,929 HOWEVER, THAT WAS NOT THE 487 00:20:15,929 --> 00:20:17,798 RECEPTOR. WHEN YOU KNOCK OUT 488 00:20:17,798 --> 00:20:19,600 THAT PARTICULAR PROTEIN YOU GET 489 00:20:19,600 --> 00:20:24,137 MORE TUMORS NOT LESS TUMORS. SO 490 00:20:24,137 --> 00:20:27,074 IT REMAINED A PUZZLE WHAT THE 491 00:20:27,074 --> 00:20:33,480 BFT TOXIN WAS BINDING TO. ENTER 492 00:20:33,480 --> 00:20:37,150 MAX WHITE AN MST TALENTED MSTP 493 00:20:37,150 --> 00:20:38,619 STUDENT AT JOHNS HOPKINS. FROM 494 00:20:38,619 --> 00:20:42,456 THIS FIRST TIME I MET MAX, HOE 495 00:20:42,456 --> 00:20:44,658 TOLD HIM ABOUT THIS PROBLEM, HE 496 00:20:44,658 --> 00:20:46,059 GOT FASCINATED BY IT AND I WAS 497 00:20:46,059 --> 00:20:48,128 FORTUNATE TO HAVE HIM JOIN THE 498 00:20:48,128 --> 00:20:51,231 LAB. HE POSITED THAT BFT NEEDED 499 00:20:51,231 --> 00:20:53,734 HELP FROM SOME COMPONENT OF AN 500 00:20:53,734 --> 00:20:56,003 INTACT CELL TO CLEAVE E 501 00:20:56,003 --> 00:20:58,171 CADHERIN. SO IN TODAY'S WORLD HE 502 00:20:58,171 --> 00:21:00,207 TOOK ON A DIFFERENT TYPE OF 503 00:21:00,207 --> 00:21:00,974 GENETIC SCREEN AND IN THIS CASE 504 00:21:00,974 --> 00:21:05,846 IT WAS A CRISPER CAS 9 LIBRARY 505 00:21:05,846 --> 00:21:08,815 IN HT 9 CELLS, TURNED OUT THAT 506 00:21:08,815 --> 00:21:12,352 MATT WILDER AT HARVARD HAD 507 00:21:12,352 --> 00:21:16,456 ALREADY MADE THIS LIBRA LIBRARYT 508 00:21:16,456 --> 00:21:17,758 IS A DIE RELEASE DISEASE 509 00:21:17,758 --> 00:21:19,026 INVESTIGATOR. WE CALLED HIM AND 510 00:21:19,026 --> 00:21:21,795 HE INVITED MAX TO HIS LAB WHO 511 00:21:21,795 --> 00:21:23,297 WORKED WITH (INDISCERNIBLE) TO 512 00:21:23,297 --> 00:21:26,266 TEST THE LIBRARY. SO IN THE 513 00:21:26,266 --> 00:21:28,268 FIRST RUN THE ASSAY USED TO 514 00:21:28,268 --> 00:21:31,104 SCREEN THE LIBRARY WAS LOOKING 515 00:21:31,104 --> 00:21:34,441 FOR LOSS OF E CADHERIN. WHAT MAX 516 00:21:34,441 --> 00:21:37,711 CHOSE TO DO IS TO SORT OUT THE 517 00:21:37,711 --> 00:21:39,413 TOP 10 PERCENT OF CELLS THAT 518 00:21:39,413 --> 00:21:43,083 WERE STILL EXPRESSING E CADHERIN 519 00:21:43,083 --> 00:21:45,686 THINKING THAT IN FACT THOSE 520 00:21:45,686 --> 00:21:46,753 CELLS MUST HAVE LOST A GENE 521 00:21:46,753 --> 00:21:48,755 THROUGH THE LIBRARY THAT WAS 522 00:21:48,755 --> 00:21:53,160 CRITICAL TO THE ACTION OF BFT. 523 00:21:53,160 --> 00:21:54,761 THEN ON DEEP SEQUENCING THOSE 524 00:21:54,761 --> 00:21:56,897 CELLS THAT VERY FIRST RUN, ONE 525 00:21:56,897 --> 00:22:01,868 GENE POPPED OUT AS SIGNIFICANT 526 00:22:01,868 --> 00:22:03,403 AND THAT WAS (INAUDIBLE) 4 H. 527 00:22:03,403 --> 00:22:05,005 MAX WENT BACK AND REPEATED THE 528 00:22:05,005 --> 00:22:06,773 SCREEN AND SECOND TIME THROUGH 529 00:22:06,773 --> 00:22:10,477 ADDITION SAL GENES CAME UP BUT 530 00:22:10,477 --> 00:22:11,912 CLOUD AND 4 BECAME FURTHER 531 00:22:11,912 --> 00:22:13,613 ENRICHED. NOW I WILL SHOW YOU A 532 00:22:13,613 --> 00:22:16,483 SUBSET OF THE THE EXPERIMENTS 533 00:22:16,483 --> 00:22:21,655 THAT MAX HAS DONE TO TRY TO 534 00:22:21,655 --> 00:22:23,790 UNDERSTAND CLOUD AND 4 MAYBE A 535 00:22:23,790 --> 00:22:27,627 RECEPTOR FOR THE BFT PROTEIN P. 536 00:22:27,627 --> 00:22:31,198 SO IN THE FIRST FIGURE HERE WHAT 537 00:22:31,198 --> 00:22:34,868 I AM SHOWING YOU IS A CLAUDIN 4 538 00:22:34,868 --> 00:22:39,740 KNOCK OUT CELLS, HEK 29 CELLS 539 00:22:39,740 --> 00:22:42,442 LOSE STABLE BFT BINDING. IN THE 540 00:22:42,442 --> 00:22:44,678 BLOTS HERE YOU CAN SEE IN THE 541 00:22:44,678 --> 00:22:48,715 WILD TYPE CELLS YOU CAN DETECT 542 00:22:48,715 --> 00:22:51,952 CLAUDIN 4 AND BINDING OF BFT. IF 543 00:22:51,952 --> 00:22:55,188 YOU KNOCK OUT CLAUDIN 4 IN THOSE 544 00:22:55,188 --> 00:22:56,890 CELLS YOU LOSE THE SIGNAL. IF 545 00:22:56,890 --> 00:23:00,093 YOU KNOCK OUT CLAUDIN 3 AS 546 00:23:00,093 --> 00:23:01,161 POSSIBLE CONTROL THOSE SIGNALS 547 00:23:01,161 --> 00:23:04,131 ARE STILL THERE BECAUSE CLAUDIN 548 00:23:04,131 --> 00:23:06,867 4 IS STILL THERE. IN THE FLOW 549 00:23:06,867 --> 00:23:10,404 PLOTS YOU CAN SEE WHEN BFT BINDS 550 00:23:10,404 --> 00:23:11,738 CELLS IT SHIFTS CELLS TO THE 551 00:23:11,738 --> 00:23:13,306 RIGHT BECAUSE THIS PROTEIN HAS 552 00:23:13,306 --> 00:23:15,308 BEEN@ED TO THE CELLS. IF YOU USE 553 00:23:15,308 --> 00:23:16,443 -- ADDED TO THE CELLS. IF YOU 554 00:23:16,443 --> 00:23:19,880 USE THE C THE CLAUDIN 4 KNOCK 555 00:23:19,880 --> 00:23:22,182 OUT CELLS DON'T MOVE BECAUSE THE 556 00:23:22,182 --> 00:23:23,550 PROTEIN WON'T BIND. IF YOU USE 557 00:23:23,550 --> 00:23:26,219 THE CLAUDIN 3 KNOCK OUT CELLS 558 00:23:26,219 --> 00:23:28,221 THEY MOVE SIMILAR TO WILD TYPE. 559 00:23:28,221 --> 00:23:31,024 THE SECOND TYPE OF PROTOCOL MAX 560 00:23:31,024 --> 00:23:34,795 USED WAS TO TEST IF THE TOXIN 561 00:23:34,795 --> 00:23:37,631 WOULD CO-IMMUNOPRECIPITATE WITH 562 00:23:37,631 --> 00:23:38,832 CLAUDIN 4. THAT IS WHAT THIS 563 00:23:38,832 --> 00:23:43,270 PLOT SHOWSS. IF U YOU USED AN 564 00:23:43,270 --> 00:23:46,506 ANTI-CLAUDIN 4 ANTIBODY, GOT THE 565 00:23:46,506 --> 00:23:48,275 CLAUDIN 4 AND BFT AT THE SAME 566 00:23:48,275 --> 00:23:51,645 TIME IF YOU USE ANTI-CLAUDIN 2 567 00:23:51,645 --> 00:23:53,447 ANTIBODY HE DIDN'T GET THE THE 568 00:23:53,447 --> 00:23:56,683 SIGNALS. THE NEXT PROTOCOL WAS 569 00:23:56,683 --> 00:23:59,586 HEK 293T CELLS. THESE HAS NO 570 00:23:59,586 --> 00:24:02,489 ENDOGENOUS E CADHERIN OR 571 00:24:02,489 --> 00:24:05,058 CLAUDIN, IS. SO WHAT HE DID, IS 572 00:24:05,058 --> 00:24:08,662 HE TRANSFECTED IN CLAUDIN 4 AND 573 00:24:08,662 --> 00:24:10,230 THIS PLOT SHOWS WHETHER THE 574 00:24:10,230 --> 00:24:13,333 PROTEIN CLAUDIN 4 WAS 575 00:24:13,333 --> 00:24:16,002 FLUORESCENTLY LABELED OR NOT, 576 00:24:16,002 --> 00:24:18,405 THAT IN FACT THAT RESTORED 577 00:24:18,405 --> 00:24:21,675 BINDING OF BFT TO THE CELLS 578 00:24:21,675 --> 00:24:25,479 WHEREAS TRANSFECTING IN CLAUDIN 579 00:24:25,479 --> 00:24:30,250 2 DIDN'T RESTORE THAT BINDING. 