1 00:00:05,960 --> 00:00:07,760 >> WELCOME EVERYBODY, I'M 2 00:00:07,760 --> 00:00:09,360 NINA SHORE AND I'M THE DEPUTY 3 00:00:09,360 --> 00:00:10,360 DIRECTOR AND ACTING 4 00:00:10,360 --> 00:00:12,680 SCIENTIFIC DIRECTOR OF NINDS AND 5 00:00:12,680 --> 00:00:14,280 IT IS TRULY MY HONOR AND 6 00:00:14,280 --> 00:00:18,280 PLEASURE TO WELCOME YOU TO THIS 7 00:00:18,280 --> 00:00:19,160 WEDNESDAY AFTERNOON LECTURE. 8 00:00:19,160 --> 00:00:21,240 THOSE OF YOU WHO ARE ON 9 00:00:21,240 --> 00:00:25,040 VIDEOCAST, MAY NOT REALIZE THIS, 10 00:00:25,040 --> 00:00:29,040 BUT THIS IS THE FIRST MAJOR NIH 11 00:00:29,040 --> 00:00:32,840 EVENT THAT IS OCCURRING IN 12 00:00:32,840 --> 00:00:36,240 PERSON HERE AT 13 00:00:36,240 --> 00:00:37,080 LIPSETT AMPHITHEATRE. 14 00:00:37,080 --> 00:00:40,640 WE HAVE A VERY SMALL AND VERY 15 00:00:40,640 --> 00:00:41,640 SOCIALLY DISTANCED AUDIENCE AND 16 00:00:41,640 --> 00:00:44,720 IT IS A REALLY EXCITING MOMENT 17 00:00:44,720 --> 00:00:47,680 AFTER 2 YEARS OF EVERYTHING 18 00:00:47,680 --> 00:00:48,960 BEING VIDEOCAST. 19 00:00:48,960 --> 00:00:49,840 SO WELCOME WHETHER YOU ARE HERE 20 00:00:49,840 --> 00:00:55,080 WITH US IN THE UDIANCY OR ON 21 00:00:55,080 --> 00:00:55,720 VIDEOCAST TODAY. 22 00:00:55,720 --> 00:00:57,400 I AM VERY EXCITED AND YOU WILL 23 00:00:57,400 --> 00:00:59,640 HEAR IN A MOMENT WHY IT'S ALSO A 24 00:00:59,640 --> 00:01:02,160 PERSONAL PLEASURE FOR ME TO 25 00:01:02,160 --> 00:01:07,240 INTRODUCE TODAY'S SPEAKER 26 00:01:07,240 --> 00:01:11,040 DR. ANNA HUTTENLOCHER, 27 00:01:11,040 --> 00:01:11,840 DR. HUTTENLOCHER PUT ON THE MAP 28 00:01:11,840 --> 00:01:13,680 THE ROLE OF CELL MIGRATION IN 29 00:01:13,680 --> 00:01:16,800 TISSUE REPAIR AND IN THE REPAIR 30 00:01:16,800 --> 00:01:20,560 OF INFLAMMATION 31 00:01:20,560 --> 00:01:22,360 DR. ANNA HUTTENLOCHER IS A 32 00:01:22,360 --> 00:01:23,600 PROFESSOR OF PEDIATRICS AND 33 00:01:23,600 --> 00:01:24,840 MEDICAL MICROBIOLOGY AND 34 00:01:24,840 --> 00:01:28,200 IMMUNOLOGY AT THE UNIVERSITY OF 35 00:01:28,200 --> 00:01:28,920 WISCONSIN MADISON. 36 00:01:28,920 --> 00:01:31,360 AS I SAID, HER RESEARCH REALLY 37 00:01:31,360 --> 00:01:34,240 HAS FOCUSED FOR MANY YEARS ON 38 00:01:34,240 --> 00:01:37,760 THE MOLECULAR MECHANISMS AND THE 39 00:01:37,760 --> 00:01:40,360 MORE MORPHOLOGY OF CELL 40 00:01:40,360 --> 00:01:44,120 MIGRATION IN RELATION TO THE 41 00:01:44,120 --> 00:01:48,000 REPAIR OF INFLAMED AND INJURED 42 00:01:48,000 --> 00:01:48,720 TISSUES. 43 00:01:48,720 --> 00:01:50,200 SHE RECEIVED HER UNDERGRADUATE 44 00:01:50,200 --> 00:01:53,760 DEGREE IN BIOLOGY FROM OBERLAND 45 00:01:53,760 --> 00:01:56,160 IN COLLEGE AND HER MEDICAL 46 00:01:56,160 --> 00:01:59,120 DEGREE FROM HARVARD MEDICAL 47 00:01:59,120 --> 00:01:59,920 SCHOOL. 48 00:01:59,920 --> 00:02:02,760 AS A YALEER AND OTHER THAN THAT 49 00:02:02,760 --> 00:02:04,520 SHE HAD AN UNBLEMMISHED CAREER. 50 00:02:04,520 --> 00:02:07,200 HE WENT ON TO DO HER RESIDENCY 51 00:02:07,200 --> 00:02:09,040 AT PEDIATRICS AT UNIVERSITY OF 52 00:02:09,040 --> 00:02:11,360 CALIFORNIA SAN FRANCISCO AND 53 00:02:11,360 --> 00:02:13,520 POST DOCTORAL FELLOWSHIPS BOTH 54 00:02:13,520 --> 00:02:19,960 AT UCSF AND AT UNIVERSITY OF 55 00:02:19,960 --> 00:02:20,560 ILLINOIS URBANA CHAMPAGNE. 56 00:02:20,560 --> 00:02:22,880 SHE MOVED TO THE UNIVERSITY OF 57 00:02:22,880 --> 00:02:25,720 WISCONSIN MEDICAL SCHOOL AS AN 58 00:02:25,720 --> 00:02:27,160 ASSISTANT PROFESSOR IN 1999 AND 59 00:02:27,160 --> 00:02:29,000 HAS BEEN THERE EVER SINCE. 60 00:02:29,000 --> 00:02:32,560 AND SHE NOW HAS AN ENDOWED 61 00:02:32,560 --> 00:02:38,560 PROFESSORSHIP AND RUNS A VERY 62 00:02:38,560 --> 00:02:39,000 ILLUSTRIOUS LABRATORY 63 00:02:39,000 --> 00:02:39,600 ENTERPRISE. 64 00:02:39,600 --> 00:02:42,440 SHE HAS WON MANY, MANY AWARDS 65 00:02:42,440 --> 00:02:44,520 AND HAS MANY ACCOLADES INCLUDING 66 00:02:44,520 --> 00:02:47,240 BEING A MEMBER OF THE NATIONAL 67 00:02:47,240 --> 00:02:48,680 ACADEMY OF MEDICINE AND THE 68 00:02:48,680 --> 00:02:51,480 ASSOCIATION OF AMERICAN 69 00:02:51,480 --> 00:02:52,760 PHYSICIANS. 70 00:02:52,760 --> 00:02:59,760 SHE IS A RECIPIENT OF THE BURROW 71 00:02:59,760 --> 00:03:00,760 SIZE AMERICAN MEDICAL SCIENTIST 72 00:03:00,760 --> 00:03:01,920 AWARD AND SHE HAS BEEN WITHIN 73 00:03:01,920 --> 00:03:03,600 THE LAST YEAR, I BELIEVE, A 74 00:03:03,600 --> 00:03:07,560 MEMBER OF THE BSC OF THE NIAMS. 75 00:03:07,560 --> 00:03:12,480 SO SHE IS NO STRANGER TO NIH AND 76 00:03:12,480 --> 00:03:13,800 IT'S INTRAMURAL PROGRAM AND JUST 77 00:03:13,800 --> 00:03:16,760 ON A PERSONAL NOTE, I FIRST GOT 78 00:03:16,760 --> 00:03:18,320 TO KNOW ANNA IN THE CONTEXT OF 79 00:03:18,320 --> 00:03:21,040 BOTH OF US BEING ON THE STEERING 80 00:03:21,040 --> 00:03:24,760 COMMITTEE OF SOMETHING CALLED 81 00:03:24,760 --> 00:03:25,280 THE PEDIATRIC SCIENTIST 82 00:03:25,280 --> 00:03:26,560 DEVELOPMENT PROGRAM, A PROGRAM 83 00:03:26,560 --> 00:03:30,120 THAT'S FUNDED FOR MANY YEARS BY 84 00:03:30,120 --> 00:03:31,680 NICHD. 85 00:03:31,680 --> 00:03:32,640 AND BECAUSE OF THAT ASSOCIATION, 86 00:03:32,640 --> 00:03:37,120 I KNOW HER TO REALLY BE 87 00:03:37,120 --> 00:03:38,760 PASSIONATE ABOUT FUELING, 88 00:03:38,760 --> 00:03:41,720 EDUCATING, MENTORING AND 89 00:03:41,720 --> 00:03:43,760 LAUNCHING THE NEXT GENERATION OF 90 00:03:43,760 --> 00:03:45,080 SCIENTISTS AND PHYSICIANS, AND 91 00:03:45,080 --> 00:03:46,120 PHYSICIAN SCIENTISTS, AND SO 92 00:03:46,120 --> 00:03:48,080 IT'S WITH GREAT PLEASURE THAT I 93 00:03:48,080 --> 00:03:57,920 TURN THE PODIUM OVER TO 94 00:03:57,920 --> 00:03:58,320 DR. HUTTENLOCHER. 95 00:03:58,320 --> 00:03:59,600 >> THANK YOU VERY MUCH. 96 00:03:59,600 --> 00:04:00,960 IT'S TRULY AN HONOR TO BE 97 00:04:00,960 --> 00:04:02,520 INVITED TO DO THIS AND IT'S 98 00:04:02,520 --> 00:04:06,720 TRULY AN HONOR TO BE GIVING THE 99 00:04:06,720 --> 00:04:07,960 FIRST HYBRID IN-PERSON TALK AT 100 00:04:07,960 --> 00:04:11,040 NIH IN OVER 2 YEARS AND TO BE 101 00:04:11,040 --> 00:04:12,520 HONEST IT'S MY FIRST TRIP TO 102 00:04:12,520 --> 00:04:16,760 SPEAK IN PERSON, SO THANK YOU 103 00:04:16,760 --> 00:04:17,720 VERY MUCH. 104 00:04:17,720 --> 00:04:18,960 THIS IS FANTASTIC, SO I WILL BE 105 00:04:18,960 --> 00:04:20,520 TALKING TO YOU TODAY ABOUT OUR 106 00:04:20,520 --> 00:04:23,240 LONG-TERM INTERESTS IN 107 00:04:23,240 --> 00:04:23,920 UNDERSTANDING LEUCOCYTE MOTILITY 108 00:04:23,920 --> 00:04:28,240 IN TISSUES AND WHAT THIS MEANS 109 00:04:28,240 --> 00:04:30,040 FOR INFLAMMATION, IN PARTICULAR, 110 00:04:30,040 --> 00:04:32,160 WORK FOCUSED ON THE RESOLUTION 111 00:04:32,160 --> 00:04:32,960 OF INFLAMMATION AND THE CELL 112 00:04:32,960 --> 00:04:35,280 TYPE THAT I WILL BE FOCUSING ON 113 00:04:35,280 --> 00:04:38,560 TO A LARGE EXTENT IS THE 114 00:04:38,560 --> 00:04:39,080 NUTRIFILL. 115 00:04:39,080 --> 00:04:41,680 SO NUTRIFILLS ARE PRIMARY CELLS 116 00:04:41,680 --> 00:04:43,920 OF THE INNATE IMMUNE RESPONSE 117 00:04:43,920 --> 00:04:44,840 AND THEY'RE THE FIRST RESPONDERS 118 00:04:44,840 --> 00:04:46,440 TO TISSUE DAMAGE AND IN THIS 119 00:04:46,440 --> 00:04:49,120 VERY CLASSIC MOVIE OF A NEWT 120 00:04:49,120 --> 00:04:50,880 FILL RESPONDING TO A BACTERIUM, 121 00:04:50,880 --> 00:04:53,160 CAN YOU SEE THEIR ABILITY TO 122 00:04:53,160 --> 00:05:11,720 MIGRATE AND GRATE AND CHANGE DIRECTION 123 00:05:11,720 --> 00:05:12,920 CONVICTLY--CONTRIBUTES TO 124 00:05:12,920 --> 00:05:14,280 DEVELOPMENT OF AUTOIMMUNITY SO 125 00:05:14,280 --> 00:05:16,160 REALLY WE'VE BEEN INTERESTED IN 126 00:05:16,160 --> 00:05:18,480 THIS QUESTION OF HOW NEUTROPHILS 127 00:05:18,480 --> 00:05:20,560 GET SITES OF TISSUE DAMAGE 128 00:05:20,560 --> 00:05:22,440 VERSUS INFECTION, YOU WANT THEM 129 00:05:22,440 --> 00:05:23,960 AT INFECTED TISSUES BUT YOU 130 00:05:23,960 --> 00:05:26,720 DON'T WANT THEM TO STAY AT SITES 131 00:05:26,720 --> 00:05:27,760 OF TISSUE DAMAGE. 132 00:05:27,760 --> 00:05:29,760 SO AT A REVIEW ARTICLE WE WROTE 133 00:05:29,760 --> 00:05:31,760 A NUMBER OF YEARS AGO, THERE ARE 134 00:05:31,760 --> 00:05:32,960 FACTORS INVOLVED IN THE 135 00:05:32,960 --> 00:05:33,760 RECRUITMENT AND MANY RESEARCH 136 00:05:33,760 --> 00:05:37,720 GROUP VS WORKED ON THIS IN TERMS 137 00:05:37,720 --> 00:05:41,400 OF DAMPS, WOUNDS THAT MEDIATE 138 00:05:41,400 --> 00:05:42,600 NEUTROPHIL IMPROVEMENT, CHEMO 139 00:05:42,600 --> 00:05:45,720 KINE RECEPTOR SIGNALING AND THEN 140 00:05:45,720 --> 00:05:46,920 THIS WHOLE INTERESTING 141 00:05:46,920 --> 00:05:48,440 PHENOMENON OF NEUTROPHILS 142 00:05:48,440 --> 00:05:51,240 RELEASING ATTRACT ANTS LIKE LTB4 143 00:05:51,240 --> 00:05:53,720 WHICH LEAD TO SWARMING AND LEAD 144 00:05:53,720 --> 00:05:54,440 TO INCREASED FORMATION AND 145 00:05:54,440 --> 00:05:55,600 TISSUES AND THIS IS WHAT HAPPENS 146 00:05:55,600 --> 00:05:59,520 IN THE KIND OF STERILE TISSUE 147 00:05:59,520 --> 00:05:59,760 DAMAGE. 148 00:05:59,760 --> 00:06:02,680 WITH INFECTION YOU ADD 149 00:06:02,680 --> 00:06:04,600 ADDITIONAL COMPONENTS AND P A 150 00:06:04,600 --> 00:06:05,160 MPs AND MICROORGANISM THAT 151 00:06:05,160 --> 00:06:07,120 SAYS LEAD TO THE ACTIVATION OF 152 00:06:07,120 --> 00:06:08,160 DIFFERENT SIGNALING PATHWAYS AND 153 00:06:08,160 --> 00:06:10,960 WE WERE EXPWED IN HAD 154 00:06:10,960 --> 00:06:11,960 DISTINGUISHED NEUTROPHIL 155 00:06:11,960 --> 00:06:14,320 RESPONSE TO TISSUE DAMAGE AND 156 00:06:14,320 --> 00:06:17,200 MICROBIAL SIGNALS AND IN 157 00:06:17,200 --> 00:06:19,920 PARTICULAR WHAT SIGNALS PROMOTE 158 00:06:19,920 --> 00:06:20,440 RESOLUTION OF NEUTROPHIL 159 00:06:20,440 --> 00:06:22,040 INFLAMMATION AND FOR MUCH OF 160 00:06:22,040 --> 00:06:24,480 THIS WORK, WE USE ZEBRA FISH AS 161 00:06:24,480 --> 00:06:26,960 A MODEL SYSTEM, SO WHY DO WE USE 162 00:06:26,960 --> 00:06:27,720 ZEBRA FISH? 163 00:06:27,720 --> 00:06:31,240 BECAUSE YOU CAN WATCH MOTILITY 164 00:06:31,240 --> 00:06:35,080 WITH HIGH ABILITY INVIVO BECAUSE 165 00:06:35,080 --> 00:06:36,400 THE ZEBRA FISH ARE TRANSPARENT. 166 00:06:36,400 --> 00:06:40,160 AND YOU CAN SEE HERE AT 72 HOURS 167 00:06:40,160 --> 00:06:41,560 POST FERTILIZATION, CAN YOU SEE 168 00:06:41,560 --> 00:06:43,480 THAT NUTRIFILL VS 2 POPULATIONS. 169 00:06:43,480 --> 00:06:47,880 SO THESE RAPIDLY MODEL CITIZEN 170 00:06:47,880 --> 00:06:49,200 TILE AMOEBOID NEUTROPHILS IN THE 171 00:06:49,200 --> 00:06:51,720 HEAD REGION AND THEN MUCH MORE 172 00:06:51,720 --> 00:06:54,760 STATIONARY NUTRIFILLS WITHIN THE 173 00:06:54,760 --> 00:06:56,560 TISSUE AND TISSUE DAMAGE THEY'RE 174 00:06:56,560 --> 00:06:57,720 RAPIDLY MOBILIZED AND LEAD THE 175 00:06:57,720 --> 00:07:01,040 CHT AND THIS IS VERY DIFFERENT 176 00:07:01,040 --> 00:07:02,560 FROM MACROPHAGES AND THEY 177 00:07:02,560 --> 00:07:08,240 EXHIBIT THIS SURVEILLANCE 178 00:07:08,240 --> 00:07:08,520 MIGRATION. 179 00:07:08,520 --> 00:07:10,120 THEIR MYRRHIGRATION IS PROTEASE 180 00:07:10,120 --> 00:07:11,120 INDEPENDENT AND THEY NIGREAT 181 00:07:11,120 --> 00:07:13,200 MORE IN A MESSENTERIEN CHIMAL 182 00:07:13,200 --> 00:07:14,440 TYPE MODE OF MIGRATION AND SO 183 00:07:14,440 --> 00:07:15,800 WE'RE INTERESTED IN 184 00:07:15,800 --> 00:07:18,240 UNDERSTANDING THESE DIFFERENT 185 00:07:18,240 --> 00:07:20,560 MECHANISMS OF MIGRATION AND IN 186 00:07:20,560 --> 00:07:22,160 ADDITION WITH NEUTROPHILS WHAT 187 00:07:22,160 --> 00:07:23,560 MEDIATES THE RECRUITMENT TO 188 00:07:23,560 --> 00:07:24,400 TISSUE DAMAGE. 189 00:07:24,400 --> 00:07:25,920 SO THIS KIND OF DEMONSTRATES THE 190 00:07:25,920 --> 00:07:28,960 POWER OF THE FISH SYSTEM SO WE 191 00:07:28,960 --> 00:07:31,240 CAN BOTH CUT OFF THE TAIL AND 192 00:07:31,240 --> 00:07:33,040 INJECT BACTERIA INTO THE EAR 193 00:07:33,040 --> 00:07:34,520 REGION OF THE ZEBRA FISH AND YOU 194 00:07:34,520 --> 00:07:35,720 CAN SEE NEUTROPHILS ARE 195 00:07:35,720 --> 00:07:37,720 RECRUITED BOTH TO THE INFECTION 196 00:07:37,720 --> 00:07:39,720 AND TO THE SITE OF STERILE 197 00:07:39,720 --> 00:07:44,320 INJURY AND IF YOU ADD AN 198 00:07:44,320 --> 00:07:45,920 INHIBITOR TO NADPHOX DACE IN 199 00:07:45,920 --> 00:07:47,360 THIS CASE, DPI, YOU BROCK 200 00:07:47,360 --> 00:07:49,120 RECRUITMENT TO THE STERILE 201 00:07:49,120 --> 00:07:50,200 INJURY SITE BUT YOU DO NOT BLOCK 202 00:07:50,200 --> 00:07:52,240 IT TO THE SITE OF INFECTION AND 203 00:07:52,240 --> 00:07:55,560 IN FACT, YOU SEE MORE NEUTROPHIL 204 00:07:55,560 --> 00:07:56,240 RECRUITMENT. 205 00:07:56,240 --> 00:07:58,240 SO THIS DEMONSTRATES NICELY THAT 206 00:07:58,240 --> 00:07:59,200 DIFFERENT SIGNALS MEDIATE 207 00:07:59,200 --> 00:08:00,280 NEUTROPHIL RECRUITMENT TO WOUNDS 208 00:08:00,280 --> 00:08:00,720 AND INFECTION. 209 00:08:00,720 --> 00:08:02,240 SO WE WERE INTERESTED IN TRYING 210 00:08:02,240 --> 00:08:05,160 TO FIGURE OUT SOME OF THESE 211 00:08:05,160 --> 00:08:05,600 DIFFERENTIAL SIGNALS. 212 00:08:05,600 --> 00:08:06,840 SO I WOULD LIKE TO FIRST 213 00:08:06,840 --> 00:08:08,000 HIGHLIGHT THOUGH THAT IN 214 00:08:08,000 --> 00:08:09,560 ADDITION TO BEING A GREAT SYSTEM 215 00:08:09,560 --> 00:08:15,040 TO LOOK AT MOTILITY, THE INNATE 216 00:08:15,040 --> 00:08:16,840 SYSTEM IS HIGHLY CONSERVED SO 217 00:08:16,840 --> 00:08:19,200 CAN YOU MODEL HUMAN DISEASES IN 218 00:08:19,200 --> 00:08:19,520 ZEBRA FISH. 219 00:08:19,520 --> 00:08:21,040 AN EXAMPLE FOR MANY YEARS AGO IS 220 00:08:21,040 --> 00:08:22,880 AN INFANT BOY WE SAW AT THE 221 00:08:22,880 --> 00:08:24,840 UNIVERSITY OF WISCONSIN WHO 222 00:08:24,840 --> 00:08:26,320 PRESENTED WITH RECURRENT 223 00:08:26,320 --> 00:08:29,160 BACTERIAL INFECTIONS AND WE USED 224 00:08:29,160 --> 00:08:30,360 MICROFLUIDICS IN COLLABORATION 225 00:08:30,360 --> 00:08:33,640 WITH THE BB LAB AND YOU CAN SEE 226 00:08:33,640 --> 00:08:35,840 NICE CHEMO TAXIS OF CONTROL 227 00:08:35,840 --> 00:08:37,640 NEUTROPHILS AND SIGNIFICANTLY 228 00:08:37,640 --> 00:08:39,120 IMPAIRED MOTILITY OF PATIENT 229 00:08:39,120 --> 00:08:39,520 NEUTROPHILS. 230 00:08:39,520 --> 00:08:40,440 AND THIS PATIENT WAS FOUND TO 231 00:08:40,440 --> 00:08:43,160 HAVE A FORNL OF LEUCOCYTE 232 00:08:43,160 --> 00:08:43,560 ADHESION DEFICIENCY. 233 00:08:43,560 --> 00:08:49,160 IN THIS CASE, IT WAS A DOMINANT 234 00:08:49,160 --> 00:08:52,440 INHIBITORY MUTATION IN THE SMALL 235 00:08:52,440 --> 00:08:55,560 DTPACE RAB 2, NOW RAC2 AND 236 00:08:55,560 --> 00:08:56,560 HIGHLY CONSERVED IN ZEBRA FISH 237 00:08:56,560 --> 00:08:59,680 SO WE CAN TAKE ADVANTAGE OF THIS 238 00:08:59,680 --> 00:09:02,480 CONSERVATION TO INTRODUCE THE 239 00:09:02,480 --> 00:09:03,520 MUTATION SPECIFICALLY INTO 240 00:09:03,520 --> 00:09:03,800 NEUTROPHILS. 241 00:09:03,800 --> 00:09:07,200 SO WHAT IS KNOWN ABOUT RAC2 242 00:09:07,200 --> 00:09:08,880 SIGNALING AND THEIR OWN 243 00:09:08,880 --> 00:09:09,160 MOTILITY. 244 00:09:09,160 --> 00:09:11,160 SO DOWN STREAM OF CHEMO KINE 245 00:09:11,160 --> 00:09:14,760 SIGNALING YOU GET ACTIVATION OF 246 00:09:14,760 --> 00:09:15,760 G-PROTEIN COUPLED RECEPTORS, 247 00:09:15,760 --> 00:09:18,320 GAPS THAT LEAD TO RAC, GTPACE 248 00:09:18,320 --> 00:09:19,720 ACTIVATION OF THE LEADING EDGE 249 00:09:19,720 --> 00:09:21,520 WHICH IS A CRITICAL REGULATOR OF 250 00:09:21,520 --> 00:09:23,760 DYNAMIC ACTIN AT THE LEADING 251 00:09:23,760 --> 00:09:24,640 EDGE. 252 00:09:24,640 --> 00:09:27,120 IN ADDITION AT THE REAR OF THE 253 00:09:27,120 --> 00:09:29,640 CELL, YOU'VE MORE STABILIZED AND 254 00:09:29,640 --> 00:09:31,720 I WILL SHOW A MOVIE WHERE YOU 255 00:09:31,720 --> 00:09:35,440 SEE MORE STABLE ACTIN KNOWN AS 256 00:09:35,440 --> 00:09:37,760 UTROAPIN AND WHERE IT LOCALIZED 257 00:09:37,760 --> 00:09:39,600 NEAR THE END OF THE CELL, FOR 258 00:09:39,600 --> 00:09:40,440 ROW A ACTIVATION, SO THIS IS 259 00:09:40,440 --> 00:09:41,920 JUST SORT OF A FUN MOVE FREE 260 00:09:41,920 --> 00:09:43,720 RADICALS YEARS AGO THAT IF YOU 261 00:09:43,720 --> 00:09:45,160 ACTIVATE RAC AT THE LEADING 262 00:09:45,160 --> 00:09:47,480 EDGE, WHAT HAPPENS TO MODEL 263 00:09:47,480 --> 00:09:47,920 CITIZEN TILL INVIVO. 264 00:09:47,920 --> 00:09:50,360 SO IN THIS CASE, IF YOU 265 00:09:50,360 --> 00:09:51,560 REPEATEDLY, PHOTO ACTIVATE RAC 266 00:09:51,560 --> 00:09:53,560 AT THE LEADING EDGE, WHAT YOU 267 00:09:53,560 --> 00:09:56,000 CAN DO IS CONTROL WHERE 268 00:09:56,000 --> 00:09:57,680 NEUTROPHILS ARE MIGRATING INVIVO 269 00:09:57,680 --> 00:10:00,160 AND IN THIS CASE SPELL THE WORD 270 00:10:00,160 --> 00:10:00,360 RAC. 271 00:10:00,360 --> 00:10:02,680 SO THIS SHOWS THAT RAC SIGNALING 272 00:10:02,680 --> 00:10:05,720 IS SUFFICIENT TO INDUCE 273 00:10:05,720 --> 00:10:06,840 NEUTROPHIL POLARIZATION AND 274 00:10:06,840 --> 00:10:07,120 MOTILITY. 275 00:10:07,120 --> 00:10:08,480 I WAS TALKING TO CLAIR WATER MAN 276 00:10:08,480 --> 00:10:10,200 EARLIER TODAY ABOUT THIS WHOLE 277 00:10:10,200 --> 00:10:13,440 IDEA OF CELLS BEING ABLE TO GET 278 00:10:13,440 --> 00:10:15,400 THROUGH NARROW CHANNELS INVIVO 279 00:10:15,400 --> 00:10:16,320 AND WITHIN TISSUES AND YOU CAN 280 00:10:16,320 --> 00:10:19,960 SEE HERE WHEN WE REPEATEDLY 281 00:10:19,960 --> 00:10:23,200 PHOTO ACTIVATE RAC, THE LEADING 282 00:10:23,200 --> 00:10:23,800 EDGE, THIS NEUTROPHIL CAN 283 00:10:23,800 --> 00:10:25,120 SQUEEZE THROUGH A VERY NARROW 284 00:10:25,120 --> 00:10:25,320 SPACE. 285 00:10:25,320 --> 00:10:27,960 SO WHAT HAPPENS WITH THE 286 00:10:27,960 --> 00:10:30,800 DOMINANT INHIBITORY MUTATION OF 287 00:10:30,800 --> 00:10:31,840 RAC 2? 288 00:10:31,840 --> 00:10:33,600 SO THIS IS CONTROL NEUTROPHILS 289 00:10:33,600 --> 00:10:35,360 WHICH YOU CAN SEE MIGRATE 290 00:10:35,360 --> 00:10:37,760 RAPIDLY WITH A POLARIZATION OF 291 00:10:37,760 --> 00:10:40,640 STABILIZED OF ACTIN TO THE REAR 292 00:10:40,640 --> 00:10:42,760 AS INDICATED BY THE UTROPIN GFP 293 00:10:42,760 --> 00:10:45,160 AND WHEN YOU HAVE THIS DOMINANT 294 00:10:45,160 --> 00:10:46,720 INHIBITORY MUTATION OF RAC 2, 295 00:10:46,720 --> 00:10:49,000 WHAT YOU SEE IS THE NEUTROPHILS 296 00:10:49,000 --> 00:10:50,360 DON'T MIGRATE, THEY DON'T 297 00:10:50,360 --> 00:10:51,560 POLARIZE AND WHAT'S INTERESTING 298 00:10:51,560 --> 00:10:54,760 IS YOU HAVE STABILIZE OF ACTIN 299 00:10:54,760 --> 00:10:56,600 THROUGHOUT THE CORTICAL REGION 300 00:10:56,600 --> 00:10:58,760 OF THE NEUTROPHIL. 301 00:10:58,760 --> 00:10:59,760 SO SIGNIFICANTLY IMPAIRED 302 00:10:59,760 --> 00:11:00,160 MOTILITY. 303 00:11:00,160 --> 00:11:02,600 AND THE CONSEQUENCE OF THIS IS 304 00:11:02,600 --> 00:11:04,560 THAT THE ZEBRA FISH RAPIDLY DIE 305 00:11:04,560 --> 00:11:06,160 FROM INFECTION AND NEUTROPHILS 306 00:11:06,160 --> 00:11:08,960 ARE NOT RECRUITED TO PSEUDOMOAN 307 00:11:08,960 --> 00:11:11,360 ISOTOPE INJECTED INTO THE EAR, 308 00:11:11,360 --> 00:11:14,160 SO THIS RECAPITULATES THE HUMAN 309 00:11:14,160 --> 00:11:14,400 DISEASE. 310 00:11:14,400 --> 00:11:16,320 WHILE THERE ARE A NUMBER OF 311 00:11:16,320 --> 00:11:17,760 VARIANTS IN RAC 2 THERE ARE 312 00:11:17,760 --> 00:11:18,560 ASSOCIATED OTHER DISEASES, SO 313 00:11:18,560 --> 00:11:19,800 THIS SAY VERY POWERFUL SYSTEM TO 314 00:11:19,800 --> 00:11:23,480 LOOK AT SOME OF THOSE OTHER RAC 315 00:11:23,480 --> 00:11:24,000 2 VARIANTS. 316 00:11:24,000 --> 00:11:26,520 SO AN EXAMPLE WAS PUBLISHED BY 317 00:11:26,520 --> 00:11:29,640 STEVE HOLLAND'S LAB, AND THIS IS 318 00:11:29,640 --> 00:11:32,320 AN ACTIVATING FORM OF RAC 2, 319 00:11:32,320 --> 00:11:34,360 WHICH IS ASSOCIATED WITH IMMUNE 320 00:11:34,360 --> 00:11:37,720 DISREGULATION AND IN THIS CASE, 321 00:11:37,720 --> 00:11:39,160 INFLAMMATION AND TISSUES, BRONCH 322 00:11:39,160 --> 00:11:41,080 YECT SIS AND WHAT YOU CAN SEE IN 323 00:11:41,080 --> 00:11:42,840 THIS CASE IS THAT CONTROL 324 00:11:42,840 --> 00:11:43,680 NEUTROPHILS MIGRATE TO THE SITE 325 00:11:43,680 --> 00:11:47,280 OF DAMAGE AND LEAVE WITH THE 326 00:11:47,280 --> 00:11:49,440 ACTIVATED MUTATION IN RAC 2, YOU 327 00:11:49,440 --> 00:11:51,800 SEE THAT THE NEUTROPHILS MIGRATE 328 00:11:51,800 --> 00:11:53,520 FASTER, QUANTIFIED HERE, AND 329 00:11:53,520 --> 00:11:54,120 MORE NEUTROPHILS ARE RECRUITED 330 00:11:54,120 --> 00:11:55,560 AND THIS IS WHAT YOU WOULD 331 00:11:55,560 --> 00:11:57,680 EXPECT WITH AN INFLAMED TISSUE, 332 00:11:57,680 --> 00:11:59,280 YOU WOULD EXPECT MORE 333 00:11:59,280 --> 00:12:02,000 NEUTROPHILS WOULD BE MIGRATING 334 00:12:02,000 --> 00:12:02,440 THERE. 