580 00:24:30,250 --> 00:24:32,385 LASTLY, HE PUT THE WHOLE SYSTEM 581 00:24:32,385 --> 00:24:36,723 TOGETHER. HERE HE TOOK THE HEK 582 00:24:36,723 --> 00:24:40,927 293P CELLS, AND HE LABELED E 583 00:24:40,927 --> 00:24:43,697 CADHERIN WITH M CHERRY AS A 584 00:24:43,697 --> 00:24:45,565 FLUORESCENT PROBE. AND 585 00:24:45,565 --> 00:24:48,535 TRANSFECTED THAT IN TO THESE 586 00:24:48,535 --> 00:24:50,871 CELLS. HERE SHOWS THE FLOW PLOT 587 00:24:50,871 --> 00:24:54,241 SHOWING YOU THE POSITION OF THE 588 00:24:54,241 --> 00:24:56,376 E CADHERIN ON THE SURFACE OF THE 589 00:24:56,376 --> 00:25:01,481 CELLS. IF IF HE ADDS BFT TO THAT 590 00:25:01,481 --> 00:25:02,582 JUST EXPERIENCING E CADHERIN, 591 00:25:02,582 --> 00:25:05,452 THAT SIGNAL DOESN'T MOVE AT ALL. 592 00:25:05,452 --> 00:25:09,990 IF YOU CO-TRANSFECTS IN THE 593 00:25:09,990 --> 00:25:11,458 CLAUDIN 4 HE IN FACT AND THEN 594 00:25:11,458 --> 00:25:14,127 ADDS BFT YOU CAN SEE THE SIGNAL 595 00:25:14,127 --> 00:25:16,496 MOVE BECAUSE THE E CADHERIN HAS 596 00:25:16,496 --> 00:25:19,533 NOW BEEN CLEAVED AND THERE IS 597 00:25:19,533 --> 00:25:21,101 THAT 80-KILODALTON EXTRA 598 00:25:21,101 --> 00:25:25,205 CELLULAR DOMAIN THAT'S BEEN 599 00:25:25,205 --> 00:25:27,841 RELEASED. SO WHAT HAPPENS NEXT? 600 00:25:27,841 --> 00:25:31,211 MAX IS WORKING ON CLAUDIN 4 601 00:25:31,211 --> 00:25:33,680 MUTATIONAL ANALYSES, AND IN THIS 602 00:25:33,680 --> 00:25:40,153 CASE THIS IS A OF CLAUDIN 4 A 603 00:25:40,153 --> 00:25:41,721 TRANSMEMBRANE PROTEIN. HE SO FAR 604 00:25:41,721 --> 00:25:45,392 HAS TAKEN OFF THIS C TERMINUS 605 00:25:45,392 --> 00:25:48,461 SIGNALING DOMAIN, THAT IS VERY 606 00:25:48,461 --> 00:25:52,832 LITTLE EFFECT ON THE BINDING OF 607 00:25:52,832 --> 00:25:54,834 THE BFT. WE THOUGHT THAT THE 608 00:25:54,834 --> 00:25:56,636 LARGER EXTRA CELLULAR DOMAIN 609 00:25:56,636 --> 00:25:57,637 WOULD PROBABLY BE VERY IMPORTANT 610 00:25:57,637 --> 00:26:00,273 TO THE BINDING OF BFT BUT IN 611 00:26:00,273 --> 00:26:03,510 FACT THAT IS NOT CORRECT EITHER. 612 00:26:03,510 --> 00:26:04,678 TURNS OUT THAT LITTLE EXTRA 613 00:26:04,678 --> 00:26:12,419 CELLULAR DOMAIN, THE SMALLER NUB 614 00:26:12,419 --> 00:26:15,088 IS CRITICAL TO THE BFT THOUGH WE 615 00:26:15,088 --> 00:26:18,858 SUSPECT THE TWO DOMAINS ARE 616 00:26:18,858 --> 00:26:19,893 INTERACTIVE. THE OTHER THING WE 617 00:26:19,893 --> 00:26:23,029 ARE DOING IS WE ARE DISCUSSING 618 00:26:23,029 --> 00:26:26,566 WITH DR. XAVIER RUTH AND 619 00:26:26,566 --> 00:26:32,105 COLLEAGUE WOOLRICH ECHART, 620 00:26:32,105 --> 00:26:35,275 INSTITUTE OF BARCELONA, MODELING 621 00:26:35,275 --> 00:26:37,477 SYSTEM. DR. GOMEZ RUTH IS THE 622 00:26:37,477 --> 00:26:39,412 ONLY ONE WHO CRYSTALIZED BFT 623 00:26:39,412 --> 00:26:43,416 EVER. IN THE WORLD. SO WE SET UP 624 00:26:43,416 --> 00:26:46,753 A ZOOM CALL AND HAD A WONDERFUL 625 00:26:46,753 --> 00:26:48,588 TIME WORKING THROUGH THIS 626 00:26:48,588 --> 00:26:49,756 PROBLEM WITH THEM AND IF YOU 627 00:26:49,756 --> 00:26:50,957 HAVE QUESTIONS ABOUT THAT, I 628 00:26:50,957 --> 00:26:52,525 WOULD BE HAPPY TO ANSWER THEM AT 629 00:26:52,525 --> 00:26:58,198 THE END. SO STEPPING BACK, MAX 630 00:26:58,198 --> 00:27:00,567 HAS WRITTEN A VERY NICE REVIEW 631 00:27:00,567 --> 00:27:03,503 THAT GOT PUBLISHED THIS MONTH IN 632 00:27:03,503 --> 00:27:05,505 NATURE REVIEW MICROBIOLOGY, THIS 633 00:27:05,505 --> 00:27:08,575 IS WHAT WE THINK IS THE LIST OF 634 00:27:08,575 --> 00:27:11,578 CREDIBLE BACTERIAL CONTRIBUTORS 635 00:27:11,578 --> 00:27:14,314 TO COLON CANCER PATHOGENESIS. 636 00:27:14,314 --> 00:27:16,816 THERE IS 11 OF THEM ON THIS 637 00:27:16,816 --> 00:27:22,088 LIST. AS WE LOOK AT THE LINKAGE 638 00:27:22,088 --> 00:27:27,494 FOR ALL BUT SALMONELLA, THE 639 00:27:27,494 --> 00:27:30,830 LINKAGE IS HUMAN LINKAGE 640 00:27:30,830 --> 00:27:31,731 EPIDEMIOLOGICAL DATA SUGGESTS 641 00:27:31,731 --> 00:27:33,300 THESE RELEVANT TO COLON CANCER. 642 00:27:33,300 --> 00:27:35,568 WHEN WE MOVE TO ANIMAL MODELS IT 643 00:27:35,568 --> 00:27:37,370 GETS SLIMMER BECAUSE MANY OF 644 00:27:37,370 --> 00:27:41,174 THESE ORGANISMS DON'T READILY 645 00:27:41,174 --> 00:27:42,809 COLONIZE MICE. . BUT YOU CAN 646 00:27:42,809 --> 00:27:45,612 SEE A FEW DO. SO THERE IS 647 00:27:45,612 --> 00:27:47,647 OPPORTUNITIES TO STUDY IN MORE 648 00:27:47,647 --> 00:27:49,783 DETAIL THAN MECHANISMS OF THOSE 649 00:27:49,783 --> 00:27:54,020 PARTICULAR ORGANISMS. SO NOW WE 650 00:27:54,020 --> 00:27:56,289 WILL TURN TO VIGNETTE 2. HERE WE 651 00:27:56,289 --> 00:27:57,857 WILL TALK ABOUT POWER OF 652 00:27:57,857 --> 00:28:01,227 COMMUNITY AND DISCOVERIES WITH 653 00:28:01,227 --> 00:28:03,196 BIOFILMS. SO GOING TO START WITH 654 00:28:03,196 --> 00:28:07,734 A CLINICAL STUDY THAT WE DID 655 00:28:07,734 --> 00:28:09,436 WORKING WITH (INDISCERNIBLE) AND 656 00:28:09,436 --> 00:28:10,704 THE QUESTION WE WERE ASKING THIS 657 00:28:10,704 --> 00:28:13,273 WAS PUBLISHED IN GUT, WAS IS 658 00:28:13,273 --> 00:28:16,409 THERE AN ASSOCIATION BETWEEN ANY 659 00:28:16,409 --> 00:28:19,245 ORAL ANTIBIOTIC USE AND THE 660 00:28:19,245 --> 00:28:22,549 DEVELOPMENT OF COLON CANCER? SO 661 00:28:22,549 --> 00:28:25,585 WE WERE FORTUNATE SNUFF TO GET 662 00:28:25,585 --> 00:28:28,421 FUNDING FROM DR. PAUL ATWATER 663 00:28:28,421 --> 00:28:29,856 WHO RUNS AN ENVIRONMENTAL 664 00:28:29,856 --> 00:28:31,458 PROGRAM AT HOPKINS AND ABLE TO 665 00:28:31,458 --> 00:28:32,692 BUY DATA FROM THE CLINICAL 666 00:28:32,692 --> 00:28:35,061 PRACTICE RESEARCH DATA WHICH HAS 667 00:28:35,061 --> 00:28:38,498 DATA FROM 1989 TO 2012 AT THE 668 00:28:38,498 --> 00:28:40,734 TIME WE DID THIS. SO 669 00:28:40,734 --> 00:28:43,403 LONGITUDINAL DATA. THIS IS THE 670 00:28:43,403 --> 00:28:45,305 LARGEST CLINICAL DATABASE IN THE 671 00:28:45,305 --> 00:28:47,607 WORLD. WORKING WITH THEIR 672 00:28:47,607 --> 00:28:51,878 SCIENTISTS, WE DID A MATCHED 673 00:28:51,878 --> 00:28:54,681 DESIGN WHERE THE CONTROLS AND 674 00:28:54,681 --> 00:28:55,782 CANCER, COLON CANCERS ARE 675 00:28:55,782 --> 00:28:57,851 MATCHED BY YEAR OF BIRTH GENDER 676 00:28:57,851 --> 00:29:00,553 PRACTICE SITE AND CRPD 677 00:29:00,553 --> 00:29:02,155 REGISTRATION DATE. SO WE HAD THE 678 00:29:02,155 --> 00:29:04,791 OPPORTUNITY TO LOOK AT ALMOST 679 00:29:04,791 --> 00:29:08,695 29,000 COLON -- COLORECTAL 680 00:29:08,695 --> 00:29:11,698 CANCER AND 138,000 CONTROLS. THE 681 00:29:11,698 --> 00:29:15,835 CORE RESULTS ARE SHOWN HERE. ON 682 00:29:15,835 --> 00:29:18,338 THE HORIZONTAL AXIS ARE TOTAL 683 00:29:18,338 --> 00:29:22,275 DAYS OF EXPOSURE TO ORAL 684 00:29:22,275 --> 00:29:23,943 ANTIBIOTICS. THE STRIKING AND 685 00:29:23,943 --> 00:29:28,047 UNEXPECTED FINDING WAS THAT BY 686 00:29:28,047 --> 00:29:32,919 DAY 15 TOTAL ANTIBIOTIC EXPOSURE 687 00:29:32,919 --> 00:29:34,621 OF ANTIBIOTIC IN DATABASE, THERE 688 00:29:34,621 --> 00:29:38,258 WAS INCREASE ASSOCIATION WITH 689 00:29:38,258 --> 00:29:44,097 THE INCIDENCE OF COLON CANCER. 