335 00:12:02,440 --> 00:12:04,720 AND THAT THERE MOTILITY WOULD BE 336 00:12:04,720 --> 00:12:05,840 INCREASED SO WE BECAME VERY 337 00:12:05,840 --> 00:12:08,640 INTERESTED IN THIS CASE 338 00:12:08,640 --> 00:12:11,640 QUESTION, BASED ON PATIENTS THAT 339 00:12:11,640 --> 00:12:14,760 I SAW YEARS AGO, WITH NEONATAL 340 00:12:14,760 --> 00:12:15,680 ONSET MULTISYSTEM INFLAMMATORY 341 00:12:15,680 --> 00:12:20,920 DISEASE, SO I HAD A LOT OF FUN 342 00:12:20,920 --> 00:12:22,520 TALKING TO COLLEAGUES AND 343 00:12:22,520 --> 00:12:25,080 OBVIOUSLY SOME AMAZING WORK HAS 344 00:12:25,080 --> 00:12:28,160 BEEN DONE HERE LOOKING AT 345 00:12:28,160 --> 00:12:28,760 VARIOUS AUTOINFLAMMATORY 346 00:12:28,760 --> 00:12:29,040 DISEASES. 347 00:12:29,040 --> 00:12:30,360 THIS PATIENT AT THE TIME THAT I 348 00:12:30,360 --> 00:12:33,480 SAW HER, WE DIDN'T KNOW WHAT HER 349 00:12:33,480 --> 00:12:35,880 UNDERLYING GENETIC MUTATION WAS. 350 00:12:35,880 --> 00:12:38,600 THAT SHE HAD A DISEASE 351 00:12:38,600 --> 00:12:41,160 CALLEDNYONATAL ONSET MULTISYSTEM 352 00:12:41,160 --> 00:12:42,480 INFLAMMATORY DISEASE AND LATER 353 00:12:42,480 --> 00:12:46,440 WE FOUND SHE HAD AN NLR3 P 354 00:12:46,440 --> 00:12:46,920 MUTATION. 355 00:12:46,920 --> 00:12:49,080 AND SHE PRESENT WIDE FEVER, SKIN 356 00:12:49,080 --> 00:12:50,560 RASH, JOINT SWELLING AND WHEN 357 00:12:50,560 --> 00:12:53,440 YOU BIOPSY HER SKIN, YOU SAW 358 00:12:53,440 --> 00:12:54,520 NEUTROPHIL INFILTRATION INTO THE 359 00:12:54,520 --> 00:12:55,880 SKIN. 360 00:12:55,880 --> 00:12:57,240 BUT WHAT WAS VERY INTERESTING IS 361 00:12:57,240 --> 00:12:59,720 WE LOOKED AT HER NEUTROPHILS AND 362 00:12:59,720 --> 00:13:04,080 WE LOOKEDDA THE NEUTROPHILS OF 363 00:13:04,080 --> 00:13:05,000 SEVERAL PATIENTS WITH NOMID AND 364 00:13:05,000 --> 00:13:07,160 IN ALL CASES THEIR NEUTROPHILS 365 00:13:07,160 --> 00:13:08,440 SHOWED IMPAIRED CHEMO TAXIS TO 366 00:13:08,440 --> 00:13:10,600 ALL THE CHEMO KINES WE STUDY. 367 00:13:10,600 --> 00:13:12,920 SO WHY IS IT THAT SOMEONE WITH A 368 00:13:12,920 --> 00:13:14,600 LOT OF INFLAMMATION AND TISSUES, 369 00:13:14,600 --> 00:13:17,120 WHY WOULD THEIR NEUTROPHILS SHOW 370 00:13:17,120 --> 00:13:17,800 IMPAIRED CHEMO TAXIS. 371 00:13:17,800 --> 00:13:20,040 SO THIS RAISED THE QUESTION THAT 372 00:13:20,040 --> 00:13:23,000 HAS REALLY BEEN A DRIVING FORCE 373 00:13:23,000 --> 00:13:27,160 OF SOME OF OUR RESEARCH IS WHAT 374 00:13:27,160 --> 00:13:27,960 THE RELATIONSHIP BETWEEN 375 00:13:27,960 --> 00:13:31,360 NEUTROPHIL MIGRATION AND 376 00:13:31,360 --> 00:13:31,680 INFLAMMATION? 377 00:13:31,680 --> 00:13:32,640 SO MANY YEARS AGO WE WERE 1 OF 378 00:13:32,640 --> 00:13:35,960 THE FIRST LABS TO GENERATE A 379 00:13:35,960 --> 00:13:37,040 TRANSGENIC THAT LABELS 380 00:13:37,040 --> 00:13:38,560 NEUTROPHILS AND WHEN WE WOUND 381 00:13:38,560 --> 00:13:40,360 THE ZEBRA FISH, WHAT WE FOUND IS 382 00:13:40,360 --> 00:13:42,120 THAT NEUTROPHILS CAME TO THE 383 00:13:42,120 --> 00:13:44,240 SITE OF TISSUE DAMAGE AND THEN 384 00:13:44,240 --> 00:13:47,400 VERY PRIZINGLY, THEY LEFT. 385 00:13:47,400 --> 00:13:50,360 SO AT THE TIME, THE DOGMA WAS 386 00:13:50,360 --> 00:13:52,440 THAT NEUTROPHILS GO INTO SITES 387 00:13:52,440 --> 00:13:53,920 OF TISSUE DAMAGE AND THEY DIE 388 00:13:53,920 --> 00:13:55,560 AND THEN THEY'RE CLEARED BY 389 00:13:55,560 --> 00:13:57,160 MACROPHAGES AND THIS SUGGESTED 390 00:13:57,160 --> 00:13:58,760 SOMETHING VERY DIFFERENT THAT 391 00:13:58,760 --> 00:14:01,960 THEY CAN COME INTO SITES OF 392 00:14:01,960 --> 00:14:03,000 TISSUE DAMAGE AND ACTUALLY 393 00:14:03,000 --> 00:14:03,200 LEAVE. 394 00:14:03,200 --> 00:14:04,240 SO ANOTHER TOOL THAT I AM GOING 395 00:14:04,240 --> 00:14:06,960 TO BRING UP LATER IS 1 WHERE WE 396 00:14:06,960 --> 00:14:08,120 CAN ACTUALLY PHOTO CONVERT 397 00:14:08,120 --> 00:14:09,680 NEUTROPHILS THAT HAVE ARRIVED AT 398 00:14:09,680 --> 00:14:12,000 THE WOUND IN THIS CASE, WE PHOTO 399 00:14:12,000 --> 00:14:13,520 CONVERTED THE FIRST 5 400 00:14:13,520 --> 00:14:15,200 NEUTROPHILS, THAT ARRIVED AT THE 401 00:14:15,200 --> 00:14:19,560 WOUND, AND THEN WE CAN TRAP THE 402 00:14:19,560 --> 00:14:20,440 FATE, AND WHAT WAS INTERESTING 403 00:14:20,440 --> 00:14:23,360 TO LOOK AT THESE FIRST 5 404 00:14:23,360 --> 00:14:25,360 NEUTROPHILS, WITHIN THE TISSUES 405 00:14:25,360 --> 00:14:26,440 THEY MIGRATE RANDOMLY. 406 00:14:26,440 --> 00:14:27,400 THEY ALMOST PONDER BEFORE 407 00:14:27,400 --> 00:14:29,560 FINALLY GOING BACK INTO THE 408 00:14:29,560 --> 00:14:29,880 CIRCULATION. 409 00:14:29,880 --> 00:14:32,160 SO THEN FINALLY THEY WILL 410 00:14:32,160 --> 00:14:32,760 UNDERGO REVERSE TRANSMIGRATION, 411 00:14:32,760 --> 00:14:34,880 GO INTO THE CIRCULATION, AND WE 412 00:14:34,880 --> 00:14:38,920 CAN FIND THEM IN TISSUES 2 DAYS 413 00:14:38,920 --> 00:14:39,240 LATER. 414 00:14:39,240 --> 00:14:43,400 AND SO, THEY WILL GO TO VARIOUS 415 00:14:43,400 --> 00:14:44,440 TISSUES INCLUDING HEMATOPOIETIC 416 00:14:44,440 --> 00:14:44,760 TISSUE. 417 00:14:44,760 --> 00:14:46,640 SO, JUST TO ANSWER THIS 418 00:14:46,640 --> 00:14:49,200 QUESTION, HOW DOES NEUTROPHIL 419 00:14:49,200 --> 00:14:50,320 INFLAMMATION RESOLVE, AND SOME 420 00:14:50,320 --> 00:14:52,760 TYPES OF TISSUE DAMAGE IN 421 00:14:52,760 --> 00:14:53,640 PARTICULAR UNINFECTED TISSUE 422 00:14:53,640 --> 00:14:56,000 DAMAGE, WE THINK THAT PROUDLY 423 00:14:56,000 --> 00:14:57,320 THE PRIMARY MECHANISM RESOLUTION 424 00:14:57,320 --> 00:15:00,160 CAN OFTEN BE THIS NEUTROPHIL 425 00:15:00,160 --> 00:15:01,480 REVERSE MIGRATION, SO RATHER 426 00:15:01,480 --> 00:15:02,840 THAN THIS TRADITIONAL IDEA OF 427 00:15:02,840 --> 00:15:04,760 NEUTROPHILS GOING IN AND DYING 428 00:15:04,760 --> 00:15:06,520 AND BEING CLEAVED BY 429 00:15:06,520 --> 00:15:07,680 MACROPHAGES, THEY WILL GO IN AND 430 00:15:07,680 --> 00:15:10,160 THEY WILL LEAVE BY REVERSE 431 00:15:10,160 --> 00:15:10,440 MIGRATION. 432 00:15:10,440 --> 00:15:13,320 SO THERE WAS A LOT OF SKEPTICISM 433 00:15:13,320 --> 00:15:17,240 ABOUT THIS FOR A LONG TIME WITH 434 00:15:17,240 --> 00:15:19,960 THE ZEBRA FISH SPECIFIC 435 00:15:19,960 --> 00:15:20,280 PHENOMENON? 436 00:15:20,280 --> 00:15:22,800 AND THEN AFTER A DECADE WHEN WE 437 00:15:22,800 --> 00:15:25,200 FIRST MADE THIS OBSERVATION FROM 438 00:15:25,200 --> 00:15:27,440 PAUL COOP'S LAB AM, SHOWED IF 439 00:15:27,440 --> 00:15:29,360 YOU LOOK AT THOSE AND PHOTO 440 00:15:29,360 --> 00:15:30,960 CONVERT THE NEUTROPHILS THEY 441 00:15:30,960 --> 00:15:32,280 LEAVE THE SITE OF TISSUE DAMAGE 442 00:15:32,280 --> 00:15:33,560 AND THEY TRAFFIC TO THE LUNG AND 443 00:15:33,560 --> 00:15:36,880 THEY TRAFFIC TO THE BONE MARROW. 444 00:15:36,880 --> 00:15:38,760 SUPPORTING THE IDEA THAT AGAIN, 445 00:15:38,760 --> 00:15:40,040 NEUTROPHIL REVERSE MIGRATION IS 446 00:15:40,040 --> 00:15:42,400 INVOLVED IN RESOLUTION OF 447 00:15:42,400 --> 00:15:42,760 INFLAMMATION. 448 00:15:42,760 --> 00:15:45,160 BUT THERE'S ALSO BEEN SOME VERY 449 00:15:45,160 --> 00:15:47,680 EXCITING DATA SUGGESTING THAT 450 00:15:47,680 --> 00:15:49,240 NEUTROPHIL REVERSE MIGRATION MAY 451 00:15:49,240 --> 00:15:51,160 ALSO BE INVOLVED IN SYSTEMIC 452 00:15:51,160 --> 00:15:53,720 INFLAMMATION, MAY HAVE DISEASE 453 00:15:53,720 --> 00:15:55,880 IMPLICATIONS RELATED FOR EXAMPLE 454 00:15:55,880 --> 00:15:58,760 TO PAN KRE TINSA TITIS AND THEN 455 00:15:58,760 --> 00:16:00,560 ARDS OR IN THE CASE OF 456 00:16:00,560 --> 00:16:02,560 METASTASIS WHERE TUMOR CELLS MAY 457 00:16:02,560 --> 00:16:05,280 BE ASSOCIATE WIDE NEUTROPHILS IN 458 00:16:05,280 --> 00:16:05,800 THE CIRCULATION. 459 00:16:05,800 --> 00:16:07,040 SO THESE LOCATIONS MAY BE BROAD, 460 00:16:07,040 --> 00:16:09,560 WE HAVE DONE A LITTLE BIT OF 461 00:16:09,560 --> 00:16:12,160 WORK LOOKING AT ZEBRA FISH 462 00:16:12,160 --> 00:16:13,680 MODELS OF INFLAMMATION, WHERE WE 463 00:16:13,680 --> 00:16:16,040 FEED ZEBRA FISH NORMAL OR HIGH 464 00:16:16,040 --> 00:16:17,600 FAT DIET AND THEN WE LOOK AT 465 00:16:17,600 --> 00:16:18,960 NEUTROPHIL RECRUITMENT AND YOU 466 00:16:18,960 --> 00:16:21,560 CAN SEE WHERE THE HIGH FAT DIET 467 00:16:21,560 --> 00:16:22,760 IS KIND OF SCARE SKPE BAKUGAN 468 00:16:22,760 --> 00:16:24,040 LOTS OF NEUTROPHILS ARE 469 00:16:24,040 --> 00:16:25,120 RECRUITED TO THE WOUND AND THIS 470 00:16:25,120 --> 00:16:26,960 WORK IS BEING DONE AND PURSUED 471 00:16:26,960 --> 00:16:29,760 NOW IN THE LABORATORY OF A 472 00:16:29,760 --> 00:16:33,880 FORMER POST DOC, SOFIA 473 00:16:33,880 --> 00:16:38,240 DE OLIVEIRA, WHO IS A PROFESSOR 474 00:16:38,240 --> 00:16:39,600 AT ALBERT EINSTEIN. 475 00:16:39,600 --> 00:16:40,880 SO SOFIA ASKED THE QUESTION 476 00:16:40,880 --> 00:16:45,160 WHERE ARE THEY COMING FROM? 477 00:16:45,160 --> 00:16:46,240 OUT OF CIRCULATION? 478 00:16:46,240 --> 00:16:47,160 HEMATOPOIETIC TISSUE? 479 00:16:47,160 --> 00:16:47,880 SO SHE CONVERTED NEUTROPHILS 480 00:16:47,880 --> 00:16:51,320 FROM THE LIVER AND FOUND THAT 481 00:16:51,320 --> 00:16:53,680 THE--IN THE CASE OF A HIFAT DIET 482 00:16:53,680 --> 00:16:54,920 MANY OF THESE NEUTROPHILS WERE 483 00:16:54,920 --> 00:16:55,800 ACTUALLY COMING FROM THE LIVER. 484 00:16:55,800 --> 00:16:58,760 THIS WAS NOT THE CASE WITH 485 00:16:58,760 --> 00:16:59,160 CONTROL. 486 00:16:59,160 --> 00:17:02,240 SO AGAIN, THIS SUPPORTS THE IDEA 487 00:17:02,240 --> 00:17:03,760 THAT NEUTROPHIL WILL REVERSE 488 00:17:03,760 --> 00:17:05,520 MIGRATION AND THAT CASE THAT THE 489 00:17:05,520 --> 00:17:08,040 LIVER WILL CONTRIBUTE TO 490 00:17:08,040 --> 00:17:09,680 SYSTEMIC INFLAMMATORY RESPONSE. 491 00:17:09,680 --> 00:17:20,320 SO, WE AND OTHERS HAVE BEEN 492 00:17:20,320 --> 00:17:23,160 INTERESTED IN WHAT REGULATES 493 00:17:23,160 --> 00:17:23,920 NEUTROPHIL REVERSE MIGRATION? 494 00:17:23,920 --> 00:17:26,800 SO WHEEF BEEN LOOKING AT CHEMO 495 00:17:26,800 --> 00:17:27,960 KINES RECEPTOR SIGNALING, NOW WE 496 00:17:27,960 --> 00:17:29,400 KNOW THAT CHEMO KINES ARE VERY 497 00:17:29,400 --> 00:17:30,560 IMPORTANT FOR THE ONSET OF 498 00:17:30,560 --> 00:17:33,720 INFLAMMATION, SO IF YOU BLOCK 499 00:17:33,720 --> 00:17:34,480 IL8 SIGNALING, NEUTROPHIL 500 00:17:34,480 --> 00:17:38,960 RECRUITMENT TO WOUND CAN BE 501 00:17:38,960 --> 00:17:39,280 IMPAIRED. 502 00:17:39,280 --> 00:17:40,920 BUT 1 THING THAT WAS INTERESTING 503 00:17:40,920 --> 00:17:42,800 TO US IS WHEN WE CONTROLLER OF 504 00:17:42,800 --> 00:17:44,760 BANKED OUT 1 OF THE ISOFORMS IN 505 00:17:44,760 --> 00:17:48,200 ZEBRA FISH AND 1 OF THE 506 00:17:48,200 --> 00:17:49,600 RECEPTORS CXCRC2 IN ZEBRA FISH, 507 00:17:49,600 --> 00:17:51,240 WE FOUND THAT NEUTROPHILS WERE 508 00:17:51,240 --> 00:17:52,400 MORE PERSISTENT AT THE WOUND. 509 00:17:52,400 --> 00:17:54,320 THEY DID NOT HAVE THIS RANDOM 510 00:17:54,320 --> 00:18:00,160 MOTILITY AND THEY HAD IMPAIRED 511 00:18:00,160 --> 00:18:01,760 REVERSE MIGRATION. 512 00:18:01,760 --> 00:18:04,560 NOW WORK DONE BY [INDISCERNIBLE] 513 00:18:04,560 --> 00:18:06,560 GROUP AT CAMBRIDGE HAS GONE ON 514 00:18:06,560 --> 00:18:08,240 ON TO SHOW NICELY THAT WHEN THE 515 00:18:08,240 --> 00:18:09,680 RECEPTOR IS STAINED ON THE CELL 516 00:18:09,680 --> 00:18:10,440 SURFACE, THIS IS ASSOCIATE WIDE 517 00:18:10,440 --> 00:18:11,840 A ROLE IN THE VERMEN INFECTED 518 00:18:11,840 --> 00:18:14,560 VERSE MIGRATION, WHEN IT GETS 519 00:18:14,560 --> 00:18:16,200 INTERNALIZED THEN THE 520 00:18:16,200 --> 00:18:17,320 NEUTROPHILS, THAT'S NOT AS 521 00:18:17,320 --> 00:18:21,680 INVOLVED IN THE REVERSE 522 00:18:21,680 --> 00:18:22,160 MIGRATION PROCESS. 523 00:18:22,160 --> 00:18:25,520 SO SUGGESTED TO US THAT THIS 524 00:18:25,520 --> 00:18:28,520 CHEMO KINESIS THAT'S INDUCED FOR 525 00:18:28,520 --> 00:18:29,560 SIGNATURES SIGNALING FOR THE 526 00:18:29,560 --> 00:18:31,960 RECEPTORS IS A STEP IN THE 527 00:18:31,960 --> 00:18:33,640 CASCADE. 528 00:18:33,640 --> 00:18:35,160 IT ALSO RAISES QUESTIONS ABOUT A 529 00:18:35,160 --> 00:18:36,760 PATHWAY THAT IS TARGETED 530 00:18:36,760 --> 00:18:37,880 THERAPEUTICALLY THAT MAY IN FACT 531 00:18:37,880 --> 00:18:40,680 LEAD IN SOME CASES TO LESS 532 00:18:40,680 --> 00:18:41,640 INFLAMMATION AND OTHERS 533 00:18:41,640 --> 00:18:42,680 POTENTIALLY TO MORE INFLAMMATION 534 00:18:42,680 --> 00:18:45,360 WHICH WE KNOW IS TRUE OF A 535 00:18:45,360 --> 00:18:49,160 NUMBER OF DIFFERENT TARGETS. 536 00:18:49,160 --> 00:18:51,080 SO THIS GETS BACK TO THIS IDEA, 537 00:18:51,080 --> 00:18:53,640 WE'RE INTERESTED IN THIS WHOLE 538 00:18:53,640 --> 00:18:54,360 BALANCE AND NEUTROPHIL 539 00:18:54,360 --> 00:18:59,760 INFLAMMATION AND TISSUE REPAIR. 540 00:18:59,760 --> 00:19:03,080 IF YOU HAVE TOO MUCH 541 00:19:03,080 --> 00:19:03,800 INFLAMMATION, THEY DEPLETE. 542 00:19:03,800 --> 00:19:06,520 SO IF WE DEPLETE THESE, WE FOUND 543 00:19:06,520 --> 00:19:08,320 MANY MODELS TISSUE REPAIR FASTER 544 00:19:08,320 --> 00:19:09,960 BUT NOT ALL NEUTROPHILS ARE THE 545 00:19:09,960 --> 00:19:10,160 SAME. 546 00:19:10,160 --> 00:19:11,800 A RECENT PAPER IN IMMUNOLOGY 547 00:19:11,800 --> 00:19:14,720 FOUND THAT IF YOU DEPLETE 548 00:19:14,720 --> 00:19:16,360 NEUTROPHILS THEY'RE REQUIRED FOR 549 00:19:16,360 --> 00:19:20,240 EFFICIENT REPAIR BY REMODELING 550 00:19:20,240 --> 00:19:22,720 THE MATRIX, SO, ALL NEUTROPHILS 551 00:19:22,720 --> 00:19:23,280 ARE NOT ALIKE. 552 00:19:23,280 --> 00:19:25,720 OBVIOUSLY IF YOU HAVE 2 FEW 553 00:19:25,720 --> 00:19:27,560 NEUTROPHILS YOU END UP WITH 554 00:19:27,560 --> 00:19:29,400 IMMUNE DEFICIENCY AND IN THAT 555 00:19:29,400 --> 00:19:34,160 CASE, CHRONIC INFECTION AND 556 00:19:34,160 --> 00:19:34,760 IMPAIRED HEALING. 557 00:19:34,760 --> 00:19:36,960 SO WE WERE INTERESTED IN 558 00:19:36,960 --> 00:19:39,320 IDENTIFYING NEW MECHANISMS THAT 559 00:19:39,320 --> 00:19:40,960 MIGHT REGULATE INFLAMMATION 560 00:19:40,960 --> 00:19:41,280 RESOLUTION. 561 00:19:41,280 --> 00:19:43,520 AND FOR THIS, WE WANTED TO LOOK 562 00:19:43,520 --> 00:19:45,680 FOR FACTORS THAT WERE 563 00:19:45,680 --> 00:19:46,640 UPREGULATED AND STERILE 564 00:19:46,640 --> 00:19:48,560 TRIGGERS, BUT NOT INFECTION. 565 00:19:48,560 --> 00:19:52,480 AND TO DO THIS WORK, WE TOOK 566 00:19:52,480 --> 00:19:54,960 ADVANTAGE OF 2 DIFFERENT 567 00:19:54,960 --> 00:19:55,960 INFLAMMATION MODELS, ACUTE 568 00:19:55,960 --> 00:19:57,640 WOUNDING OF THE ZEBRA FISH WHERE 569 00:19:57,640 --> 00:19:59,360 YOU GET RECRUITMENT OF 570 00:19:59,360 --> 00:20:01,000 NEUTROPHILS AND MACROPHAGES AND 571 00:20:01,000 --> 00:20:03,160 THEN, CANCER AS KIND OF AN 572 00:20:03,160 --> 00:20:04,040 UNHEALED WOUND. 573 00:20:04,040 --> 00:20:06,560 WE HAVE CHRONIC ASSOCIATION OF 574 00:20:06,560 --> 00:20:08,640 NEUTROPHILS AND MACK RAPHAGES 575 00:20:08,640 --> 00:20:09,240 WITH TRANSFORM CELLS. 576 00:20:09,240 --> 00:20:11,680 AND WHAT WE DID IS WE DID 577 00:20:11,680 --> 00:20:17,040 SOMETHING CALLED TRAP SEQ, 578 00:20:17,040 --> 00:20:20,200 RNA-SEQ AND THEN TARGETED 579 00:20:20,200 --> 00:20:20,640 PATHWAYSA BY CRISPR. 580 00:20:20,640 --> 00:20:25,000 SO TO DO THIS, WE GENERATED 581 00:20:25,000 --> 00:20:26,880 ZEBRA FISH LINES THAT EXPRESS 582 00:20:26,880 --> 00:20:29,320 THE RIBOSOMAL L10 A PROTEIN FUSE 583 00:20:29,320 --> 00:20:32,080 TO GFP AND WE MADE LINES AND 584 00:20:32,080 --> 00:20:33,040 NEUTROPHILS, MACROPHAGES AND 585 00:20:33,040 --> 00:20:33,920 EPITHELIAL CELLS. 586 00:20:33,920 --> 00:20:36,720 AND THEN, IN THIS CASE, WORKED 587 00:20:36,720 --> 00:20:39,360 ON BY RUTH HOUSRIGHT A FORMER 588 00:20:39,360 --> 00:20:41,600 GRADUATE STUDENT IN THE LAB, SHE 589 00:20:41,600 --> 00:20:43,240 INJURED THE FISH IN MULTIPLE 590 00:20:43,240 --> 00:20:46,320 PLACES AND THEN 3 HOURS POST 591 00:20:46,320 --> 00:20:50,560 WOUND COLLECTED THE SAMPLES FOR 592 00:20:50,560 --> 00:20:52,120 THIS TRAP-SEQ AND THERE WAS A 593 00:20:52,120 --> 00:20:55,360 GENE THAT WAS UPREGULATED BOTH 594 00:20:55,360 --> 00:20:56,000 IN NEUTROPHILS AND MACROPHAGES, 595 00:20:56,000 --> 00:20:57,120 1 THING TO NOTE IS SOMETHING 596 00:20:57,120 --> 00:20:59,160 THAT WAS INTERESTING IS VERY FEW 597 00:20:59,160 --> 00:21:00,400 GENES WERE COMMONLY UPREGULATED 598 00:21:00,400 --> 00:21:02,320 IN A DIFFERENT CELL TYPES BUT 599 00:21:02,320 --> 00:21:06,440 THERE WAS 1 THAT WAS UPRIGGING 600 00:21:06,440 --> 00:21:07,880 BOTH NEUTROPHILS AND MACROPHAGES 601 00:21:07,880 --> 00:21:10,400 WITH STERILE INJURY AND THAT WAS 602 00:21:10,400 --> 00:21:11,760 A GENE CALLED MYELOID DERIVED 603 00:21:11,760 --> 00:21:12,800 GROWTH FACTOR AND AT THAT TIME 604 00:21:12,800 --> 00:21:14,320 THERE WAS NOT MUCH OF IT UNKNOWN 605 00:21:14,320 --> 00:21:16,960 ABOUT IT IN THE LITERATURE, SO 606 00:21:16,960 --> 00:21:21,440 MYELOID DRIVE GROWTH FACTOR IS A 607 00:21:21,440 --> 00:21:22,560 PERIKRIN 17 KILODALTON SECRETED 608 00:21:22,560 --> 00:21:26,320 PROTEIN, IT WAS REFERRED TO AS A 609 00:21:26,320 --> 00:21:29,480 SPECIFIC ORF THAT WAS FOUND TO 610 00:21:29,480 --> 00:21:32,240 BE UPREGULATED CAN IN FLUM 611 00:21:32,240 --> 00:21:33,000 INFLAMMATION, SYNOVIAL FLUID 612 00:21:33,000 --> 00:21:35,640 FROM OUR RA PATIENTS AND 613 00:21:35,640 --> 00:21:36,960 HEPATITIS EATIC CARCINOMA THAT 614 00:21:36,960 --> 00:21:38,360 WAS ASSOCIATE WIDE POOR 615 00:21:38,360 --> 00:21:44,160 PROCEEDINGINOSEIS SO THEN IT WAS 616 00:21:44,160 --> 00:21:45,400 NAMED MYELOID-DERIVED GROWTH 617 00:21:45,400 --> 00:21:47,680 FACTOR, THAT WAS SECRETED BY 618 00:21:47,680 --> 00:21:49,360 MONOCYTES AND MACROPHAGES AND 619 00:21:49,360 --> 00:21:52,880 TISSUE REPAIR IN A MURINE MODEL 620 00:21:52,880 --> 00:21:54,400 OF MY O CARDIAL INFARCTION. 621 00:21:54,400 --> 00:21:56,920 AND THIS KNOCK OUT MOUSE WAS 622 00:21:56,920 --> 00:21:58,160 PERFECTLY FINE BUT THEY DID MUCH 623 00:21:58,160 --> 00:22:00,640 LESS WELL WHEN THEY HAD AN MI. 624 00:22:00,640 --> 00:22:01,760 SO IT'S AN INTERESTING PROTEIN 625 00:22:01,760 --> 00:22:03,120 THAT WE DON'T KNOW MUCH ABOUT 626 00:22:03,120 --> 00:22:04,560 AND I WILL TELL YOU ABOUT HOW 627 00:22:04,560 --> 00:22:06,880 WE'RE TRYING TO GET AT HOW THIS 628 00:22:06,880 --> 00:22:10,520 IN FACT, WORKS BUT WE STILL HAVE 629 00:22:10,520 --> 00:22:11,200 MANY QUESTIONS REMAINING. 630 00:22:11,200 --> 00:22:13,600 IT CAN BE A RESIDENT ER PROTEIN, 631 00:22:13,600 --> 00:22:21,680 IT CAN ALSO BE SECRETED AND IT 632 00:22:21,680 --> 00:22:24,640 BINDS TO KDEL RECEPTORS, SO IT 633 00:22:24,640 --> 00:22:28,320 HAS A KDEL MOTIF AND IT'S 634 00:22:28,320 --> 00:22:30,120 INCREASED HYPOXIA AND IT'S 635 00:22:30,120 --> 00:22:31,560 CONTINUATION OF DIABETIC KIDNEY 636 00:22:31,560 --> 00:22:33,360 DISEASE AND VASCULAR 637 00:22:33,360 --> 00:22:33,720 INFLAMMATION. 638 00:22:33,720 --> 00:22:36,040 SO WE KNOCKED IT OUT, OF ZEBRA 639 00:22:36,040 --> 00:22:38,520 FISH AND YOU CAN SEE HERE, YOU 640 00:22:38,520 --> 00:22:41,440 GENERATED AN ANTIBODY, AND MADE 641 00:22:41,440 --> 00:22:42,760 A CRISPR MUTANT AND YOU CAN SEE 642 00:22:42,760 --> 00:22:43,920 THAT THE PROTEIN IS GONE. 643 00:22:43,920 --> 00:22:49,400 I JUST WANT TO HIGHLIGHT THAT 644 00:22:49,400 --> 00:22:51,400 THE ZEBRA FISH GROWTH FACTOR IS 645 00:22:51,400 --> 00:22:52,960 HIGHLY CONSERVED BOTH 646 00:22:52,960 --> 00:22:55,560 STRUCTURALLY AND BASED ON AMINO 647 00:22:55,560 --> 00:22:55,960 ACID SIMILARITY. 648 00:22:55,960 --> 00:22:58,160 AND WHAT HAPPENS WHEN WE KNOCK 649 00:22:58,160 --> 00:22:59,040 IT OUT? 650 00:22:59,040 --> 00:23:02,520 SO, THE MODEL THAT I'M GOING TO 651 00:23:02,520 --> 00:23:04,360 BE USING QUITE OFTEN IN THIS 652 00:23:04,360 --> 00:23:05,880 TALK THAT I WANT TO JUST TELL 653 00:23:05,880 --> 00:23:07,760 YOU ABOUT IS A DIFFERENT TYPE OF 654 00:23:07,760 --> 00:23:11,560 INJURY MODEL, IT'S A MORE 655 00:23:11,560 --> 00:23:14,880 SIGNIFICANT INJURY WHERE WE USE 656 00:23:14,880 --> 00:23:17,840 A HEAT SOURCE, A CAUTERY WIRE 657 00:23:17,840 --> 00:23:20,720 AND WE BURN THE TAIL AND THEN WE 658 00:23:20,720 --> 00:23:21,680 LOOK AT INFLAMMATORY RESPONSE 659 00:23:21,680 --> 00:23:22,760 WHICH IS VERY DIFFERENT THAN IF 660 00:23:22,760 --> 00:23:24,560 WE CUT OFF THE TAIL AND I WILL 661 00:23:24,560 --> 00:23:26,360 GET INTO THAT A BIT MORE LATER. 