690 00:29:44,097 --> 00:29:45,865 IN CONTRAST AT RECTAL CANCER, 691 00:29:45,865 --> 00:29:49,669 EXPOSURE TO ANTIBIOTICS LED TO 692 00:29:49,669 --> 00:29:52,138 DECREASE RISK THAT SEEMED TO 693 00:29:52,138 --> 00:29:53,940 LEVEL OUT ABOUT DAY 60. WHICH 694 00:29:53,940 --> 00:29:56,109 ARE ABLE TO TAKE SOME OF THOSE 695 00:29:56,109 --> 00:29:57,877 COLON CANCERS AND FIGURE OUT 696 00:29:57,877 --> 00:29:59,979 WHERE THEY WERE FROM THE 697 00:29:59,979 --> 00:30:01,481 DATABASE AND SOME WERE PROXIMAL 698 00:30:01,481 --> 00:30:04,751 AND SOME WERE DISTAL. YOU CAN 699 00:30:04,751 --> 00:30:08,087 SEE 100% OF THE SIGNAL FOR COLON 700 00:30:08,087 --> 00:30:09,789 CANCER TRACED TO THE PROXIMAL 701 00:30:09,789 --> 00:30:13,893 COLON WITH NO EFFECT. IN THE 702 00:30:13,893 --> 00:30:17,130 DISTAL COLON. ONE OTHER PIECE OF 703 00:30:17,130 --> 00:30:18,765 INFORMATION WE FOUND CONVINCING 704 00:30:18,765 --> 00:30:20,733 WAS THAT THIS ASSOCIATION WITH 705 00:30:20,733 --> 00:30:25,004 ORAL ANTIBIOTIC EXPOSURE ONLY 706 00:30:25,004 --> 00:30:27,073 MATTERED FOR ORAL ANTIBIOTICS 707 00:30:27,073 --> 00:30:29,742 TAKEN MORE THAN 10 YEARS 708 00:30:29,742 --> 00:30:31,878 PREVIOUSLY. SO THIS IS NOT 709 00:30:31,878 --> 00:30:33,213 CONFOUNDING BY INDICATION AND 710 00:30:33,213 --> 00:30:35,114 THAT IS THE MAGIC NUMBER USED IN 711 00:30:35,114 --> 00:30:37,050 THE FIELD FROM THE TIME YOU GET 712 00:30:37,050 --> 00:30:38,818 THE START OF COLON CANCER UNTIL 713 00:30:38,818 --> 00:30:42,822 IT BECOMES A POLYP AND 714 00:30:42,822 --> 00:30:45,291 GASTROINTERROLOGIST SEES IT. 715 00:30:45,291 --> 00:30:46,392 SUBSEQUENTLY ACTUALLY THE 716 00:30:46,392 --> 00:30:48,761 HARVARD NURSES HEALTH STUDY WAS 717 00:30:48,761 --> 00:30:50,096 PUBLISHED JUST BEFORE OURS AND 718 00:30:50,096 --> 00:30:51,998 SUBSEQUENT STUDY OUT OF SWEDEN 719 00:30:51,998 --> 00:30:54,000 ALSO VALIDATED THIS IDEA THAT 720 00:30:54,000 --> 00:30:57,437 THE ORAL ANTIBIOTIC EXPOSURE AND 721 00:30:57,437 --> 00:31:00,440 SUBSEQUENT COLON CANCER WOULD BE 722 00:31:00,440 --> 00:31:03,676 DISTANT IN TIME. SO FOR ME THIS 723 00:31:03,676 --> 00:31:06,546 HIGHLIGHTS HOW ANTIBIOTIC 724 00:31:06,546 --> 00:31:08,515 MEDIATED MICROBIOME MAY 725 00:31:08,515 --> 00:31:13,353 CONTRIBUTE TO CHRONIC DISEASE 726 00:31:13,353 --> 00:31:16,222 DEVELOPMENT. SO NOW LET ME TELL 727 00:31:16,222 --> 00:31:18,091 YOU MORE ABOUT BIOFILMS. SO THIS 728 00:31:18,091 --> 00:31:20,059 IS THE WORK OF CHRISTINE 729 00:31:20,059 --> 00:31:22,228 (INAUDIBLE) AN SUBSEQUENTLY 730 00:31:22,228 --> 00:31:25,698 JULIA DREWAS. IN THE TOP FIGURE 731 00:31:25,698 --> 00:31:27,867 HERE, THIS IS A HEALTHY COLON, 732 00:31:27,867 --> 00:31:30,770 THAT LIGHT BLUE STAIN YOU ARE 733 00:31:30,770 --> 00:31:32,772 SEEING IS THAT DENSE INNER 734 00:31:32,772 --> 00:31:34,607 MUCOUS LAYER ON THE RIGHT OF THE 735 00:31:34,607 --> 00:31:37,176 EPITHELIAL CELLS. TO DO THIS 736 00:31:37,176 --> 00:31:39,412 PROJECT, WE SORT OF ROCKED BACK 737 00:31:39,412 --> 00:31:41,548 AND SAID THERE IS IS A LOT OF 738 00:31:41,548 --> 00:31:42,882 BUGS IN THE GUT. MAYBE WE 739 00:31:42,882 --> 00:31:45,351 SHOULDN'T PUT ALL OUR MONEY IN 740 00:31:45,351 --> 00:31:47,720 ONE BUCKET. LET'S BACK OFF AND 741 00:31:47,720 --> 00:31:50,256 TRY TO LOOK BROADER WAY AT HOW 742 00:31:50,256 --> 00:31:52,559 BACTERIA MAY BE AFFECTING COLON 743 00:31:52,559 --> 00:31:54,694 CANCER, WE WENT TO THE PATHOLOGY 744 00:31:54,694 --> 00:31:56,829 SUITE AND GOT PIECES OF TUMOR 745 00:31:56,829 --> 00:31:58,631 FROM THE SURFACE AND WE GOT 746 00:31:58,631 --> 00:32:02,168 PIECES OF TUMOR FROM THE FAR 747 00:32:02,168 --> 00:32:03,870 DISTAL MARGIN OF SURGICAL 748 00:32:03,870 --> 00:32:05,305 RESECTION, USUALLY HALF THE 749 00:32:05,305 --> 00:32:06,706 COLON, AND BROUGHT THEM BACK O 750 00:32:06,706 --> 00:32:12,378 THE LAB. AND THEN WE HYBRIDIZED 751 00:32:12,378 --> 00:32:15,048 THEM WITH BACTERIAL R RNA PROBE 752 00:32:15,048 --> 00:32:16,816 TO DETECT ALL BACTERIA THAT WERE 753 00:32:16,816 --> 00:32:21,354 THERE. TO SUR SURPRISE WE CAN 754 00:32:21,354 --> 00:32:23,723 SEE THAT PINK, THAT PINK IS A 755 00:32:23,723 --> 00:32:25,725 MASS OF BACTERIA SITTING ON TOP 756 00:32:25,725 --> 00:32:28,428 OF THE COLON TUMOR AND ON TOP OF 757 00:32:28,428 --> 00:32:30,797 THE EPITHELIAL CELLS IN THE 758 00:32:30,797 --> 00:32:32,265 NUCLEI IN THE CELLS STAINED IN 759 00:32:32,265 --> 00:32:36,069 BLUE BY DAPI. IN ADDITION IF YOU 760 00:32:36,069 --> 00:32:38,104 TAKE THAT NORMAL FLANKING IT 761 00:32:38,104 --> 00:32:43,343 ALSO HAD THESE DENSE AGGREGATES 762 00:32:43,343 --> 00:32:44,711 INVASIVE BIOFILM PRESENT AS 763 00:32:44,711 --> 00:32:47,246 WELL. ALMOST ALWAYS IF THE TUMOR 764 00:32:47,246 --> 00:32:49,248 SHOWS A BIOFILM THE FLAGGING 765 00:32:49,248 --> 00:32:50,950 NORMAL WAY AWAY FROM THE TUMOR 766 00:32:50,950 --> 00:32:53,152 IS ALSO GOING TO SHOW THAT 767 00:32:53,152 --> 00:32:54,153 LEADING US TO BELIEVE THAT 768 00:32:54,153 --> 00:32:55,989 PROBABLY THOSE BACTERIA ARE ALL 769 00:32:55,989 --> 00:32:58,524 ALONG THE SWATH OF THE TISSUE. 770 00:32:58,524 --> 00:33:02,061 WE WANTED TO KNOW WHAT THOSE 771 00:33:02,061 --> 00:33:03,529 BACTERIA WERE. SO WE USED A 772 00:33:03,529 --> 00:33:05,765 VARIETY OF PROBES IN AND THE 773 00:33:05,765 --> 00:33:10,003 BOTTOM LINE IS ALL OF THESE 774 00:33:10,003 --> 00:33:11,504 BIOFILMS ARE POLYMICROBIAL. WHEN 775 00:33:11,504 --> 00:33:13,373 WE DID THIS STAINING, WHAT WE 776 00:33:13,373 --> 00:33:16,542 COULD SEE IS THAT THE BACTERIA 777 00:33:16,542 --> 00:33:18,611 INVADED TUMOR. SO EVERY TUMOR 778 00:33:18,611 --> 00:33:23,149 WITH POSITIVE BIOFILM HAS AN 779 00:33:23,149 --> 00:33:26,452 INTRATUMOR BACTERIAL POPULATION. 780 00:33:26,452 --> 00:33:30,890 THE NEXT IS TO GO TO COLONOSCOPY 781 00:33:30,890 --> 00:33:33,026 SUITE TO SEE PATIENTS AVERAGE 782 00:33:33,026 --> 00:33:34,827 RISK FOR COLON CANCER AND WE 783 00:33:34,827 --> 00:33:39,465 LEARN SUBSEQUENTLY 10 TO 15% 784 00:33:39,465 --> 00:33:41,901 INDIVIDUALS UNDERGOING 785 00:33:41,901 --> 00:33:44,937 COLONOSCOPY HAVE BIOFILMS BUT 786 00:33:44,937 --> 00:33:46,272 SUNNY AND POLYMICROBIAL SITTING 787 00:33:46,272 --> 00:33:50,877 ON TOP OF THE CELLS. ADDITION 788 00:33:50,877 --> 00:33:51,844 WORK OVER TIME WE CAME TO 789 00:33:51,844 --> 00:33:55,181 UNDERSTAND THIS WAS A SPECTRUM 790 00:33:55,181 --> 00:33:57,083 OF BACTERIA BUT WITH THREE 791 00:33:57,083 --> 00:34:00,119 FLAVORS IF YOU WILL. MOST ARE 792 00:34:00,119 --> 00:34:02,455 POLYMICROBIAL THAT SHOW 793 00:34:02,455 --> 00:34:05,024 PREDOMINANCE OF BACTEROIDES, 794 00:34:05,024 --> 00:34:07,360 THERE IS LARGE SUBSET THAT ALSO 795 00:34:07,360 --> 00:34:11,130 HAVE FUSO BACTERIUM BLOOMS SHOWN 796 00:34:11,130 --> 00:34:13,066 IN YELLOW AND TINY SUBSET THAT 797 00:34:13,066 --> 00:34:15,401 IS MADE UP OF A COMPLEX OF 798 00:34:15,401 --> 00:34:18,638 DIFFERENT PROTEOBACTERIA GRAM 799 00:34:18,638 --> 00:34:21,407 NEGATIVE RODS. I'M DISAPPOINTED 800 00:34:21,407 --> 00:34:24,877 TO SAY THAT IN FACT WE STILL 801 00:34:24,877 --> 00:34:26,045 DON'T KNOW WHO EXACTLY IS 802 00:34:26,045 --> 00:34:30,149 SITTING IN THOSE INVASIVE 803 00:34:30,149 --> 00:34:33,019 BIOFILMS BUT CAROLINE WENTZEL, 804 00:34:33,019 --> 00:34:36,189 CELL MOLECULAR GRADUATE 805 00:34:36,189 --> 00:34:37,623 PROFESSOR IS ANALYZING THE DATA 806 00:34:37,623 --> 00:34:40,893 NOW SO HOPEFULLY WE WILL KNOW 807 00:34:40,893 --> 00:34:42,595 SOON. WE WERE DOING THIS STUDY 808 00:34:42,595 --> 00:34:44,297 AT HOPKINS AND DECIDED TO SET UP 809 00:34:44,297 --> 00:34:46,332 A PARALLEL STUDY WITH SOME 810 00:34:46,332 --> 00:34:49,202 FUNDING IN MALAYSIA. SO IN THIS 811 00:34:49,202 --> 00:34:51,237 CARTOON ON THE LEFT OF THE 812 00:34:51,237 --> 00:34:54,307 COLON, ALL THE RED DOTS, 813 00:34:54,307 --> 00:34:55,274 REPRESENT BIOFILM POSITIVE 814 00:34:55,274 --> 00:34:58,711 TUMORS WE EXAMINE AND THE BLUE 815 00:34:58,711 --> 00:35:00,480 DOTS ARE BIOFILM NEGATIVE TUMORS 816 00:35:00,480 --> 00:35:01,447 WHAT YOU CAN SEE IN THE 817 00:35:01,447 --> 00:35:04,550 SUMMATION HERE IS THAT ALMOST 818 00:35:04,550 --> 00:35:07,487 90% OF RIGHT COLON TUMORS WILL 819 00:35:07,487 --> 00:35:10,156 HAVE BIOFILMS. REMEMBER 820 00:35:10,156 --> 00:35:12,391 ANTIBIOTIC SLIDE. IN CONTRAST 821 00:35:12,391 --> 00:35:14,327 ONLY ABOUT HALF OF THE TUMORS IN 822 00:35:14,327 --> 00:35:16,462 THE LEFT COLON ARE POSITIVE. 823 00:35:16,462 --> 00:35:19,365 WHEN YOU GO TO THE COLONOSCOPY 824 00:35:19,365 --> 00:35:22,735 SUITE, IT IS AGNOSTIC TO 825 00:35:22,735 --> 00:35:24,637 SCIENCE, 10 TO 15% SHOW 826 00:35:24,637 --> 00:35:25,938 BIOFILMS. IN DATA I DON'T HAVE 827 00:35:25,938 --> 00:35:27,540 TIME TO SHOW YOU BUT I WISH TO 828 00:35:27,540 --> 00:35:30,276 TELL YOU IS WE LEARNED THESE 829 00:35:30,276 --> 00:35:35,047 BIOFILMS CAN BE CARCINOGENICICI. 830 00:35:35,047 --> 00:35:37,250 THEY AREN'T ALWAYS BUT CAN BE 831 00:35:37,250 --> 00:35:39,318 AND WE CAN TELL THAT BECAUSE MT. 832 00:35:39,318 --> 00:35:41,487 HUMAN TISSUE WE SEE DECREASED 833 00:35:41,487 --> 00:35:42,755 BARRIER FUNCTION BY LOSS OF E 834 00:35:42,755 --> 00:35:44,323 CADHERIN AND WE SEE ACTIVATION 835 00:35:44,323 --> 00:35:47,026 OF CARCINOGENIC MUCOSAL AND 836 00:35:47,026 --> 00:35:49,061 COLONIC EPITHELIAL CELL PATHWAYS 837 00:35:49,061 --> 00:35:51,063 LIKE IL 6 AND STAT 3. WE SEE 838 00:35:51,063 --> 00:35:53,866 THAT THE BIOFILMS ARE ASSOCIATED 839 00:35:53,866 --> 00:35:56,502 WITH COLON EPITHELIAL CELL 840 00:35:56,502 --> 00:36:00,273 PROLIFERATION. THEN WHEN WE PUT 841 00:36:00,273 --> 00:36:03,042 TUMORS INTO MICE, FLURRIES OF 842 00:36:03,042 --> 00:36:04,477 TUMORS INTO MICE WE GET 843 00:36:04,477 --> 00:36:10,750 INDUCTION OF TUMORS IN THOSE 844 00:36:10,750 --> 00:36:12,685 MICE. (INDISCERNIBLE) TOOK ON 845 00:36:12,685 --> 00:36:13,619 THE DIFFICULT CHALLENGE TRYING 846 00:36:13,619 --> 00:36:16,656 TO FIGURE OUT IF BIOFILMS IN 847 00:36:16,656 --> 00:36:19,058 EACH PERSON WERE 848 00:36:19,058 --> 00:36:21,127 PROCARCINOGENNIC. IN HER INITIAL 849 00:36:21,127 --> 00:36:23,796 ALNAL SEIZE SHE TRIPPED UPON 850 00:36:23,796 --> 00:36:28,100 BASICALLY, A TUMOR, 3728T, 851 00:36:28,100 --> 00:36:31,604 69-YEAR-OLD MAN WITH STAGE 1 852 00:36:31,604 --> 00:36:32,972 CARCINOMA IN THE RIGHT COLON 853 00:36:32,972 --> 00:36:35,107 THAT IN FACT WAS PLOTTED HERE 854 00:36:35,107 --> 00:36:38,578 ARE NUMBERS OF TUMORS AND THIS 855 00:36:38,578 --> 00:36:41,480 PARTICULAR SLURRY WAS PRO 856 00:36:41,480 --> 00:36:42,648 CARCINOGENIC. HERE IS SHOWING 857 00:36:42,648 --> 00:36:46,285 THE TUMORS IN THE DISTAL GUT. 858 00:36:46,285 --> 00:36:49,589 SHE QUALITIERED IT TO FIGURE HOW 859 00:36:49,589 --> 00:36:51,624 MANY BUGS WERE THERE. TO SUR 860 00:36:51,624 --> 00:36:57,797 SURPRISE WE DIDN'T FIND BFRAG OR 861 00:36:57,797 --> 00:37:00,900 MUCOBACTERIUM. WE DID NOTICE 862 00:37:00,900 --> 00:37:03,903 THAT IT HAD A CLOSTRIDIUM 863 00:37:03,903 --> 00:37:05,705 DIFFICILE IN THERE. SO IF YOU 864 00:37:05,705 --> 00:37:08,107 DID -- SHE TESTED THIS ISOLATES 865 00:37:08,107 --> 00:37:12,078 NOW AND IN FACT THAT REPLICATED 866 00:37:12,078 --> 00:37:16,082 THE RESULT WITH THE SLURRY. HE 867 00:37:16,082 --> 00:37:17,183 DID THE OBVIOUS NEXT EXPERIMENT 868 00:37:17,183 --> 00:37:20,653 AND PULLED THE C DIPF OUT OF THE 869 00:37:20,653 --> 00:37:21,921 ISOLATE AND LOOKED AT WHAT 870 00:37:21,921 --> 00:37:23,756 HAPPENED AND INDEED WHAT YOU CAN 871 00:37:23,756 --> 00:37:27,593 SEE HERE IS THAT WITHOUT THE C 872 00:37:27,593 --> 00:37:29,128 DIFF, I CAN'T CONTROL MY 873 00:37:29,128 --> 00:37:31,998 POINTER. WITHOUT THE C DIFF 874 00:37:31,998 --> 00:37:33,532 ISOLATES DON'T LEAD TO TUMOR 875 00:37:33,532 --> 00:37:35,368 FORMATION. THEY ONLY LEAD TO 876 00:37:35,368 --> 00:37:36,736 TUMOR FORMATION IN THE PRESENCE 877 00:37:36,736 --> 00:37:40,306 OF THE C DIFF. THESE ARE 878 00:37:40,306 --> 00:37:42,308 ADDITIONAL EXPERIMENTS WE DID TO 879 00:37:42,308 --> 00:37:47,146 TRY TO CONFIRM THESE OBSERVA 880 00:37:47,146 --> 00:37:49,515 OBSERVATIONS. JULIA IDENTIFIED A 881 00:37:49,515 --> 00:37:51,918 SLURRY FROM A PATIENT THAT WAS 882 00:37:51,918 --> 00:37:53,953 NOT PARTICULARLY TUMORIGENIC. 883 00:37:53,953 --> 00:37:58,190 WHAT SHE DID WAS ADDED THIS C 884 00:37:58,190 --> 00:37:59,926 DIFF TO THAT PARTICULAR SLURRY, 885 00:37:59,926 --> 00:38:03,396 INDEED IT DID CONVERT IT TO BE 886 00:38:03,396 --> 00:38:05,765 CARCINOGENIC IN THE MOUSE MODEL, 887 00:38:05,765 --> 00:38:09,502 NUMBERS OF TUMORS PLOTTED HERE. 888 00:38:09,502 --> 00:38:13,639 JADA AT ASTRA ZENECA DOING AN 889 00:38:13,639 --> 00:38:15,074 ENTIRELY DIFFERENT PROJECT IN 890 00:38:15,074 --> 00:38:17,944 THE LAB ON BIOFILM NEGATIVE 891 00:38:17,944 --> 00:38:22,648 TUMORS HAD ONE SAMPLE, THAT LED 892 00:38:22,648 --> 00:38:25,885 TO A LARGE NUMBER OF TUMORS AS 893 00:38:25,885 --> 00:38:28,454 SHOWN HERE. SHE DECIDED BECAUSE 894 00:38:28,454 --> 00:38:31,123 OF WHAT JULIA OBSERVED IN THE 895 00:38:31,123 --> 00:38:33,793 LAB TO LOOK FOR C DIFF AND THAT 896 00:38:33,793 --> 00:38:37,496 TOO WAS C DIFFICILE POSITIVE. 897 00:38:37,496 --> 00:38:42,501 ANGELA CHEN WHO IS OBTAINING HER 898 00:38:42,501 --> 00:38:43,436 Ph.D. IN MOLECULAR 899 00:38:43,436 --> 00:38:44,437 MICROBIOLOGY IMMUNOLOGY GROUP 900 00:38:44,437 --> 00:38:46,839 TOOK ON THE CHALLENGE OF SEEING 901 00:38:46,839 --> 00:38:49,208 IF THE SYSTEM WORKED IN SPECIFIC 902 00:38:49,208 --> 00:38:51,077 PATHOGEN FREE MICE AND INDEED 903 00:38:51,077 --> 00:38:54,080 SHE ORDERED TOX GENIC STRAINS 904 00:38:54,080 --> 00:38:56,882 AND NON-TOX GENIC STRAINS OF C 905 00:38:56,882 --> 00:39:00,886 DIFF FROM ATCC. THE TOXOXI GENIC 906 00:39:00,886 --> 00:39:03,689 STRAINS INDUCED TUMORS THE 907 00:39:03,689 --> 00:39:09,128 NON-TOXI GENIC DID NOT. SO MANY 908 00:39:09,128 --> 00:39:13,833 OF YOU IN THIS ROOM AND ONLINE 909 00:39:13,833 --> 00:39:17,470 KNOW CLOSTRIDIUM BACTEROIDES 910 00:39:17,470 --> 00:39:20,573 DIFFICILE INFECTION IS A DISEASE 911 00:39:20,573 --> 00:39:22,508 PRECIPITATED BY ANTIBIOTIC 912 00:39:22,508 --> 00:39:25,044 EXPOSURE IN MICROBIAL SHIFTS. 