662 00:23:26,360 --> 00:23:29,680 SO LAWN MOWER WE FOUND IN BOTH 663 00:23:29,680 --> 00:23:30,560 THE TAIL TRANSSECTION, WHERE WE 664 00:23:30,560 --> 00:23:33,000 CUT OFF THE TAIL OR WE BIRN THE 665 00:23:33,000 --> 00:23:34,920 ZEBRA FISH TAIL, WE FOUND THAT 666 00:23:34,920 --> 00:23:36,000 DEPLETION OF MYELOID GROWTH 667 00:23:36,000 --> 00:23:37,680 FACTOR LED TO INCREASED 668 00:23:37,680 --> 00:23:41,040 NEUTROPHILS AT THE WOUNDS 669 00:23:41,040 --> 00:23:41,760 QUANTIFIED HERE EVEN 24 HOURS, 670 00:23:41,760 --> 00:23:42,000 POST BURN. 671 00:23:42,000 --> 00:23:44,840 YOU CAN SEE THE NEUTROPHILS 672 00:23:44,840 --> 00:23:45,360 HERE. 673 00:23:45,360 --> 00:23:47,760 IF YOU LOOK AT A MOVIE, WHAT YOU 674 00:23:47,760 --> 00:23:50,760 CAN SEE, IS THERE'S A ROBUST 675 00:23:50,760 --> 00:23:53,000 RESPONSE OF BOTH NEUTROPHILS AND 676 00:23:53,000 --> 00:23:53,960 MACROPHAGES TO A BURN WOUND AND 677 00:23:53,960 --> 00:23:55,360 IF YOU LOOK AT THE SIDE VIEW 678 00:23:55,360 --> 00:23:58,160 HERE, CAN YOU SEE EVEN MORE THAT 679 00:23:58,160 --> 00:24:00,200 WHEN YOU KNOCK OUT MILEN GROWTH 680 00:24:00,200 --> 00:24:01,920 FACTOR, THERE ARE EVEN MORE 681 00:24:01,920 --> 00:24:03,480 NEUTROPHILS RECRUITED TO A BURN 682 00:24:03,480 --> 00:24:03,680 WOUND. 683 00:24:03,680 --> 00:24:07,360 BUT WE DID NOT SEE A DIFFERENCE 684 00:24:07,360 --> 00:24:08,080 IN MACROPHAGE RECRUITMENT. 685 00:24:08,080 --> 00:24:09,880 SO THIS SUGGESTS THAT MYELOID 686 00:24:09,880 --> 00:24:11,840 DRIVE GROWTH FACTOR IS 687 00:24:11,840 --> 00:24:13,160 NEGATIVELY REGULATING NEUTROPHIL 688 00:24:13,160 --> 00:24:15,480 INFLAMMATION, SO IS THE PROTEIN 689 00:24:15,480 --> 00:24:18,640 SUFFICIENT TO ACTUALLY INHIBIT 690 00:24:18,640 --> 00:24:19,160 NEUTROPHIL INFLAMMATION? 691 00:24:19,160 --> 00:24:21,320 SO WHAT WE DID IS WE TOOK 692 00:24:21,320 --> 00:24:22,280 MYELOID DRIVE GROWTH FACTOR AND 693 00:24:22,280 --> 00:24:24,080 WE INJEKED INTO INTO THE EAR OF 694 00:24:24,080 --> 00:24:26,440 THE ZEBRA FISH, AND WE USED 695 00:24:26,440 --> 00:24:27,960 EITHER CONTROL OR MYELOID 696 00:24:27,960 --> 00:24:29,320 DERIVED GROWTH FACTOR AND WHAT 697 00:24:29,320 --> 00:24:31,600 WE FOUND IS JUST WHEN YOU HIT 698 00:24:31,600 --> 00:24:33,560 THE EAR WITH THE NEEDLE, YOU SEE 699 00:24:33,560 --> 00:24:35,560 NEUTROPHILS COME TO THE EAR SITE 700 00:24:35,560 --> 00:24:38,800 AS A WOUND AND THEN MYELOID 701 00:24:38,800 --> 00:24:40,840 DERIVED GROWTH FACTOR 702 00:24:40,840 --> 00:24:41,840 DRAMATICALLY INHIBIT THE 703 00:24:41,840 --> 00:24:42,760 NEUTROPHIL RECRUITMENT, YOU CAN 704 00:24:42,760 --> 00:24:43,760 SEE THIS QUANTIFIED HERE AND 705 00:24:43,760 --> 00:24:46,080 WHILE WE LOOK AT THE MUTANT AS 706 00:24:46,080 --> 00:24:47,920 COMPARED TO THE NONMUTANT, CAN 707 00:24:47,920 --> 00:24:49,560 YOU SEE, MANY MORE NEUTROPHILS 708 00:24:49,560 --> 00:24:52,200 ARE RECRUITED TO THIS WOUND, AND 709 00:24:52,200 --> 00:24:55,760 THIS IS RESCUED BY INJECTING THE 710 00:24:55,760 --> 00:24:56,480 MYELOID DRIVE GROWTH FACTOR. 711 00:24:56,480 --> 00:24:58,720 SO THIS SUGGESTS THAT THIS IS A 712 00:24:58,720 --> 00:25:00,560 NEW INHIBITOR OF NEUTROPHIL 713 00:25:00,560 --> 00:25:02,120 INFLAMMATION DISP WE'RE REALLY 714 00:25:02,120 --> 00:25:02,760 EXCITED ABOUT IT BECAUSE THERE 715 00:25:02,760 --> 00:25:05,400 AREN'T A LOT OF THINGS THAT 716 00:25:05,400 --> 00:25:06,600 INHIBIT NEUTROPHIL INFLAMMATION 717 00:25:06,600 --> 00:25:08,520 IN RESPONSE TO STERILE INJURY 718 00:25:08,520 --> 00:25:10,000 BUT NOT INFECTION AND THAT'S 719 00:25:10,000 --> 00:25:10,600 WHAT WE FOUND. 720 00:25:10,600 --> 00:25:12,160 SO THIS IS PRETTY COOL. 721 00:25:12,160 --> 00:25:16,400 IT'S SUGGESTS IT MIGHT BE AN 722 00:25:16,400 --> 00:25:17,040 INTERESTING THERAPEUTIC TARGET. 723 00:25:17,040 --> 00:25:19,440 SO HOW DOES MYELOID DRIVE GROWTH 724 00:25:19,440 --> 00:25:20,760 FACTOR AFFECT WOUND HEALING, WE 725 00:25:20,760 --> 00:25:23,000 KNOW IT PLAYS A ROLE IN MI 726 00:25:23,000 --> 00:25:26,640 RESPONSE AND WHAT WE FOUND IS IF 727 00:25:26,640 --> 00:25:29,280 YOU DO A BURN WOUND IN ZEBRA 728 00:25:29,280 --> 00:25:31,600 FISH OR A TAIL TRANSSECTION, YOU 729 00:25:31,600 --> 00:25:33,120 HAVE IMPAIRED, REPAIR AT ALL 730 00:25:33,120 --> 00:25:35,560 TIME POINTS IN THE MYELOID 731 00:25:35,560 --> 00:25:37,360 DERIVED GROWTH FACTOR MUTANT BUT 732 00:25:37,360 --> 00:25:40,560 THE THIN IS GROWING JUST FINE 733 00:25:40,560 --> 00:25:41,040 NONWOUNDED ZEBRA FISH. 734 00:25:41,040 --> 00:25:42,720 AND THEN THE QUESTION IS LOKAY, 735 00:25:42,720 --> 00:25:45,680 SO WE HAVE MORE NEUTROPHILS 736 00:25:45,680 --> 00:25:48,040 THERE, THERE'S IMPAIRED HEALING 737 00:25:48,040 --> 00:25:50,360 OF THE WOUND, ARE NEUTROPHILS 738 00:25:50,360 --> 00:25:50,920 INVOLVED IN THIS PHENOTYPE? 739 00:25:50,920 --> 00:25:53,200 SO WHAT WE DID IS WE LOOKED AT 740 00:25:53,200 --> 00:25:57,760 THE WOUND HEALING DEFECT AND 741 00:25:57,760 --> 00:25:59,000 NEUTROPHIL DEFICIENT ZEBRA FISH. 742 00:25:59,000 --> 00:26:03,440 SO WE USE THAT RAC 2 D57 INLINE 743 00:26:03,440 --> 00:26:08,840 I DESCRIBED BEFORE AND WE DID 744 00:26:08,840 --> 00:26:12,680 MORPHOLIN O OR MYELOID DRIVEN 745 00:26:12,680 --> 00:26:15,640 AND WE SAW THE TRANSIENT GROWTH 746 00:26:15,640 --> 00:26:17,480 MYELOID GROWTH FACTOR BUT THIS 747 00:26:17,480 --> 00:26:20,640 DOES NOT OCCUR IN THE RAC 748 00:26:20,640 --> 00:26:21,520 DEFICIENT ZEBRA FISH. 749 00:26:21,520 --> 00:26:23,480 SO IT SUGGESTS THAT THE WOUND 750 00:26:23,480 --> 00:26:26,800 HEALING DEFECT IS NEUTROPHIL 751 00:26:26,800 --> 00:26:30,480 DEPENDENT IN MYELOID DEFICIENT 752 00:26:30,480 --> 00:26:30,680 LARVA. 753 00:26:30,680 --> 00:26:31,720 SO THEN WE'RE INTERESTED IN THE 754 00:26:31,720 --> 00:26:33,560 QUESTION, WHAT IS THE MECHANISM 755 00:26:33,560 --> 00:26:36,040 OF INCREASED NEUTROPHIL 756 00:26:36,040 --> 00:26:38,320 INFLAMMATION IN MUTANTS, MORE 757 00:26:38,320 --> 00:26:38,960 RECRUITMENT, DIFFERENT IN 758 00:26:38,960 --> 00:26:40,040 REVERSE MIGRATION AND FOR THIS 759 00:26:40,040 --> 00:26:41,440 WE DID THAT PHOTO CONVERSION 760 00:26:41,440 --> 00:26:43,760 EXPERIMENT THAT I SHOWED YOU 761 00:26:43,760 --> 00:26:45,440 EARLIER, WHERE WE PHOTO CONVERT 762 00:26:45,440 --> 00:26:47,160 NEUTROPHILS AT 2 HOURS POST 763 00:26:47,160 --> 00:26:49,360 WOUND AND A TAIL TRANSSECTION, 764 00:26:49,360 --> 00:26:51,560 AND IMAGE AGAIN AT 6 HOURS POST 765 00:26:51,560 --> 00:26:56,600 WOUND, AND YOU CAN SEE HERE THAT 766 00:26:56,600 --> 00:26:58,520 YOU HAVE MANY MORE NEUTROPHILS 767 00:26:58,520 --> 00:26:59,760 IN THE MYELOID DEFICIENT LARVA 768 00:26:59,760 --> 00:27:02,080 AND THAT THE PHOTO CONVERTED 769 00:27:02,080 --> 00:27:03,240 NEUTROPHILS ARE PERSISTENT AND 770 00:27:03,240 --> 00:27:05,080 THIS IS SHOWN HERE. 771 00:27:05,080 --> 00:27:06,360 THERE'S IMPAIRED RESOLUTION OF 772 00:27:06,360 --> 00:27:11,720 THE NEUTROPHILS FROM THIS SITE 773 00:27:11,720 --> 00:27:12,000 OF DAMAGE. 774 00:27:12,000 --> 00:27:13,600 NOW THIS IS ALSO TRUE WHEN WE 775 00:27:13,600 --> 00:27:16,360 LOOK AT THERMAL OR BURN INJURY. 776 00:27:16,360 --> 00:27:17,720 WE THROW A PROTOCONVERTED LATER 777 00:27:17,720 --> 00:27:19,960 BECAUSE THERE'S MUCH MORE 778 00:27:19,960 --> 00:27:21,160 INFLAMMATION AT 4 HOURS AND 779 00:27:21,160 --> 00:27:23,480 IMAGE AT 24 HOURS AND YOU CAN 780 00:27:23,480 --> 00:27:25,720 SEE AGAIN RESOLUTION IS IMPAIRED 781 00:27:25,720 --> 00:27:27,920 IN THE MYELOID DERIVED GROWTH 782 00:27:27,920 --> 00:27:28,400 FACTOR MUTANT. 783 00:27:28,400 --> 00:27:32,080 SO WHAT ELSE IS KNOWN ABOUT 784 00:27:32,080 --> 00:27:33,520 NEUTROPHIL REVERSE MIGRATION AND 785 00:27:33,520 --> 00:27:35,120 SIGNALING PATHWAYS THAT ARE 786 00:27:35,120 --> 00:27:36,080 INVOLVED WITH IT? 787 00:27:36,080 --> 00:27:39,600 AND 1 THAT HAS BEEN IMPLICATED 788 00:27:39,600 --> 00:27:41,760 IS THE HIP 1 ALPHA SO AS I 789 00:27:41,760 --> 00:27:43,160 MENTIONED BOTH GROWTH FACTOR AND 790 00:27:43,160 --> 00:27:47,200 HIP 1 ALPHA ARE UPREGULATED WITH 791 00:27:47,200 --> 00:27:47,640 HYPOXIA. 792 00:27:47,640 --> 00:27:50,520 SO WORK FROM SPHEEF RENSHAH'S 793 00:27:50,520 --> 00:27:53,240 GROUP HAS SHOWN THAT IN HYPOXIA 794 00:27:53,240 --> 00:27:55,280 YOU GET AN UPREGULATION OF HIP 1 795 00:27:55,280 --> 00:27:57,440 ALPHA AND THIS LEADS TO 796 00:27:57,440 --> 00:27:58,160 NEUTROPHIL PERSISTENCE AT THE 797 00:27:58,160 --> 00:27:59,960 WOUND AND THEY DON'T REVERSE 798 00:27:59,960 --> 00:28:01,000 MIGRATE AS MUCH, THEY DON'T DIE 799 00:28:01,000 --> 00:28:03,400 AS MUCH AND TO READ OUT HIP 1 800 00:28:03,400 --> 00:28:09,160 ALPHA SIGNALING, WE CAN LOOK AT 801 00:28:09,160 --> 00:28:11,040 Ph.D. EXPRESSION BY RTPC R, 802 00:28:11,040 --> 00:28:13,720 AND WE FOUND IT WAS INCREASED IN 803 00:28:13,720 --> 00:28:15,640 BOTH TAIL SECTION AND BURN AND 804 00:28:15,640 --> 00:28:16,720 THE MYELOID DRIVE DEFICIENT 805 00:28:16,720 --> 00:28:16,920 FISH. 806 00:28:16,920 --> 00:28:20,240 SO THEN THE QUESTION IS CAN WE 807 00:28:20,240 --> 00:28:21,320 ACTUALLY RESCUE THIS PHENOTYPE 808 00:28:21,320 --> 00:28:24,680 BY KNOCKING DOWN THE HIF 1 HALFA 809 00:28:24,680 --> 00:28:28,960 PATHWAY, SO WE DID THIS, BY 810 00:28:28,960 --> 00:28:30,560 DEPLETING ART 1 USING A 811 00:28:30,560 --> 00:28:34,240 MORPHOLIN O IN EITHER CONTROL OR 812 00:28:34,240 --> 00:28:35,360 THE MYELOID DERIVED DEFICIENT 813 00:28:35,360 --> 00:28:37,440 ZEBRA FISH AND YOU CAN SEE THAT 814 00:28:37,440 --> 00:28:40,840 YOU RESCUE THIS NEUTROPHIL 815 00:28:40,840 --> 00:28:43,840 INFLAMMATION PHENOTYPE AND THIS 816 00:28:43,840 --> 00:28:44,800 AFFORDS THE IDEA THAT MYELOID 817 00:28:44,800 --> 00:28:47,800 GROWTH FACTOR IS WORKING THROUGH 818 00:28:47,800 --> 00:28:50,440 HIP1 ALPHA PATHWAY SO JUST TO 819 00:28:50,440 --> 00:28:51,520 SUMMARIZE MYELOID DRIVE DPROAGHT 820 00:28:51,520 --> 00:28:53,160 FACTOR EFFECTS ON WOUND HEALING. 821 00:28:53,160 --> 00:28:54,560 SO IT'S INDUCED BY STERILE 822 00:28:54,560 --> 00:28:57,960 INJURY BUT NOT INFECTION, IT 823 00:28:57,960 --> 00:28:59,360 LIMITS NEUTROPHIL INFLAMMATION, 824 00:28:59,360 --> 00:29:02,680 AND IT'S A DAMAGE SIGNAL THAT 825 00:29:02,680 --> 00:29:04,960 ACTS VIA HIF1 ALPHA PATHWAY AND 826 00:29:04,960 --> 00:29:07,240 SEEMS TO PROMOTE NEUTROPHIL 827 00:29:07,240 --> 00:29:08,760 REVERSE MIGRATION AND 828 00:29:08,760 --> 00:29:09,440 INFLAMMATION RESOLUTION. 829 00:29:09,440 --> 00:29:12,480 SO A QUESTION IS, HOW DOES IT 830 00:29:12,480 --> 00:29:12,840 WORK? 831 00:29:12,840 --> 00:29:14,560 WHAT IS IT'S RECEPTOR, WHAT'S 832 00:29:14,560 --> 00:29:16,960 THE SOURCES AND TISSUES? 833 00:29:16,960 --> 00:29:18,960 AND ARE THERE--IS THERE A ROLE 834 00:29:18,960 --> 00:29:20,480 FOR MACROPHAGES BECAUSE IN THE 835 00:29:20,480 --> 00:29:22,680 MOUSE STUDIES, IT SEEMS LIKE 836 00:29:22,680 --> 00:29:26,720 MYELOID DRIVE GROWTH FACTOR IN 837 00:29:26,720 --> 00:29:27,320 MACROPHAGES WAS PARTICULARLY 838 00:29:27,320 --> 00:29:29,160 IMPORTANT SO WHAT DO WE KNOW 839 00:29:29,160 --> 00:29:30,600 ABOUT MACROPHAGES IN NEUTROPHILS 840 00:29:30,600 --> 00:29:32,960 AND HOW THEY INTERACT AT SITES 841 00:29:32,960 --> 00:29:33,680 OF TISSUE DAMAGE. 842 00:29:33,680 --> 00:29:39,000 SO IN EARLIER WORK, WE HAD SHOWN 843 00:29:39,000 --> 00:29:41,160 THAT WHEN NEUTROPHILS COME TO 844 00:29:41,160 --> 00:29:43,080 THE WOUND FIRST AND MACROPHAGES 845 00:29:43,080 --> 00:29:44,600 APPEAR, THE NEUTROPHILS LEAVE. 846 00:29:44,600 --> 00:29:47,440 THIS IS THE CASE WITH 847 00:29:47,440 --> 00:29:49,640 MACROPHAGES AND STERILE 848 00:29:49,640 --> 00:29:51,200 INFLAMMATION, NOT IN INFECTED 849 00:29:51,200 --> 00:29:53,520 DAMAGE, SO THESE ARE M2 850 00:29:53,520 --> 00:29:54,800 MACROPHAGES. 851 00:29:54,800 --> 00:29:58,760 AND IF YOU DEPLETE MACROPHAGES, 852 00:29:58,760 --> 00:30:00,600 USING IRF8 MORPHOLIN O, THEN 853 00:30:00,600 --> 00:30:01,360 MORE NEUTROPHILS ARE 854 00:30:01,360 --> 00:30:02,720 PRESCRIBINGENT AT THE WOUND AND 855 00:30:02,720 --> 00:30:03,960 THEY HAVE IMPAIRED REVERSE 856 00:30:03,960 --> 00:30:05,680 MIGRATION SO IT'S 1 OF THE 857 00:30:05,680 --> 00:30:07,520 PATHWAYS THAT'S INVOLVED TBH 858 00:30:07,520 --> 00:30:10,320 THIS NEUTROPHIL REVERSE 859 00:30:10,320 --> 00:30:12,560 MIGRATION AND NICE WORK DONE BY 860 00:30:12,560 --> 00:30:14,640 TD GERMANE LAB IN MOUSE MODELS, 861 00:30:14,640 --> 00:30:16,560 THEY FOUND THAT MACROPHAGES CAN 862 00:30:16,560 --> 00:30:18,720 CLOAK A WOUND AND THAT THIS 863 00:30:18,720 --> 00:30:21,680 INHIB THORS THES NEUTROPHIL 864 00:30:21,680 --> 00:30:22,000 INFLAMMATION. 865 00:30:22,000 --> 00:30:23,920 SO IT SUGGESTS THAT THERE'S SOME 866 00:30:23,920 --> 00:30:27,440 MACROPHAGES ARE VERY INHIBITORY 867 00:30:27,440 --> 00:30:28,760 TO NEUTROPHIL BEHAVIOR. 868 00:30:28,760 --> 00:30:30,000 SO THIS RAISES THE WHOLE 869 00:30:30,000 --> 00:30:32,080 QUESTION LIKE WE ALWAYS TALK 870 00:30:32,080 --> 00:30:34,440 ABOUT M1, M2 MACROPHAGES, WITHIN 871 00:30:34,440 --> 00:30:36,080 TISSUES AND SO, A VERY TALENTED 872 00:30:36,080 --> 00:30:43,160 POST DOC IN THE LAB, VERONICA 873 00:30:43,160 --> 00:30:44,960 MISKOLCI, WAS INTERESTED IN 874 00:30:44,960 --> 00:30:46,800 TRYING TO UNDERSTAND MACROPHAGE 875 00:30:46,800 --> 00:30:47,400 ACTIVATION PHENOTYPES WITHIN 876 00:30:47,400 --> 00:30:52,360 TISSUES AND SHE DID THIS BY 877 00:30:52,360 --> 00:30:54,640 TAKING CARE OF ENDOGENOUS 878 00:30:54,640 --> 00:30:58,240 FLUORESCENCE IN COLLABORATION 879 00:30:58,240 --> 00:30:59,560 WITH MELISSA SKALA, SO CURRENTLY 880 00:30:59,560 --> 00:31:02,160 WHAT WE DO IN ZEBRA FISH TO READ 881 00:31:02,160 --> 00:31:04,960 OUT IN 1 MACROPHAGE IS WE USE A 882 00:31:04,960 --> 00:31:06,280 TNF ALPHA REPORTER. 883 00:31:06,280 --> 00:31:07,560 FOR M2 THERE REALLY HASN'T BEEN 884 00:31:07,560 --> 00:31:12,040 MUCH THAT'S BEEN USED, BUT THE 885 00:31:12,040 --> 00:31:12,760 RGENERATEDAISE REPORTER JUST 886 00:31:12,760 --> 00:31:14,560 CAME OUT BUT I SHOULD SAY WHEN 887 00:31:14,560 --> 00:31:16,560 YOU HAVE THIS TNF ALPHA REPORTER 888 00:31:16,560 --> 00:31:19,000 TURNED ON, THERE TENDS TO BE 889 00:31:19,000 --> 00:31:20,400 MORE PERSISTENT NEUTROPHIL 890 00:31:20,400 --> 00:31:21,560 INFLATION, THOSE DON'T SEEM TO 891 00:31:21,560 --> 00:31:25,360 CARE THE NEUTROPHILS AWAY. 892 00:31:25,360 --> 00:31:30,640 SO WHAT VERONICA USED WAS 1, SO 893 00:31:30,640 --> 00:31:31,880 FLUORESCENCE LIFETIME IMAGING 894 00:31:31,880 --> 00:31:33,680 MICROSCOPY IS A TOOL THAT YOU 895 00:31:33,680 --> 00:31:36,640 CAN USE TO MEASURE INTRA 896 00:31:36,640 --> 00:31:45,960 CELLULAR METABOLISM OF WOUND 897 00:31:45,960 --> 00:31:46,520 MACROPHAGES WITHIN TISSUES. 898 00:31:46,520 --> 00:31:48,000 NOW SLEE DID A LOT OF WORK, THIS 899 00:31:48,000 --> 00:31:50,280 TOOK OVER A YEAR OF REVISIONS, 900 00:31:50,280 --> 00:31:52,560 AND IN PART BECAUSE IT WENT 901 00:31:52,560 --> 00:31:54,560 AGAINST THE DOGMA OF WHAT WE 902 00:31:54,560 --> 00:31:54,880 KNOW INVITRO. 903 00:31:54,880 --> 00:32:08,040 SO WHAT SHE FOUND WAS THAT IN 904 00:32:08,040 --> 00:32:09,040 WOUNDED TISSUES THEY WERE 905 00:32:09,040 --> 00:32:11,160 MACROPHAGE PRESENT, THEY WERE 906 00:32:11,160 --> 00:32:13,640 NOT TNP ALPHA POSITIVE, BUT WITH 907 00:32:13,640 --> 00:32:16,640 WE BURNED THE TISSUE INITIALLY, 908 00:32:16,640 --> 00:32:17,560 WE HADOXIDATIVE TISSUES 909 00:32:17,560 --> 00:32:18,680 RECRUITED AND I AM JUST 910 00:32:18,680 --> 00:32:20,200 SUMMARIZING WHAT I WILL SHOW YOU 911 00:32:20,200 --> 00:32:22,400 AND THESE HAVE BEEN TURNED 912 00:32:22,400 --> 00:32:23,480 NIGGATIVE AS INFLAMMATION 913 00:32:23,480 --> 00:32:23,760 RESOLVES. 914 00:32:23,760 --> 00:32:27,200 SO LET ME SHOW YOU SOME OF THIS 915 00:32:27,200 --> 00:32:27,400 DATA. 916 00:32:27,400 --> 00:32:29,000 SO THE FIRST MODEL I'M SHOWING 917 00:32:29,000 --> 00:32:30,880 YOU IS 1 WHERE WE HAVE INFECTED 918 00:32:30,880 --> 00:32:31,960 A WOUND. 919 00:32:31,960 --> 00:32:34,720 SO THIS TURNS ON TNF ALPHA AND 920 00:32:34,720 --> 00:32:37,760 ALL OF THE MACROPHAGES YOU GET 921 00:32:37,760 --> 00:32:38,600 THIS ROBUST INFLAMMATORY 922 00:32:38,600 --> 00:32:42,160 RESPONSE AND YOU DON'T GET ANY 923 00:32:42,160 --> 00:32:44,560 HEALING AND HERE'S YOU CAN SEE 924 00:32:44,560 --> 00:32:44,880 TNF STAYS ON. 925 00:32:44,880 --> 00:32:47,600 SO WHAT HAPPENS TO THE 926 00:32:47,600 --> 00:32:50,960 METABOLISM OF THESE MACROPHAGES 927 00:32:50,960 --> 00:32:51,680 USING FLEM? 928 00:32:51,680 --> 00:32:54,320 SO WHAT SHE FOUND IS THAT 929 00:32:54,320 --> 00:32:56,360 MACROPHAGES ARE MORE OXIDIZED 930 00:32:56,360 --> 00:32:58,760 WITH THE INCREASED NADPH 931 00:32:58,760 --> 00:33:00,480 LIFETIME AND WE THINK NADPH 932 00:33:00,480 --> 00:33:01,400 LIFETIME IS A PARTICULARLY 933 00:33:01,400 --> 00:33:03,560 USEFUL TOOL TO USE AND A REDUCED 934 00:33:03,560 --> 00:33:05,120 REDOCKS RATIO AT THE INFECTED 935 00:33:05,120 --> 00:33:07,280 WOUND AND YOU CAN SEE THIS HERE. 936 00:33:07,280 --> 00:33:10,200 SO AS I SAID, THIS GOES AGAINST 937 00:33:10,200 --> 00:33:12,960 WHAT WAS--HAD BEEN PUBLISHED 938 00:33:12,960 --> 00:33:13,240 INVITRO. 939 00:33:13,240 --> 00:33:14,760 SO WHAT HAPPENS IN A WOUND WHERE 940 00:33:14,760 --> 00:33:17,720 YOU HAVE THE DYNAMIC CHANGES IN 941 00:33:17,720 --> 00:33:20,360 MACROPHAGES, FIRST 13 ARE 942 00:33:20,360 --> 00:33:21,160 POSITIVE THEN TNF NEGATIVE 943 00:33:21,160 --> 00:33:22,600 BEFORE YOU ACTUALLY GET HEALING 944 00:33:22,600 --> 00:33:25,160 AND YOU CAN SEE HERE, YOU GET 945 00:33:25,160 --> 00:33:27,400 IMPAIRED HEALING BUT EVENTUALLY, 946 00:33:27,400 --> 00:33:28,560 A BURN WOUND DOES HEAL. 947 00:33:28,560 --> 00:33:31,360 WHAT YOU CAN SEE HERE WITH TNF, 948 00:33:31,360 --> 00:33:32,600 ASTERISKS FIRST IT'S ON, THEN 949 00:33:32,600 --> 00:33:33,760 IT'S OFF PRIOR TO HEALING AND 950 00:33:33,760 --> 00:33:37,800 THEN CAN YOU SEE HERE, AGAIN, 951 00:33:37,800 --> 00:33:39,480 NEUTROPHIL INFLAMMATION'S HIGH 952 00:33:39,480 --> 00:33:40,920 WHEN THE MACROPHAGES ARE 13 AT 953 00:33:40,920 --> 00:33:43,160 POSITIVE AND IT RESOLVES AS THEY 954 00:33:43,160 --> 00:33:46,920 TURN TNF NEGATIVE AND THEN THE 955 00:33:46,920 --> 00:33:47,240 TISSUE HEALS. 956 00:33:47,240 --> 00:33:51,240 SO WHEN SHE LOOKED AT THE 957 00:33:51,240 --> 00:33:52,200 METABOLT-ONTIC PROFILE HERE, 958 00:33:52,200 --> 00:33:55,160 MACROPHAGES THAT WERE BURNED 959 00:33:55,160 --> 00:33:56,680 WERE MORE OXIDIZED EARLIER WITH 960 00:33:56,680 --> 00:33:59,560 RESOLUTION BY 72 HOURS SO AGAIN, 961 00:33:59,560 --> 00:34:01,960 TNF ALPHA POSITIVE HIGH 962 00:34:01,960 --> 00:34:03,600 MACROPHAGES HAVE LOW NADPH 963 00:34:03,600 --> 00:34:06,440 LIFETIME AND LOW REDOCKS RATIOS. 964 00:34:06,440 --> 00:34:08,200 AND THIS IS SUMMARIZED HERE, I 965 00:34:08,200 --> 00:34:10,080 GUESS I WENT THROUGH THIS SO I 966 00:34:10,080 --> 00:34:11,960 WON'T GO THROUGH IT IN DETAIL 967 00:34:11,960 --> 00:34:13,600 BUT JUST THIS WHOLE IDEA THAT 968 00:34:13,600 --> 00:34:15,760 THIS IS A DYNAMIC CHANGE OF 969 00:34:15,760 --> 00:34:17,320 ACTIVATION AND TISSUES THAT CAN 970 00:34:17,320 --> 00:34:27,560 BE READ OUT BY LOOKINGA THE 971 00:34:27,560 --> 00:34:28,280 LIFETIMES. 972 00:34:28,280 --> 00:34:30,160 SO CAN WE MODULATE MACROPHAGE 973 00:34:30,160 --> 00:34:32,640 ACTIVATION AND DETECT 974 00:34:32,640 --> 00:34:35,120 CORRESPONDING MET A--BITS OLDER 975 00:34:35,120 --> 00:34:37,560 PEOPLIC CHANGES WITH FLIM, WE 976 00:34:37,560 --> 00:34:40,240 FOUND A REDUCED REDOCKS RATIO 977 00:34:40,240 --> 00:34:42,240 NADPH LIFETIME, MORE NEUTROPHILS 978 00:34:42,240 --> 00:34:44,760 IN THE TISSUES AND ON THE OTHER 979 00:34:44,760 --> 00:34:46,360 EXTREME SHE TREATED WITH MET 980 00:34:46,360 --> 00:34:49,160 FORMIN WHICH LEADS TO N2 981 00:34:49,160 --> 00:34:50,600 MACROPHAGES AND REDUCED POSITIVE 982 00:34:50,600 --> 00:34:51,560 CELLS AND FEWER NEUTROPHILS AND 983 00:34:51,560 --> 00:34:53,160 ELECTRIC TO SEE WHAT LAPPED TO 984 00:34:53,160 --> 00:34:53,560 WOUND HEALING. 