913 00:39:25,044 --> 00:39:28,981 THIS IS A OBLIGATE AN AROBE AND 914 00:39:28,981 --> 00:39:31,717 IT PRODUCES TWO TOXIN, TOXIN A 915 00:39:31,717 --> 00:39:33,686 AND B BUT IT IS TOXIN B THOUGHT 916 00:39:33,686 --> 00:39:36,222 TO BE MOST RELEVANT TO HUMAN 917 00:39:36,222 --> 00:39:38,591 DISEASE. WE NOW KNOW IN IS NOW 918 00:39:38,591 --> 00:39:40,159 THE MOST COMMON HEALTHCARE 919 00:39:40,159 --> 00:39:42,328 ASSOCIATED INFECTION IN THE 920 00:39:42,328 --> 00:39:45,331 U.S., A LEADING CAUSE OF GASTROO 921 00:39:45,331 --> 00:39:47,133 ENTERITIS IN THE U.S. BUT WE 922 00:39:47,133 --> 00:39:49,135 DON'T KNOW MUCH OF ANYTHING 923 00:39:49,135 --> 00:39:51,904 ABOUT ITS PERSISTENCE IN THE 924 00:39:51,904 --> 00:39:57,977 GUT. WE KNOW C DIFF RECURS IN 925 00:39:57,977 --> 00:39:59,645 ACUTE INFECTION BUT NO DATA 926 00:39:59,645 --> 00:40:02,214 AFTER C DIFF HOW MANY PEOPLE 927 00:40:02,214 --> 00:40:04,116 HANG ON TO IT FOR A PROTRACTED 928 00:40:04,116 --> 00:40:09,955 PERIOD OF TIME. NOW, JEWEL YAP 929 00:40:09,955 --> 00:40:10,990 PUBLISHED HER RESULTS WITH 930 00:40:10,990 --> 00:40:13,526 ANGELA CHEN AND OTHERS IN CANCER 931 00:40:13,526 --> 00:40:16,228 DISCOVERY AND SHE PROVIDED 932 00:40:16,228 --> 00:40:17,730 EVIDENCE TO LINK FOUR MECHANISMS 933 00:40:17,730 --> 00:40:22,201 TO THE CARCINOGENICITY OF C 934 00:40:22,201 --> 00:40:24,570 DIFFICILE BUT TODAY I WILL ONLY 935 00:40:24,570 --> 00:40:26,305 SHOW YOU DATA ABOUT THE BIOFILMS 936 00:40:26,305 --> 00:40:29,408 AN LINKAGE TO TOXIN AND THEN I 937 00:40:29,408 --> 00:40:30,643 WILL SHOW YOU UNPUBLISHED DATA 938 00:40:30,643 --> 00:40:33,646 WORKING ON THE COMMUNITY WHICH I 939 00:40:33,646 --> 00:40:38,951 THINK IS VERY INTERESTING. SO 940 00:40:38,951 --> 00:40:40,386 HERE AGAIN, WE ARE BACK TO THE 941 00:40:40,386 --> 00:40:47,093 MOUSE. . WHAT JULIA DID SHE 942 00:40:47,093 --> 00:40:48,928 INOCULATED THE MICE TO ALL MICE 943 00:40:48,928 --> 00:40:52,131 BUT NOT C DIFF THE PINK IS 944 00:40:52,131 --> 00:40:54,233 MASSIVE BACTERIA IN THE LUMEN OF 945 00:40:54,233 --> 00:40:57,903 THE GUT AND THE BLACK SPACE 946 00:40:57,903 --> 00:40:59,939 BETWEEN PINK AND BLUE, NUCLEI OF 947 00:40:59,939 --> 00:41:03,642 THE EPITHELIAL CELLS, IS THAT 948 00:41:03,642 --> 00:41:05,945 BLACK SPACE, THAT IS AN INNER 949 00:41:05,945 --> 00:41:07,279 MUCOUS LAYER LARGELY NOT 950 00:41:07,279 --> 00:41:08,347 PENETRATED HERE THOUGH 951 00:41:08,347 --> 00:41:10,082 OCCASIONALLY IS, EVEN WITH THE 952 00:41:10,082 --> 00:41:11,717 ISOLATES AS YOU CAN SEE IN THIS 953 00:41:11,717 --> 00:41:13,953 PICTURE. IF YOU LOOK AT THIS 954 00:41:13,953 --> 00:41:18,457 PICTURE ON THE RIGHT WHICH IS 955 00:41:18,457 --> 00:41:20,392 INCREASED MAGNIFICATION YOU 956 00:41:20,392 --> 00:41:25,831 DON'T SEE BACTERIA IN THE CRIBS 957 00:41:25,831 --> 00:41:28,367 OF THE COLON EPITHELIUM. LOOK AT 958 00:41:28,367 --> 00:41:29,668 THE BOTTOM NOW, THIS IS THE 959 00:41:29,668 --> 00:41:32,771 ISOLATES INOCULATED WITH C DIFF 960 00:41:32,771 --> 00:41:35,107 AND THE STRUCTURE OF THE BIOFILM 961 00:41:35,107 --> 00:41:38,444 IS DRAMATICALLY DIFFERENT. IN 962 00:41:38,444 --> 00:41:39,912 ADDITION LOOK AT THE PHOTO ON 963 00:41:39,912 --> 00:41:43,449 THE FAR RIGHT, NOW THESE 964 00:41:43,449 --> 00:41:44,617 BACTERIA REACH DOWN INTO THE 965 00:41:44,617 --> 00:41:46,318 CRYPTS WHICH IS WHERE THE 966 00:41:46,318 --> 00:41:47,586 COLONIC STEM CELLS ARE WHICH IS 967 00:41:47,586 --> 00:41:50,122 THE PROGENITOR CELL FOR COLON 968 00:41:50,122 --> 00:41:51,323 CANCER. 969 00:41:51,323 --> 00:41:53,959 I THINK OVER HERE YOU CAN SEE IT 970 00:41:53,959 --> 00:41:56,128 THOSE YELLOW RODS, THOSE ARE THE 971 00:41:56,128 --> 00:41:58,030 C DIFF. SO IT IS NOT BECAUSE 972 00:41:58,030 --> 00:42:00,199 THERE IS AN OVERWHELMING AMOUNT 973 00:42:00,199 --> 00:42:02,101 OF C D I F F THERE, IT IS 974 00:42:02,101 --> 00:42:04,069 BECAUSE THEY CHANGED THE 975 00:42:04,069 --> 00:42:10,309 FUNCTION OF THE COMMUNITY. WITH 976 00:42:10,309 --> 00:42:14,713 THE HELP OF (INDISCERNIBLE) C 977 00:42:14,713 --> 00:42:16,448 DIFF IS VERY HARD THE MUTATE WE 978 00:42:16,448 --> 00:42:17,783 OBTAINED SOME STRAINS THAT HAD 979 00:42:17,783 --> 00:42:23,956 THE TOXIN B KNOCKED OUT. WHEN 980 00:42:23,956 --> 00:42:25,758 JEW LA AND I TREATED THE -- 981 00:42:25,758 --> 00:42:27,126 JULIA TREATED THE MICE, 982 00:42:27,126 --> 00:42:29,461 COLONIZED THE MICE WITH TOXIN B 983 00:42:29,461 --> 00:42:31,163 STRAINS THEY COLONIZED FINE BUT 984 00:42:31,163 --> 00:42:34,567 DIDN'T INDUCE TUMORS LIKE THE 985 00:42:34,567 --> 00:42:38,137 TOXIN B POSITIVE STRAINS. T SO C 986 00:42:38,137 --> 00:42:39,838 DIFFICILE POSITIVE COMMUNITY 987 00:42:39,838 --> 00:42:42,975 INDUCES TUMORS THAT AGAIN IS 988 00:42:42,975 --> 00:42:44,410 APPEARS TO BE DRIVEN BY 989 00:42:44,410 --> 00:42:46,212 VIRULENCE GENE OF C D I F F 990 00:42:46,212 --> 00:42:53,052 WHICH IS TOXIN B. NOW I WILL 991 00:42:53,052 --> 00:42:54,653 DESCRIBE WHAT WE ARE DOING 992 00:42:54,653 --> 00:43:00,459 TODAY, THE WORK OF HADLY POUR 993 00:43:00,459 --> 00:43:01,493 GUARD CELL AND MOLECULAR 994 00:43:01,493 --> 00:43:04,330 GRADUATE STUDENT, EARLIER WORK 995 00:43:04,330 --> 00:43:07,066 DONE BY REECE KNIPEL WHO IS A 996 00:43:07,066 --> 00:43:10,703 POST DOC WORKING IN THE LAB. SO 997 00:43:10,703 --> 00:43:11,971 WE GOT INTERESTED IN WHAT ARE 998 00:43:11,971 --> 00:43:14,206 THE REST OF THOSE ISOLATES DOING 999 00:43:14,206 --> 00:43:17,476 IN THAT BIOFILM? SO I WANTED TO 1000 00:43:17,476 --> 00:43:19,545 DESCRIBE THE TWO EXPERIMENTAL 1001 00:43:19,545 --> 00:43:20,145 PROTOCOLS THEN SHOW YOU THE 1002 00:43:20,145 --> 00:43:25,684 DATA. SO IN THE FIRST PROTOCOL 1003 00:43:25,684 --> 00:43:29,188 WE EXAMINE HOW CULTURE 1004 00:43:29,188 --> 00:43:30,356 SUPERNATANTS FROM SOME OF THE 1005 00:43:30,356 --> 00:43:35,461 BACTERIA IN THAT BIOFILM, MIGHT 1006 00:43:35,461 --> 00:43:37,429 INFLUENCE GROWTH OF C DIFFICILE, 1007 00:43:37,429 --> 00:43:41,133 DONE BY GROWING STRAINS FOR 72 1008 00:43:41,133 --> 00:43:43,435 HOURS FILTER STERILIZING THEM 1009 00:43:43,435 --> 00:43:44,303 GETTING RID OF THE BUG AND 1010 00:43:44,303 --> 00:43:46,739 PUTTING THE C D I F F BACK IN 1011 00:43:46,739 --> 00:43:48,741 AND SEEING IF IT GROWS OR NOT. 1012 00:43:48,741 --> 00:43:50,042 IN THE SECOND PROTOCOL, 1013 00:43:50,042 --> 00:43:52,144 BASICALLY DOING THE SAME THING 1014 00:43:52,144 --> 00:43:56,582 BUT USING THE CULTURE 1015 00:43:56,582 --> 00:43:58,117 SUPERNATANT AT 72 HOURS AFTER 1016 00:43:58,117 --> 00:44:00,753 THE C D I F F WASSALLED AND IT 1017 00:44:00,753 --> 00:44:02,588 GREW -- ADDED AND IT GREW TO SEE 1018 00:44:02,588 --> 00:44:04,323 HOW MUCH TOXIN PRODUCED BY THE C 1019 00:44:04,323 --> 00:44:07,059 D I F F ON THAT CULTURE. AND 1020 00:44:07,059 --> 00:44:08,894 OVER HERE ARE VERO CELLS, A 1021 00:44:08,894 --> 00:44:12,197 CLASSIC ASSAY FOR C DIFFICILE 1022 00:44:12,197 --> 00:44:14,433 TOXINS. ON THE TOP NORMAL CELLS, 1023 00:44:14,433 --> 00:44:17,903 ON THE BOTTOM THEY ROUND UP WHEN 1024 00:44:17,903 --> 00:44:20,606 THEY SEE TOXIN A OR B IT TURNS 1025 00:44:20,606 --> 00:44:28,514 OUT. REECE HAD DONE THIS 1026 00:44:28,514 --> 00:44:29,248 PHENOMENALLY COMPLICATED SET OF 1027 00:44:29,248 --> 00:44:31,283 EXPERIMENTS TO FIGURE OUT WHICH 1028 00:44:31,283 --> 00:44:35,120 BUGS IN THE 30 MEMBER COMMUNITY 1029 00:44:35,120 --> 00:44:36,255 WERE IMPORTANT TO INTERACTING 1030 00:44:36,255 --> 00:44:38,223 WITH C D I F F. ONE THAT POPPED 1031 00:44:38,223 --> 00:44:48,801 OUT FROM HIS WORK WAS C CELLAELA 1032 00:44:50,703 --> 00:44:53,939 PNEUMONIA. SO PUT D C I F F IN 1033 00:44:53,939 --> 00:44:55,174 FRESH MEDIUM IT GROWS PINE. IF 1034 00:44:55,174 --> 00:44:57,409 YOU TAKE MEDIA AT 72 HOURS 1035 00:44:57,409 --> 00:44:59,845 FILTER STERILIZE AND REINOCULATE 1036 00:44:59,845 --> 00:45:02,348 IT WITH C DIFF RED LINE ON THE 1037 00:45:02,348 --> 00:45:06,752 BOTTOM, IT DOESN'T GROW. AT ALL 1038 00:45:06,752 --> 00:45:09,355 HOWEVER TAKE 72 HOUR CULTURE 1039 00:45:09,355 --> 00:45:10,823 FILTRATES OF A RANGE OF GRAM 1040 00:45:10,823 --> 00:45:14,093 NEGATIVE RODS INCLUDING CLUB 1041 00:45:14,093 --> 00:45:16,962 SEAL LA PNEUMONIAA AND 1042 00:45:16,962 --> 00:45:19,765 REINOCULATE WITH C DIFF THEY 1043 00:45:19,765 --> 00:45:22,668 GROW WELL AS SHOWN HERE. WE 1044 00:45:22,668 --> 00:45:24,436 THINK OR KNOW THAT CULTURE 1045 00:45:24,436 --> 00:45:26,572 SUPERNATANTS AND PROTEOBACTERIA 1046 00:45:26,572 --> 00:45:31,577 IN FACT RESTORE C DIFFICILE 1047 00:45:31,577 --> 00:45:33,045 GROWTH. THIS IS THE EXPERIMENT 1048 00:45:33,045 --> 00:45:36,014 DONE ADDITIONAL EXPERIMENT DONE 1049 00:45:36,014 --> 00:45:39,017 BY REECE AT ASTRA ZENECA AND IN 1050 00:45:39,017 --> 00:45:40,786 THE ZERO COLUMN IS WHETHER OR 1051 00:45:40,786 --> 00:45:42,721 NOT HE TOOK ALL THOSE ISOLATES 1052 00:45:42,721 --> 00:45:45,190 GREW THEM UP FOR 72 HOURS, 1053 00:45:45,190 --> 00:45:47,993 FILTER STERILIZED THEM, GREW THE 1054 00:45:47,993 --> 00:45:53,799 C DIFF AND TESTED THE TOXIN 1055 00:45:53,799 --> 00:45:56,402 PRODUCTION IN THAT CULTURE SOUP. 1056 00:45:56,402 --> 00:45:59,104 WITHOUT ADDITION AND JUST AS C 1057 00:45:59,104 --> 00:46:00,806 DIFF ALONE THE LEVEL OF TOXIN 1058 00:46:00,806 --> 00:46:03,108 PRODUCTION YOU SEE, AS YOU ADD 1059 00:46:03,108 --> 00:46:04,576 PROGRESSIVELY MORE OF THE 1060 00:46:04,576 --> 00:46:08,447 CULTURE SUPERNATANT FROM THE 1061 00:46:08,447 --> 00:46:10,682 OTHER BUGS YOU IN FACT GET 1062 00:46:10,682 --> 00:46:15,587 REDUCTION IN TOXIN PRODUCTION. I 1063 00:46:15,587 --> 00:46:16,488 WILL TELL YOU WHERE YOU KNOW 1064 00:46:16,488 --> 00:46:22,194 MORE THING. WE HAVE NOW THIS 1065 00:46:22,194 --> 00:46:24,430 FACTOR IN CULTURE SUPERNATANTS 1066 00:46:24,430 --> 00:46:29,101 APPEARS LESS THAN 4-KILODALTONS, 1067 00:46:29,101 --> 00:46:31,804 ITS HEAT LYPASE AND PROTEASE 1068 00:46:31,804 --> 00:46:33,505 RESISTANT. SO WE ARE SUSPICIOUS 1069 00:46:33,505 --> 00:46:38,777 OF A SMALL MOLECULE METABOLITE 1070 00:46:38,777 --> 00:46:39,945 MEDIATING THESE EFFECTS. HERE 1071 00:46:39,945 --> 00:46:42,080 IS WORKING HYPOTHESIS, THAT YOU 1072 00:46:42,080 --> 00:46:45,217 TAKE C DIFF IT IS HELPER BUGS IN 1073 00:46:45,217 --> 00:46:48,086 THOSE BIOFILMS AND YOU PUTS 1074 00:46:48,086 --> 00:46:50,088 THEMSELVE TOGETHER AND GET LESS 1075 00:46:50,088 --> 00:46:55,694 TOXIN PRODUCTION BY C DIFF BUT 1076 00:46:55,694 --> 00:46:57,596 SUSTAIN THE GROWTH A PERFECTED 1077 00:46:57,596 --> 00:46:59,531 STORM. NAMELY THE HELPER STRAINS 1078 00:46:59,531 --> 00:47:02,267 IN THE COLONIC MICROBIOME 1079 00:47:02,267 --> 00:47:05,437 PREVENT GROWTH OF THE ORGANISM 1080 00:47:05,437 --> 00:47:06,038 BUDDY MINISH TOXIN PRODUCTION 1081 00:47:06,038 --> 00:47:08,907 WHICH ENABLES IT TO REMAIN AS A 1082 00:47:08,907 --> 00:47:11,577 PERMANENT OR A CHRONIC COLONIZER 1083 00:47:11,577 --> 00:47:13,378 OF THE HUMAN GUT LIMITING THE 1084 00:47:13,378 --> 00:47:17,349 AMOUNT OF INFLAMMATION INDUCED 1085 00:47:17,349 --> 00:47:18,984 AND WE KNOW INFLAMMATION IS 1086 00:47:18,984 --> 00:47:21,253 CRITICAL TO THE DEVELOPMENT OF 1087 00:47:21,253 --> 00:47:29,361 COLON CANCER. NEXT STEPS, DOES C 1088 00:47:29,361 --> 00:47:31,363 DIFF HAVE A PLAUSIBLE LINKAGE TO 1089 00:47:31,363 --> 00:47:33,031 COLON CANCER, IT IS A COMPLETELY 1090 00:47:33,031 --> 00:47:36,568 NEW TOPIC. WE STARTED TO LOOK. 1091 00:47:36,568 --> 00:47:39,104 IF WE GO TO MALAYSIA COHORT, WE 1092 00:47:39,104 --> 00:47:43,308 HAVE NOW SEQUENCED 110 COLON 1093 00:47:43,308 --> 00:47:46,178 CANCER FROM THAT COHORT AND 29 1094 00:47:46,178 --> 00:47:49,581 OF THE 110 ARE POSITIVE FOR C 1095 00:47:49,581 --> 00:47:51,049 DIFFICILE IN THE SEQUENCE 1096 00:47:51,049 --> 00:47:55,487 ANALYSES. 25 OF THE 29 OF THOSE 1097 00:47:55,487 --> 00:47:57,990 ARE IN FACT BIOFILM POSITIVE. 1098 00:47:57,990 --> 00:48:00,325 WHEN WE LOOK AT OUR HOPKINS 1099 00:48:00,325 --> 00:48:02,995 COLON CANCER COHORT, PLEATED 1100 00:48:02,995 --> 00:48:05,564 OVER TIMES AND ONLY LOOK AT SIX 1101 00:48:05,564 --> 00:48:08,133 COLON CANCER TO DATE, FIVE OF 1102 00:48:08,133 --> 00:48:13,572 SIX WERE POSITIVE FOR C DIFFER 1103 00:48:13,572 --> 00:48:20,345 SILL AND TWO TOXIJENIC ANOTHER 1104 00:48:20,345 --> 00:48:22,347 POSSIBLE THING TO THINK ABOUT IS 1105 00:48:22,347 --> 00:48:24,783 C DIFFICILE DISEASE INCREASED 1106 00:48:24,783 --> 00:48:27,286 OVER TIME AND WE HAVE NEW 1107 00:48:27,286 --> 00:48:28,787 EPIDEMIOLOGY IN COLON CANCER 1108 00:48:28,787 --> 00:48:31,924 WITH ONSET OF EARLY ONSET 1109 00:48:31,924 --> 00:48:33,625 COLORECTAL CANCER WHICH WE ARE 1110 00:48:33,625 --> 00:48:36,962 SEEING IN INDIVIDUALS UNDER AGE 1111 00:48:36,962 --> 00:48:39,298 50 BEFORE SCREENING COLONOSCOPY 1112 00:48:39,298 --> 00:48:41,867 IS TYPICALLY RECOMMENDED. 1113 00:48:41,867 --> 00:48:44,636 LASTLY WE KNOW INFLAMMATORY 1114 00:48:44,636 --> 00:48:46,672 BOWEL DISEASE THERE IS A TON OF 1115 00:48:46,672 --> 00:48:50,008 C DIFFICILE COLONIZATION AND/OR 1116 00:48:50,008 --> 00:48:52,711 DISEASE, THAT IS A CONDITION 1117 00:48:52,711 --> 00:48:57,249 THAT ALSO HAS INCREASED RISK OF 1118 00:48:57,249 --> 00:48:59,985 COLON CANCER DEVELOPMENT. SO 1119 00:48:59,985 --> 00:49:03,722 WHAT ARE WE ACTUALLY DOING? 