985 00:34:53,560 --> 00:34:56,000 AND THAT WAS A METABOLIC SWITCH 986 00:34:56,000 --> 00:34:58,000 IN MACROPHAGES THAT CORRELATES 987 00:34:58,000 --> 00:35:00,600 TO NEUTROPHIL INFLAMMATION AND 988 00:35:00,600 --> 00:35:00,960 WOUND HEALING. 989 00:35:00,960 --> 00:35:03,120 SO WHAT YOU CAN SEE HERE IS WITH 990 00:35:03,120 --> 00:35:06,160 MET FORMIN, THE WOUNDS HEAL 991 00:35:06,160 --> 00:35:09,280 BETTER WITH THE STAT 6 MUTANT, 992 00:35:09,280 --> 00:35:11,000 THE WOUNDS HEALED POORER. 993 00:35:11,000 --> 00:35:12,920 SO THIS SUPPORTS THE IDEA THAT 994 00:35:12,920 --> 00:35:16,920 WE CAN ACTUALLY READ OUT DYNAMIC 995 00:35:16,920 --> 00:35:18,120 CHANGES IN MACROPHAGE PHENOTYPE 996 00:35:18,120 --> 00:35:20,120 BY USING FLIM. 997 00:35:20,120 --> 00:35:23,160 SO HOW DOES MYELOID DRIVE GROWTH 998 00:35:23,160 --> 00:35:24,760 FACTOR EFFECT MACROPHAGE 999 00:35:24,760 --> 00:35:25,760 ACTIVATION? 1000 00:35:25,760 --> 00:35:26,600 THIS IS VERY PRELIMINARY DATA. 1001 00:35:26,600 --> 00:35:29,760 ONE THING WE FOUND IS THAT TNF 1002 00:35:29,760 --> 00:35:32,000 ALPHA IS NOT INCREASED IN 1003 00:35:32,000 --> 00:35:32,800 MACROPHAGES AND MYELOID FACTOR 1004 00:35:32,800 --> 00:35:35,480 MUTANT SO WE THINK THEY'RE MORE 1005 00:35:35,480 --> 00:35:36,760 ACTIVATED BUT WE CAN'T DETECT 1006 00:35:36,760 --> 00:35:39,560 THIS WITH THE TNF ALPHA REPORTER 1007 00:35:39,560 --> 00:35:40,480 BUT IN VERY PRELIMINARY DATA 1008 00:35:40,480 --> 00:35:43,560 WHERE SHE DID THE FLEM ANALYSIS, 1009 00:35:43,560 --> 00:35:45,560 SHE FOUND THAT NADPH LIFETIME 1010 00:35:45,560 --> 00:35:47,200 WAS REDUCED EVEN THOUGH WE 1011 00:35:47,200 --> 00:35:48,960 COULDN'T PICK THIS UP WITH THE 1012 00:35:48,960 --> 00:35:52,120 TNF ALPHA REPORTER AND THAT 1013 00:35:52,120 --> 00:35:54,000 THERE WAS A DECREASE ALSO--SORRY 1014 00:35:54,000 --> 00:35:56,080 THE REDOCKS RATIO WAS REDUCED 1015 00:35:56,080 --> 00:35:58,920 AND THE NADPH LIFETIME, SO THESE 1016 00:35:58,920 --> 00:36:02,280 MACROPHAGES ARE MORE OXIDIZED 1017 00:36:02,280 --> 00:36:03,120 WITH RECREASED LIFETIME. 1018 00:36:03,120 --> 00:36:04,760 SO, WHAT OTHER MARKERS CAN WE 1019 00:36:04,760 --> 00:36:07,960 USE TO TRY TO READ OUT 1020 00:36:07,960 --> 00:36:08,640 MACROPHAGE PHENOTYPES AND WHAT 1021 00:36:08,640 --> 00:36:10,800 DOES THIS MEAN FOR INFLAMMATION? 1022 00:36:10,800 --> 00:36:12,560 SO 1 THATY WOO WERE INTERESTED 1023 00:36:12,560 --> 00:36:15,640 IN WAS IMMUNO RESPONSIVE GENE 1, 1024 00:36:15,640 --> 00:36:22,720 IRG 1 SO IT'S AN ENZYME THAT 1025 00:36:22,720 --> 00:36:24,680 LEADS TO GENERATION, AND 1026 00:36:24,680 --> 00:36:28,200 ITACONNATE AND IT CAN BE 1027 00:36:28,200 --> 00:36:29,280 ANTIMICROBIAL AND INFLAMMATORY 1028 00:36:29,280 --> 00:36:32,720 AND IN ZEBRA FISH CHRIS HALL'S 1029 00:36:32,720 --> 00:36:35,280 LAB REPORTED ZEBRA FISH GFP 1 1030 00:36:35,280 --> 00:36:37,080 REPORTER LINE THAT GOES ON WITH 1031 00:36:37,080 --> 00:36:37,360 INFECTION. 1032 00:36:37,360 --> 00:36:38,640 SO WHAT HAPPENS WITH STERILE 1033 00:36:38,640 --> 00:36:39,200 INJURY? 1034 00:36:39,200 --> 00:36:41,120 SO WHAT HAPPENS WITH STERILE 1035 00:36:41,120 --> 00:36:42,960 INJURY IS WHAT VERONICA FOUND IS 1036 00:36:42,960 --> 00:36:45,720 THAT A SUBSET OF MACROPHAGES 1037 00:36:45,720 --> 00:36:46,920 TURN ON IRG 1. 1038 00:36:46,920 --> 00:36:49,720 SO YOU CAN SEE THAT WITH 1039 00:36:49,720 --> 00:36:51,000 THE--WITH THE MACROPHAGES THAT 1040 00:36:51,000 --> 00:36:52,960 TURN WHITE HERE ARE THE 1041 00:36:52,960 --> 00:36:54,880 MACROPHAGES THAT HAVE TURNED ON 1042 00:36:54,880 --> 00:36:55,280 IRG 1. 1043 00:36:55,280 --> 00:36:58,920 NOT ALL OF THEM, JUST A SUBSET 1044 00:36:58,920 --> 00:36:59,960 OF THEM. 1045 00:36:59,960 --> 00:37:01,480 HOW DOES MYELOID DRIVE GROWTH 1046 00:37:01,480 --> 00:37:02,640 FACTOR AFFECT THIS PHENOTYPE. 1047 00:37:02,640 --> 00:37:05,160 WHEN YOU LOOK AT CONTROL MORPH O 1048 00:37:05,160 --> 00:37:06,400 LINE O VERSUS MYELOID DRIVE 1049 00:37:06,400 --> 00:37:07,000 GROWTH FACTOR, WHAT YOU SLEEP 1050 00:37:07,000 --> 00:37:09,480 APNEA AND OBESITYY IS THESIS 1051 00:37:09,480 --> 00:37:11,400 MACROPHAGES TURN ON IRG 1, SO 1052 00:37:11,400 --> 00:37:15,920 THERE'S MUCH MORE IRG 1 REPORTER 1053 00:37:15,920 --> 00:37:17,680 ACTIVITY WHEN YOU DEPLOAT 1054 00:37:17,680 --> 00:37:18,400 MYELOID DRIVE GROWTH FACTOR SO 1055 00:37:18,400 --> 00:37:21,160 THIS IS A DIFFERENT TYPE OF 1056 00:37:21,160 --> 00:37:23,120 ACTIVATED MACK RAPHAGE. 1057 00:37:23,120 --> 00:37:24,960 SO JUST--MACROPHAGE SO JUST AS A 1058 00:37:24,960 --> 00:37:27,080 HUMBLING WAY OF SAYING THESE 1059 00:37:27,080 --> 00:37:29,600 THINGS ARE VERY COMPLICATED, 1060 00:37:29,600 --> 00:37:32,000 MYELOID GROWTH FACTOR ACTIVATES 1061 00:37:32,000 --> 00:37:33,440 AND DRIVES METABOLISM SO ARE 1062 00:37:33,440 --> 00:37:34,880 THERE NEUTROPHIL CHANGES DUE TO 1063 00:37:34,880 --> 00:37:37,760 THE CHANGES IN MACROPHAGES OR IS 1064 00:37:37,760 --> 00:37:39,560 THERE SOME NEUTROPHIL INTRINSIC 1065 00:37:39,560 --> 00:37:40,600 ASPECT OF THIS AND WE DON'T KNOW 1066 00:37:40,600 --> 00:37:44,080 THE ANSWER TO THAT. 1067 00:37:44,080 --> 00:37:47,280 SO--WELL I GIVE A BIT OF DATA I 1068 00:37:47,280 --> 00:37:48,400 THINK IS INTRIGUING. 1069 00:37:48,400 --> 00:37:50,240 SO MACK RAPHAGE ACTIVATION 1070 00:37:50,240 --> 00:37:52,080 ASSOCIATE WIDE METABOLIC 1071 00:37:52,080 --> 00:37:53,000 REPROGRAMMING INVIVO, WE ARE 1072 00:37:53,000 --> 00:37:54,160 INTERESTED IN THE CROSS TALK 1073 00:37:54,160 --> 00:37:55,960 BETWEEN THESE DIFFERENT TYPES OF 1074 00:37:55,960 --> 00:37:57,040 MACROPHAGES AND NEUTROPHILS AND 1075 00:37:57,040 --> 00:37:59,160 HOW IT PLAYS A ROLE IN 1076 00:37:59,160 --> 00:38:00,760 INFLAMMATION, SO HOW CAN WE 1077 00:38:00,760 --> 00:38:04,200 FURTHER GO AFTER THIS PATHWAY. 1078 00:38:04,200 --> 00:38:06,160 WELL, 1 WAY IS TO LOOK AT 1079 00:38:06,160 --> 00:38:08,920 DIFFERENT TYPES OF TISSUE DAMAGE 1080 00:38:08,920 --> 00:38:14,720 AND THIS IS IN THE CASE OF 1081 00:38:14,720 --> 00:38:14,960 MELANOMA. 1082 00:38:14,960 --> 00:38:16,520 RICHARD WHITE'S LAB SOME 1083 00:38:16,520 --> 00:38:17,640 BIOARCHIVES PAPER AND WE 1084 00:38:17,640 --> 00:38:19,000 COLLABORATED WITH THEM TO LOOK 1085 00:38:19,000 --> 00:38:21,280 AT WHAT HAPPENS TO MYELOID 1086 00:38:21,280 --> 00:38:22,680 DRIIVE GROWTH FACTOR IN THE 1087 00:38:22,680 --> 00:38:25,320 TUMOR MICROENVIRONMENT AND YOU 1088 00:38:25,320 --> 00:38:28,120 CAN SEE THAT MYELOID DERIVED 1089 00:38:28,120 --> 00:38:32,160 GROWTH FACTOR IS DRAMATICALLY 1090 00:38:32,160 --> 00:38:33,880 INCREASED. 1091 00:38:33,880 --> 00:38:35,440 SO GAYATHRI, YOU A POST DOC IN 1092 00:38:35,440 --> 00:38:36,880 THE LAB AND TRYING FIGURE OUT 1093 00:38:36,880 --> 00:38:38,480 WHAT THIS MEANS AND THE ISHT 1094 00:38:38,480 --> 00:38:40,160 ACTION BETWEEN NEUTROPHILS AND 1095 00:38:40,160 --> 00:38:40,560 MACROPHAGES. 1096 00:38:40,560 --> 00:38:44,200 TO ALSO GO AFTER THIS QUESTION, 1097 00:38:44,200 --> 00:38:45,840 WE HAVE GENERATED HIMAVEN IPS 1098 00:38:45,840 --> 00:38:48,880 CELLS THAT ARE DEFICIENT IN 1099 00:38:48,880 --> 00:38:51,480 MYELOID DERIVED GROWTH FACTOR 1100 00:38:51,480 --> 00:38:57,120 AND IN COLLABORATION WITH 1101 00:38:57,120 --> 00:38:57,760 IGOR WE HAVE GENERATED HEPATITIS 1102 00:38:57,760 --> 00:39:00,560 EAT O FILLS THAT ARE INTERESTING 1103 00:39:00,560 --> 00:39:01,720 FOR PURPOSES SO GENERATING 1104 00:39:01,720 --> 00:39:02,320 NEUTROPHILS THAT MIGHT HAVE 1105 00:39:02,320 --> 00:39:05,120 FUNCTIONS THAT WILL BE USEFUL 1106 00:39:05,120 --> 00:39:08,200 LIKE ENGINEERING, NEUTROPHILS TO 1107 00:39:08,200 --> 00:39:09,160 TREAT VARIOUS HUMAN DISEASES, 1108 00:39:09,160 --> 00:39:11,160 AND SHIP THEM TOWARDS AN 1109 00:39:11,160 --> 00:39:16,640 ANTITUMOR PHENOTYPE FOR EXAMPLE, 1110 00:39:16,640 --> 00:39:20,040 CAR NEUTROPHILS VERSUS A PRO 1111 00:39:20,040 --> 00:39:20,400 TUMOR PHENOTYPE. 1112 00:39:20,400 --> 00:39:22,560 AND THEN GOING FORWARD, TO TRY 1113 00:39:22,560 --> 00:39:26,200 TO GET MORE AT THIS PATHWAY, IN 1114 00:39:26,200 --> 00:39:27,320 ADDITION TO UNDERSTANDING OTHER 1115 00:39:27,320 --> 00:39:30,560 MECHANISMS THAT ARE INVOLVED IN 1116 00:39:30,560 --> 00:39:31,480 NEUTROPHIL BEHAVIOR AND 1117 00:39:31,480 --> 00:39:33,920 MACROPAGE BEHAVIOR AND TISSUE, 1118 00:39:33,920 --> 00:39:36,160 WERE INTERESTED IN UNDERSTANDING 1119 00:39:36,160 --> 00:39:38,800 THE DIVERSITY OF MACROPHAGES AND 1120 00:39:38,800 --> 00:39:40,600 INTERSTITIAL TISSUES USING LIECH 1121 00:39:40,600 --> 00:39:42,000 IMAGES AND MORE SINGLE CELL 1122 00:39:42,000 --> 00:39:44,200 ANALYSIS AND AS WELL AS SINGLE 1123 00:39:44,200 --> 00:39:45,160 CELL TRANSCRIPT OHMICS AND THIS 1124 00:39:45,160 --> 00:39:46,960 IS DONE IN COLLABORATION WITH 1125 00:39:46,960 --> 00:39:48,880 [INDISCERNIBLE] AN ASSIST ABT 1126 00:39:48,880 --> 00:39:50,600 PROFESSOR AT UW MADISON. 1127 00:39:50,600 --> 00:39:53,360 AND SO, WHEN YOU LOOK AT THE 1128 00:39:53,360 --> 00:39:55,080 BEHAVIOR OF NEUTROPHILS AND 1129 00:39:55,080 --> 00:39:57,320 MACROPHAGES AND RESPONSE TO A 1130 00:39:57,320 --> 00:39:57,760 BURN WOUND. 1131 00:39:57,760 --> 00:40:01,280 YOU CAN SEE THAT THE MACROPHAGES 1132 00:40:01,280 --> 00:40:01,840 AND NEUTROPHILS BEHAVIOR 1133 00:40:01,840 --> 00:40:02,560 DIFFERENTLY, MACROPHAGES DON'T 1134 00:40:02,560 --> 00:40:05,960 LIKE TO GO INTO THE BURN TISSUE 1135 00:40:05,960 --> 00:40:07,160 WHEREAS NEUTROPHILS MIGRATE VERY 1136 00:40:07,160 --> 00:40:08,480 RAPIDLY WITHIN THIS BURN TISSUE. 