1120 00:49:03,722 --> 00:49:05,290 CYNTHIA SAY FELL IN THE LAB AND 1121 00:49:05,290 --> 00:49:09,494 SEAN ANDERSON FELL FELLOW IN THE 1122 00:49:09,494 --> 00:49:11,063 LAB ARE REEXAMINING HOW OFTEN WE 1123 00:49:11,063 --> 00:49:15,901 ARE ALL EXPOSED TO C DIFFICILE 1124 00:49:15,901 --> 00:49:18,503 LOOKING AT IGG ANTIBODIES TO 1125 00:49:18,503 --> 00:49:22,374 TOXIN B. WE HAVE COLLECTED 400 1126 00:49:22,374 --> 00:49:25,844 SAMPLES OF SERA AT HOPKINS, 1127 00:49:25,844 --> 00:49:27,012 PEDIATRIC AND ADULT AND IN THE 1128 00:49:27,012 --> 00:49:27,913 PROCESS OF TRYING TO GET THAT 1129 00:49:27,913 --> 00:49:35,687 DONE. WE HAVE ALSO INITIATED A 1130 00:49:35,687 --> 00:49:36,922 LISTENING TUESDAYNAL 1131 00:49:36,922 --> 00:49:38,190 COLONIZATION STUDY, 12 MONTH 1132 00:49:38,190 --> 00:49:39,758 STUDY IN ADULTS HOPE TO EXTEND 1133 00:49:39,758 --> 00:49:42,327 TO OUTPATIENTS AND PEDIATRICS 1134 00:49:42,327 --> 00:49:46,932 AGAIN LED BY SEAN ANDERSON TO 1135 00:49:46,932 --> 00:49:49,534 GET INITIAL C DIFF ISOLATE THAT 1136 00:49:49,534 --> 00:49:51,103 OVER TIME POTENTIALLY HOW LONG 1137 00:49:51,103 --> 00:49:54,706 IS THE ORGANISM CARRIED, WHAT IS 1138 00:49:54,706 --> 00:49:57,109 THE COMMUNITY ASSOCIATED WITH. 1139 00:49:57,109 --> 00:49:59,144 LASTLY WE ARE TRYING TO QUERY 1140 00:49:59,144 --> 00:50:01,413 LARGE DATABASES HOPKINS AND 1141 00:50:01,413 --> 00:50:02,714 ULTIMATELY NETWORK HOSPITALS TO 1142 00:50:02,714 --> 00:50:05,784 TRY TO LOOK AT THE INTERSECTION 1143 00:50:05,784 --> 00:50:08,153 OF C DIFFICILE INFECTION 1144 00:50:08,153 --> 00:50:10,122 COLONOSCOPY AND COLORECTAL 1145 00:50:10,122 --> 00:50:12,457 CANCER DETECTION. THIS IS LED BY 1146 00:50:12,457 --> 00:50:16,328 AND ALL STAR GROUP IN THE ID 1147 00:50:16,328 --> 00:50:19,531 DIVISION, I AM MOSTLY OBSERVER 1148 00:50:19,531 --> 00:50:20,932 LEARNER HERE BUT SEAN IS WORKING 1149 00:50:20,932 --> 00:50:23,502 ON THIS WITH MATTHEW ROBINSON, 1150 00:50:23,502 --> 00:50:28,073 AND KELLY AT HOPKINS. WITH THAT 1151 00:50:28,073 --> 00:50:31,376 I WANT TO TURN TO SOME TRIBUTES 1152 00:50:31,376 --> 00:50:33,412 OBVIOUSLY I AM STANDING HERE 1153 00:50:33,412 --> 00:50:34,613 BECAUSE OF THE SUPPORT 1154 00:50:34,613 --> 00:50:37,349 MENTORSHIP I RECEIVED OVER TIME 1155 00:50:37,349 --> 00:50:39,384 SO I WANT TO THANK DICK GRANT 1156 00:50:39,384 --> 00:50:41,887 WHETHER OR NOT WAS MY FIRST 1157 00:50:41,887 --> 00:50:43,588 MENTOR AT UNIVERSITY OF VIRGINIA 1158 00:50:43,588 --> 00:50:45,490 WHO TOOK ME INTO HIS LAB DESPITE 1159 00:50:45,490 --> 00:50:48,393 FACT I HAD NO IDEA HOW TO 1160 00:50:48,393 --> 00:50:50,562 PIPETTE. AND JULIE IN THE 1161 00:50:50,562 --> 00:50:52,931 DEPARTMENT OF PHARMACOLOGY THERE 1162 00:50:52,931 --> 00:50:55,033 OBVIOUSLY -- I WAS FIRST M.D. 1163 00:50:55,033 --> 00:50:56,702 PERSON SHE LET WORK IN HER LAB 1164 00:50:56,702 --> 00:50:59,271 AND I LEARNED A TREMENDOUS 1165 00:50:59,271 --> 00:51:00,939 AMOUNT, WE ARE STILL FAST 1166 00:51:00,939 --> 00:51:02,507 FRIENDS. IN PARTICULAR I LEARNED 1167 00:51:02,507 --> 00:51:05,077 A LOT ABOUT WRITING AS SHE 1168 00:51:05,077 --> 00:51:07,012 DUBBED ME THE QUEEN OF THE 1169 00:51:07,012 --> 00:51:12,217 MISPLACED MODIFIER. AT JOHNS 1170 00:51:12,217 --> 00:51:16,388 HOPKINS I WAS CO-HIRED BY MARK 1171 00:51:16,388 --> 00:51:18,957 (INAUDIBLE) THEN HEAD OF PI AND 1172 00:51:18,957 --> 00:51:20,926 JOHN BARTLETT HEAD OF ID AND 1173 00:51:20,926 --> 00:51:22,661 RECEIVED I NOR MOUSE SUPPORT 1174 00:51:22,661 --> 00:51:23,962 FROM THEM OVER THE LAST DECADES 1175 00:51:23,962 --> 00:51:26,631 I TOLD YOU HOW INVALUABLE 1176 00:51:26,631 --> 00:51:29,301 GETTING TO KNOW DR. SACK WAS AND 1177 00:51:29,301 --> 00:51:31,937 LASTLY DREW PERDOLE NOW ALMOST 1178 00:51:31,937 --> 00:51:34,239 20 YEARS SINCE I TOTAL HIM I 1179 00:51:34,239 --> 00:51:36,575 DIDN'T KNOW ANYTHING ABOUT THE 1180 00:51:36,575 --> 00:51:38,176 MUCOSAL IMMUNE SYSTEM AND COULD 1181 00:51:38,176 --> 00:51:40,045 HE HELP ME OUT AND INDEED HE 1182 00:51:40,045 --> 00:51:41,747 HAS, WE REMAIN WONDERFUL 1183 00:51:41,747 --> 00:51:46,685 COLLABORATORS. I ALSO WANT TO 1184 00:51:46,685 --> 00:51:49,387 REALLY THANK (INDISCERNIBLE) WU 1185 00:51:49,387 --> 00:51:51,022 AND (INDISCERNIBLE) THERE WITH 1186 00:51:51,022 --> 00:51:53,391 ME IN THE BEGINNING. 1187 00:51:53,391 --> 00:51:54,192 (INDISCERNIBLE) DECIDED TO 1188 00:51:54,192 --> 00:51:56,428 RETIRE THIS YEAR AND I'M LARGELY 1189 00:51:56,428 --> 00:51:59,865 HEART BROKEN OVER THAT 1190 00:51:59,865 --> 00:52:01,800 REVELATION AND AUGUSTA FRANKO 1191 00:52:01,800 --> 00:52:03,135 LOST HIS LIFE TO LYMPHOMA MANY 1192 00:52:03,135 --> 00:52:04,402 THE EARLY 2000s BUT THE WORK 1193 00:52:04,402 --> 00:52:08,573 IN GENETICS OF ETBF STAINS TODAY 1194 00:52:08,573 --> 00:52:10,175 -- SPANS TODAY. THERE ARE MANY 1195 00:52:10,175 --> 00:52:13,478 OTHER PEOPLE CURRENT LAB AND 1196 00:52:13,478 --> 00:52:15,113 PRIOR LABS THAT CONTRIBUTED TO 1197 00:52:15,113 --> 00:52:17,482 THIS PROJECT AS WELL AS NUMEROUS 1198 00:52:17,482 --> 00:52:21,219 PEOPLE WHO COLLABORATED WITH US 1199 00:52:21,219 --> 00:52:23,188 OVER TIME. I OWE A DEEP 1200 00:52:23,188 --> 00:52:26,858 GRATITUDE TO THE PEOPLE WHO 1201 00:52:26,858 --> 00:52:28,293 INSTITUTIONS THAT FUNDED US AND 1202 00:52:28,293 --> 00:52:29,427 THAT IT IS THE NIH IS THE 1203 00:52:29,427 --> 00:52:34,065 FOUNDATION OF THAT. IT IS MOSTLY 1204 00:52:34,065 --> 00:52:37,202 NIDDK AND CI I DID GET ONE AND 1205 00:52:37,202 --> 00:52:39,771 NAIA GRANT AT ONE TIM TIME. LAST 1206 00:52:39,771 --> 00:52:41,373 NOT LEAST, ALL THE GENEROUS 1207 00:52:41,373 --> 00:52:42,440 PATIENTS WHO HAVE WORKED WITH 1208 00:52:42,440 --> 00:52:49,181 US. ONE DAY I CAME BACK INDEED 1209 00:52:49,181 --> 00:52:50,782 MY DOOR HAD BEEN ADORING AND IT 1210 00:52:50,782 --> 00:52:54,386 IS NOW BEEN THERE AT LEAST A 1211 00:52:54,386 --> 00:52:57,189 DECADE AND OF COURSE WORK 1212 00:52:57,189 --> 00:52:57,923 REMAINS ABSOLUTELY TRUE. THANK 1213 00:52:57,923 --> 00:52:59,691 YOU VERY MUCH FOR LISTENING. BE 1214 00:52:59,691 --> 00:53:02,961 HAPPY TO ANSWER ANY QUESTIONS. 1215 00:53:02,961 --> 00:53:05,030 [APPLAUSE] 1216 00:53:05,030 --> 00:53:09,434 >> ANYONE FREE TO QUESTIONS, 1217 00:53:09,434 --> 00:53:10,602 PLEASE YOU CAN KICK US OFF. 1218 00:53:10,602 --> 00:53:17,042 >> THANK YOU SO MUCH. 1219 00:53:17,042 --> 00:53:19,311 >> WAITING FOR IT TO TURN ON. 1220 00:53:19,311 --> 00:53:23,748 >> MY QUESTION RELATES TO THE 1221 00:53:23,748 --> 00:53:26,551 END STIMULUS. . DO YOU THINK THE 1222 00:53:26,551 --> 00:53:30,922 BUGS AND TOXINS YOU DESCRIBED 1223 00:53:30,922 --> 00:53:33,859 ARE DIRECTLY STIMULATING THE 1224 00:53:33,859 --> 00:53:36,561 COLON CELLS TO TRANSFORM INTO 1225 00:53:36,561 --> 00:53:39,898 MALIGNANCY? OR AS YOU SUGGESTED 1226 00:53:39,898 --> 00:53:42,167 SEVERAL TIMES, ARE THEY CREATING 1227 00:53:42,167 --> 00:53:45,270 THIS MASSIVE INFLAMMATORY 1228 00:53:45,270 --> 00:53:47,706 REACTION WHOSE REACTIVE OXYGEN 1229 00:53:47,706 --> 00:53:48,473 SPECIES AND OTHER PRODUCTS ARE 1230 00:53:48,473 --> 00:53:50,342 INDUCING THE TRANSFORMATION? 1231 00:53:50,342 --> 00:53:53,812 >> I THINK THIS IS A BIG DEBATE 1232 00:53:53,812 --> 00:53:55,513 IN THE FIELD. SO THE ONE 1233 00:53:55,513 --> 00:53:59,484 ORGANISM ON THAT LIST BEING 1234 00:53:59,484 --> 00:54:02,053 CONSIDERED AS POSSIBLY TARGETING 1235 00:54:02,053 --> 00:54:04,623 THE DNA, THAT TRANSFORMING THE 1236 00:54:04,623 --> 00:54:08,627 CELLS IS THE COLON MADE BY 1237 00:54:08,627 --> 00:54:11,263 POLYSYNTHASE E. COLI. I HOWEVER, 1238 00:54:11,263 --> 00:54:13,798 DON'T THINK THE EVIDENCE HOLDS 1239 00:54:13,798 --> 00:54:15,834 UP FOR THAT CONCLUSION AS YET. I 1240 00:54:15,834 --> 00:54:17,969 THINK THESE ORGANISMS ARE 1241 00:54:17,969 --> 00:54:20,572 PROMOTIONAL. THEN IN FACT I 1242 00:54:20,572 --> 00:54:21,740 DON'T KNOW WHAT CAUSES THE 1243 00:54:21,740 --> 00:54:24,542 MUTATION OF APC THAT'S 1244 00:54:24,542 --> 00:54:26,478 FOUNDATIONAL TO COLON CANCER BUT 1245 00:54:26,478 --> 00:54:28,914 I REALLY THINK THE BUGS ARE 1246 00:54:28,914 --> 00:54:30,348 COMING IN CREATING AS YOU JUST 1247 00:54:30,348 --> 00:54:34,853 SAID. THE CONTEXT FOR 1248 00:54:34,853 --> 00:54:37,289 DEVELOPMENT OF CANCER. 1249 00:54:37,289 --> 00:54:39,024 >> IT WOULD BE EASY WITH USING 1250 00:54:39,024 --> 00:54:41,826 THE TOXINS TO SEE WHAT AFFECT 1251 00:54:41,826 --> 00:54:46,998 THEY HAVE FOR EXAMPLE ON 1252 00:54:46,998 --> 00:54:47,999 PHAGOSITIC CELLS AND OTHER CELLS 1253 00:54:47,999 --> 00:54:50,702 TO GENERATE A HUGE GENERATION OF 1254 00:54:50,702 --> 00:54:52,370 REACTIVE OXYGEN SPECIES. 1255 00:54:52,370 --> 00:54:58,176 >> WE DID THAT ONCE WITH JERRY 1256 00:54:58,176 --> 00:54:58,944 DONOWITS YOU PROBABLY KNOW 1257 00:54:58,944 --> 00:55:01,947 TRYING TO LOOK AT BFT EFFECT ON 1258 00:55:01,947 --> 00:55:06,851 POLYS. THERE WAS NO EFFECT THAT 1259 00:55:06,851 --> 00:55:10,255 WE COULD DETECT. IF TESTIFY 1260 00:55:10,255 --> 00:55:12,457 CLAUDIN 4 IS RECEPTOR FOR BFT I 1261 00:55:12,457 --> 00:55:14,025 DON'T THINK IT IS ON MOST OF 1262 00:55:14,025 --> 00:55:16,928 THOSE CELLS ANYWAY. SO AT LEAST 1263 00:55:16,928 --> 00:55:18,396 WITH THAT TOXIN, WE HAVEN'T 1264 00:55:18,396 --> 00:55:21,866 STUDIED THE OTHERS, I DON'T 1265 00:55:21,866 --> 00:55:23,268 THINK IT IS DIRECT ON THE IMMUNE 1266 00:55:23,268 --> 00:55:26,171 SYSTEM, I THINK IT IS A CASCADE 1267 00:55:26,171 --> 00:55:29,541 FROM THE LUMIN TO SUB MUCOSA 1268 00:55:29,541 --> 00:55:32,377 BACK TO EPITHELIAL CELLS, AT 1269 00:55:32,377 --> 00:55:35,880 LEAST WITH ETBF, IT IS THAT 1270 00:55:35,880 --> 00:55:38,583 FEEDBACK OF IL 17 ON THE 1271 00:55:38,583 --> 00:55:40,352 EPITHELIAL SIDE THAT SEEMS TO 1272 00:55:40,352 --> 00:55:43,922 TRIGGER THE EPIPEELIAL CELL TO 1273 00:55:43,922 --> 00:55:44,356 CHANGE ITS FUNCTION. 1274 00:55:44,356 --> 00:55:45,924 >> THANK YOU. 1275 00:55:45,924 --> 00:55:48,593 >> COUPLE OF QUESTIONS ONLINE. 1276 00:55:48,593 --> 00:55:52,030 SO DID THE ANTIBIOTIC DATA HAVE 1277 00:55:52,030 --> 00:55:53,965 SUFFICIENT DETAIL TO SEE IF SOME 1278 00:55:53,965 --> 00:55:55,367 ANTIBIOTICS ARE ASSOCIATED WITH 1279 00:55:55,367 --> 00:55:57,402 HIGHER COLON RISK COMPARED TO 1280 00:55:57,402 --> 00:55:58,837 OTHER ANTIBIOTICS? 1281 00:55:58,837 --> 00:56:02,073 >> I DIDN'T SAY THIS BUT WHEN WE 1282 00:56:02,073 --> 00:56:03,875 DRILLED DOWN IN TO THAT 1283 00:56:03,875 --> 00:56:06,344 QUESTION, IT TURNED OUT THAT THE 1284 00:56:06,344 --> 00:56:09,014 ASSOCIATED ANTIBIOTIC WAS 1285 00:56:09,014 --> 00:56:13,451 PENICILLIN OF ALL THINGS. I MUST 1286 00:56:13,451 --> 00:56:16,121 ADMIT I WAS SURPRISED BUT 1287 00:56:16,121 --> 00:56:19,657 PENICILLIN HAS A VARIETY OF 1288 00:56:19,657 --> 00:56:22,894 ACTIVITIES AGAINST AN AROBES AND 1289 00:56:22,894 --> 00:56:24,029 PARTICULARLY WHEN -- EVERY TIME 1290 00:56:24,029 --> 00:56:26,064 YOU TAKE AN ORAL ANTIBIOTIC IT 1291 00:56:26,064 --> 00:56:28,400 IS NOT COMPLETELY ABSORBED. 1292 00:56:28,400 --> 00:56:31,369 EVERY TIME IT HITS THE COLON AND 1293 00:56:31,369 --> 00:56:33,605 MODIFIES THE MICROBIOME. IT IS 1294 00:56:33,605 --> 00:56:36,674 AN IMPOSSIBLE QUESTION TO STUDY 1295 00:56:36,674 --> 00:56:40,311 BUT I THINK THOSE SERIAL SHIFTS 1296 00:56:40,311 --> 00:56:41,646 SOMEHOW CHANGE THE BIOLOGY OF 1297 00:56:41,646 --> 00:56:43,481 THE MUCOUS LAYER THERE, MAKE IT 1298 00:56:43,481 --> 00:56:46,351 MORE AMENABLE TO BIOFILM 1299 00:56:46,351 --> 00:56:50,088 INVASION IN SOME OF THE AFFECTS 1300 00:56:50,088 --> 00:56:53,725 THAT WE HAVE SEEN. THAT RESULT 1301 00:56:53,725 --> 00:56:56,161 GOT FURTHER REINFORCED WHEN WE 1302 00:56:56,161 --> 00:56:58,897 LOOKED AT PEOPLE THAT ONLY HAD 1303 00:56:58,897 --> 00:57:01,066 EVER TAKEN PENICILLIN VERSUS ANY 1304 00:57:01,066 --> 00:57:02,500 OTHER ANTIBIOTIC AND ACTUALLY 1305 00:57:02,500 --> 00:57:04,803 THE RESULTS HELD UP EVEN UNDER 1306 00:57:04,803 --> 00:57:07,405 THOSE CIRCUMSTANCES. 1307 00:57:07,405 --> 00:57:09,641 >> ANOTHER QUESTION, MAY HAVE 1308 00:57:09,641 --> 00:57:11,342 ANTICIPATED THIS ANSWER, WERE 1309 00:57:11,342 --> 00:57:14,779 THERE FUNGI IN THE BIOFILMS? 1310 00:57:14,779 --> 00:57:17,348 >> I HAPPEN TO LIKE BACTERIA, SO 1311 00:57:17,348 --> 00:57:22,353 WE REALLY FOCUSED BACTERIA, WE 1312 00:57:22,353 --> 00:57:24,355 HAVEN'T DEALT, THERE ARE FEW 1313 00:57:24,355 --> 00:57:26,191 PAPERS OUT THERE NOW, I THINK WE 1314 00:57:26,191 --> 00:57:29,294 HAVE A LOT TO LEARN ABOUT 1315 00:57:29,294 --> 00:57:31,896 VIRUSES PARASITES AND FUNGI HOW 1316 00:57:31,896 --> 00:57:36,734 THEY MAY INFLUENCE COLON CANCER. 1317 00:57:36,734 --> 00:57:42,273 >> THAT IS IT ONLINE. DR. 1318 00:57:42,273 --> 00:57:42,507 HADIGAN? 1319 00:57:42,507 --> 00:57:44,709 >> I JUST WANT TO THANK DR. 1320 00:57:44,709 --> 00:57:47,745 SEARS AGAIN FOR A FANTASTIC 1321 00:57:47,745 --> 00:57:49,347 LECTURE. THANKS EVERYBODY FOR 1322 00:57:49,347 --> 00:57:50,281 ATTENDING AND LET THOSE 1323 00:57:50,281 --> 00:57:51,516 INDIVIDUAL WHOSE ARE PRESENT 1324 00:57:51,516 --> 00:57:54,219 HERE KNOW THERE IS A RECEPTION 1325 00:57:54,219 --> 00:57:55,453 OUTSIDE LIPSETT, HOPE YOU WILL 1326 00:57:55,453 --> 00:57:56,921 BE ABLE TO JOIN US AND MAYBE 1327 00:57:56,921 --> 00:57:58,189 HAVE AN OPPORTUNITY TO MEET DR. 1328 00:57:58,189 --> 00:58:00,125 SEARS. THANK YOU, EVERYBODY. 1329 00:58:00,125 --> 00:58:10,301 [APPLAUSE]