1137 00:40:08,480 --> 00:40:11,040 SO WHAT'S GOING ON HERE, WHEN 1138 00:40:11,040 --> 00:40:13,880 YOU BURN THE TAIL OF A ZEBRA 1139 00:40:13,880 --> 00:40:15,600 FISH AND CONTROL, WHEN YOU CUT 1140 00:40:15,600 --> 00:40:19,640 OFF THE TAIL, YOU SEE THIS NICE 1141 00:40:19,640 --> 00:40:20,760 COLLAGEN ORGANIZATION BY SECOND 1142 00:40:20,760 --> 00:40:22,440 HARMONIC IMAGING BUT WHEN YOU 1143 00:40:22,440 --> 00:40:25,200 BURN THE TAIL, YOU LOSE THIS 1144 00:40:25,200 --> 00:40:25,640 ORGANIZED COLLAGEN. 1145 00:40:25,640 --> 00:40:29,280 SO AS I MENTIONED TO YOU 1146 00:40:29,280 --> 00:40:31,080 EARLIER, NEUTROPHILS ARE CLASSIC 1147 00:40:31,080 --> 00:40:33,560 AMOEBOID CELLS THEY MIGRATE 1148 00:40:33,560 --> 00:40:35,640 INDEPENDENT OF INDEPENDENT 1149 00:40:35,640 --> 00:40:38,640 MEDIATED HESION AND AND MIGRATE 1150 00:40:38,640 --> 00:40:40,000 IN THIS COLLAGEN FREE ZONE WHERE 1151 00:40:40,000 --> 00:40:42,720 THESE ARE MORE MEASEN CHIMAL AND 1152 00:40:42,720 --> 00:40:43,480 MIGRATION IS SIGNIFICANTLY 1153 00:40:43,480 --> 00:40:47,280 IMPAIRED SO THIS IS THE SORT OF 1154 00:40:47,280 --> 00:40:50,080 SINGLE CELL BEHAVIOR, GROUP 1155 00:40:50,080 --> 00:40:52,240 BEHAVIOR THAT WE'RE DOING MORE 1156 00:40:52,240 --> 00:40:54,240 OF, BUT IN ADDITION IN THIS SAME 1157 00:40:54,240 --> 00:40:57,640 BURN MODEL WHAT WE'VE DONE S&P A 1158 00:40:57,640 --> 00:41:00,400 SINGLE CELL RNA SEQ ANALYSIS OF 1159 00:41:00,400 --> 00:41:01,760 NEUTROPHILS AND MACROPHAGES AND 1160 00:41:01,760 --> 00:41:03,360 THIS IS VERY PRELIMINARY NEW 1161 00:41:03,360 --> 00:41:04,840 DATA THAT'S COME OUT BUT IT'S 1162 00:41:04,840 --> 00:41:06,520 VERY INTERESTING THAT WHEN YOU 1163 00:41:06,520 --> 00:41:10,960 LOOK 24 HOURS AFTER BURN, YOU 1164 00:41:10,960 --> 00:41:13,360 CAN SEE THAT THEIR DISTINCT 1165 00:41:13,360 --> 00:41:16,560 POPULATIONS OF NEUTROPHILS, AND 1166 00:41:16,560 --> 00:41:18,320 DISTINCT POPULATIONS OF 1167 00:41:18,320 --> 00:41:19,240 MACROPHAGES THAT ARE SEPARABLE, 1168 00:41:19,240 --> 00:41:20,520 WHAT WAS ALSO INTERESTING IS 1169 00:41:20,520 --> 00:41:22,040 THERE'S SHIFT IN THE POPULATIONS 1170 00:41:22,040 --> 00:41:24,120 WITH A BURN WOUND AND IN 1171 00:41:24,120 --> 00:41:25,160 PARTICULAR ALL THE NEUTROPHIL 1172 00:41:25,160 --> 00:41:27,560 SUBSETS WERE INCREASED WITH THE 1173 00:41:27,560 --> 00:41:28,960 BURN WOUND BUT THE MACROPHAGE 1174 00:41:28,960 --> 00:41:30,120 SUBSETS MANY OF THEM WERE 1175 00:41:30,120 --> 00:41:31,680 DECREASED, NOT ALL OF THEM. 1176 00:41:31,680 --> 00:41:34,400 SO CAN WE USE THIS TO TRY TO 1177 00:41:34,400 --> 00:41:38,040 UNDERSTAND MORE OF WHAT'S GOING 1178 00:41:38,040 --> 00:41:38,920 ON WITH NEUTROPHIL BEHAVIOR 1179 00:41:38,920 --> 00:41:42,160 WITHIN TISSUES AND SO, AN 1180 00:41:42,160 --> 00:41:45,280 EXAMPLE OF LOOKING AT A RECEPTOR 1181 00:41:45,280 --> 00:41:49,240 LIGAND INTERACTION IS IL6 AND 1182 00:41:49,240 --> 00:41:49,560 IL6 RECEPTOR. 1183 00:41:49,560 --> 00:41:53,680 SO, IN CONTROL WHAT YOU CAN SEE 1184 00:41:53,680 --> 00:41:56,760 IS THAT THE MACROPHAGES EXPRESS 1185 00:41:56,760 --> 00:42:01,640 IL6 AND THE NEUTROPHILS EXPRESS 1186 00:42:01,640 --> 00:42:05,200 THE IL6 RECEPTOR. 1187 00:42:05,200 --> 00:42:07,560 SO, SO--SORRY HERE'S THE 1188 00:42:07,560 --> 00:42:08,560 NEUTROPHILS EXPRESS THE IL6 1189 00:42:08,560 --> 00:42:10,480 RECEPTOR SO THE LIGANDS BEING 1190 00:42:10,480 --> 00:42:12,040 PRODUCED BY MACROPHAGES AND THEN 1191 00:42:12,040 --> 00:42:13,160 THE NEUTROPHILS EXPRESS THE 1192 00:42:13,160 --> 00:42:16,840 RECEPTOR AND WITH BURN, THE 1193 00:42:16,840 --> 00:42:18,400 RECEPTOR IS SIGNIFICANTLY 1194 00:42:18,400 --> 00:42:19,560 UPREGULATE IN THE NEUTROPHIL 1195 00:42:19,560 --> 00:42:22,160 POPULATION AND THERE'S SOME DOWN 1196 00:42:22,160 --> 00:42:23,760 REGULATION OF IL6, IN THE 1197 00:42:23,760 --> 00:42:25,720 MACROPHAGES, 24 HOURS AFTER A 1198 00:42:25,720 --> 00:42:26,640 BURN. 1199 00:42:26,640 --> 00:42:30,560 SO WHAT DOES IL6 RECEPTOR 1200 00:42:30,560 --> 00:42:31,720 DEPLETION DO TO NEUTROPHIL 1201 00:42:31,720 --> 00:42:32,880 RESPONSES TO A BURN WOUND? 1202 00:42:32,880 --> 00:42:36,240 SO YOU MIGHT PREDICT THAT THE 1203 00:42:36,240 --> 00:42:37,120 NEUTROPHIL MACROPHAGE PENE O 1204 00:42:37,120 --> 00:42:38,560 TYPE WOULD BE VERY DIFFERENT 1205 00:42:38,560 --> 00:42:41,440 BECAUSE THE MACROPHAGES DO NOT 1206 00:42:41,440 --> 00:42:44,600 EXPRESS THE IL6 RECEPTOR. 1207 00:42:44,600 --> 00:42:46,560 INDEED THIS IS WHAT WE FOUND. 1208 00:42:46,560 --> 00:42:49,240 SO IN CONTROL, YOU SEE NICE 1209 00:42:49,240 --> 00:42:51,040 RECRUITMENT OF BOTH NEUTROPHILS 1210 00:42:51,040 --> 00:42:51,640 AND MACROPHAGES. 1211 00:42:51,640 --> 00:42:54,200 WHEN WE USE AN IL6 RECEPTOR 1212 00:42:54,200 --> 00:42:57,360 MUTANT, YOU CAN SEE THAT 1213 00:42:57,360 --> 00:42:59,600 NEUTROPHILS ARE NO LONGER 1214 00:42:59,600 --> 00:43:01,400 RECRUITED TO THE WOUND SITE 1215 00:43:01,400 --> 00:43:04,360 ALTHOUGH MACROPHAGES ARE 1216 00:43:04,360 --> 00:43:04,800 RECRUITED. 1217 00:43:04,800 --> 00:43:06,040 AND THIS IS QUANTIFIED EAR WHERE 1218 00:43:06,040 --> 00:43:08,320 YOU CAN SEE THAT NEUTROPHILS AND 1219 00:43:08,320 --> 00:43:11,160 MACROPHAGES ARE BOTH RECRUITED. 1220 00:43:11,160 --> 00:43:13,800 WHEREAS IN IL6 RECEPTOR MUTANT, 1221 00:43:13,800 --> 00:43:15,160 NEUTROPHIL RECRUITMENT IS 1222 00:43:15,160 --> 00:43:16,360 SIGNIFICANTLY IMPAIRED. 1223 00:43:16,360 --> 00:43:18,960 SO THIS JUST SUPPORTS THE POWER 1224 00:43:18,960 --> 00:43:21,040 OF THE SYSTEM TO TRY TO START 1225 00:43:21,040 --> 00:43:23,960 UNDERSTANDING SOME OF THESE 1226 00:43:23,960 --> 00:43:25,320 RECEPTOR LIGAND INTERACTIONS. 1227 00:43:25,320 --> 00:43:26,840 SO WHAT HAPPENS WITH MYELOID 1228 00:43:26,840 --> 00:43:28,000 DRIVE GROWTH FACTOR AND AGAIN 1229 00:43:28,000 --> 00:43:32,960 THIS IS DATA WE JUST PULLED OFF 1230 00:43:32,960 --> 00:43:35,600 SO IT'S VERY PRELIMINARY 1 OF 1231 00:43:35,600 --> 00:43:37,120 THE PUTATIVE RECEPTORS WOULD BE 1232 00:43:37,120 --> 00:43:38,040 THEICATE O RECEPTORS AND WE 1233 00:43:38,040 --> 00:43:40,880 LOOKEDDA THE ALL THEICATE O 1234 00:43:40,880 --> 00:43:43,320 RECEPTORS VERY INTERESTINGLY WAS 1235 00:43:43,320 --> 00:43:45,200 UP REGULATED IN THE SAME 1236 00:43:45,200 --> 00:43:47,240 NEUTROPHIL POPULATIONS THAT 1237 00:43:47,240 --> 00:43:55,960 UPREGULATED MYELOID DRIVE GROWTH 1238 00:43:55,960 --> 00:43:56,560 FACTOR WITH BURN. 1239 00:43:56,560 --> 00:43:57,360 SO IT'S INTERESTING TO THINK 1240 00:43:57,360 --> 00:44:00,160 ABOUT WHAT THIS MEANS? 1241 00:44:00,160 --> 00:44:03,240 SO DOES IT MAKE THIS RECEPTOR 1242 00:44:03,240 --> 00:44:06,520 AND SIGNAL TO THE RECEPTOR IN AN 1243 00:44:06,520 --> 00:44:09,440 AUTOKRIEN SIGNALING PATHWAY AND 1244 00:44:09,440 --> 00:44:11,240 IN MACROPHAGES ONLY 1 EXPRESSED 1245 00:44:11,240 --> 00:44:13,040 GROWTH FACTOR, THEY ALSO 1246 00:44:13,040 --> 00:44:20,560 EXPRESSED IRG 1 AND THEN AND 1247 00:44:20,560 --> 00:44:24,080 THEN WITH A BURN WOUND THEY ALSO 1248 00:44:24,080 --> 00:44:25,080 EXPRESSED THE KADEL RECEPTOR BUT 1249 00:44:25,080 --> 00:44:26,600 IT WAS NOT CHANGED WITH BURNS. 1250 00:44:26,600 --> 00:44:28,360 SO THESE ARE THE QUESTIONS WE 1251 00:44:28,360 --> 00:44:29,000 ARE GOING AFTER NOW. 1252 00:44:29,000 --> 00:44:31,560 WE CAN USE BOTH THE INVITRO 1253 00:44:31,560 --> 00:44:33,720 SYSTEM AND THE ZEBRA FISH SYSTEM 1254 00:44:33,720 --> 00:44:37,560 TO UNDERSTAND THESE 1255 00:44:37,560 --> 00:44:37,920 INTERACTIONS. 1256 00:44:37,920 --> 00:44:39,760 SO IT'S MORE OF A QUESTION OF 1257 00:44:39,760 --> 00:44:41,720 FUTURE RESEARCH TO UNCOVER 1258 00:44:41,720 --> 00:44:47,160 WHAT'S GOING ON HERE. 1259 00:44:47,160 --> 00:44:48,640 SO JUST TO END I WOULD LIKE TO 1260 00:44:48,640 --> 00:44:51,440 SAY THAT WE'RE VERY INTERESTED 1261 00:44:51,440 --> 00:44:55,360 IN THE BALANCE BETWEEN 1262 00:44:55,360 --> 00:44:57,040 NEUTROPHIL INFLAMMATION AND 1263 00:44:57,040 --> 00:44:58,760 REPAIR, THE FACTORS LIKE MYELOID 1264 00:44:58,760 --> 00:45:00,040 DERIVED GROWTH FACTOR THAT 1265 00:45:00,040 --> 00:45:05,600 IMPACT WHAT'S GOING ON AT 1266 00:45:05,600 --> 00:45:06,480 STERILE INJURY SITES TISSUE 1267 00:45:06,480 --> 00:45:08,240 DAMAGE, BUT NOT IN THE CASE OF 1268 00:45:08,240 --> 00:45:14,360 PATHOGEN OR INFECTED TISSUE. 1269 00:45:14,360 --> 00:45:17,760 AND THIS MAY BE A USEFUL TOOL TO 1270 00:45:17,760 --> 00:45:18,600 LIMIT INFLAMMATION, BUT WE STILL 1271 00:45:18,600 --> 00:45:20,560 DON'T KNOW WHETHER THIS IS 1272 00:45:20,560 --> 00:45:21,480 SECONDAR TOW THE MACROPHAGES 1273 00:45:21,480 --> 00:45:24,160 THAT ARE MAKING IT OR IS IT IN 1274 00:45:24,160 --> 00:45:26,160 FACT, SOME OF IT INTRINSIC TO 1275 00:45:26,160 --> 00:45:27,520 THE NEUTROPHIL, SO THESE ARE THE 1276 00:45:27,520 --> 00:45:33,320 SORTS OF QUESTIONS WE'RE 1277 00:45:33,320 --> 00:45:34,680 INTERESTED IN GOING AFTER SO I 1278 00:45:34,680 --> 00:45:36,440 WOULD LIKE TO JUST END AND 1279 00:45:36,440 --> 00:45:37,080 ACKNOWLEDGE MY RESEARCH GROUP 1280 00:45:37,080 --> 00:45:39,640 AND I WORK WITH A FANTASTIC 1281 00:45:39,640 --> 00:45:41,720 GROUP OF PEOPLE, MY LONG-TERM 1282 00:45:41,720 --> 00:45:43,440 MEMBERS, DAVID AND JULIE, 1283 00:45:43,440 --> 00:45:45,240 DAVID'S BEEN IN THE LAB FOR OVER 1284 00:45:45,240 --> 00:45:47,200 20 YEARS AND JULIE FOR OVER 12 1285 00:45:47,200 --> 00:45:48,920 YEARS HAVE BEEN AMAZING TO WORK 1286 00:45:48,920 --> 00:45:54,080 WITH, AND THEN THE WHOLE TEAM OF 1287 00:45:54,080 --> 00:45:55,840 GRADUATE STUDENTS AND POST DOCS, 1288 00:45:55,840 --> 00:45:58,200 MANY OF WHOM I ACKNOWLEDGED 1289 00:45:58,200 --> 00:46:00,560 DURING THE TALK AND HERE ARE 1290 00:46:00,560 --> 00:46:02,600 FORMER POST DOCS WHO NOW HAVE 1291 00:46:02,600 --> 00:46:05,640 THEIR OWN RESEARCH GROUPS. 1292 00:46:05,640 --> 00:46:07,640 AND COLLABORATORS WHO I 1293 00:46:07,640 --> 00:46:08,520 ACKNOWLEDGED DURING THE TALK. 1294 00:46:08,520 --> 00:46:12,200 I'D LIKE TO END THERE AND ASK IF 1295 00:46:12,200 --> 00:46:22,160 THERE ARE ANY QUESTIONS. 1296 00:46:22,160 --> 00:46:23,560 SO WHICH NADPHOX INHIBITOR 1297 00:46:23,560 --> 00:46:27,200 DID YOU USE IN THE ZEBRA FISH 1298 00:46:27,200 --> 00:46:30,400 WOUND/INFECTION MODEL THAT COMES 1299 00:46:30,400 --> 00:46:30,840 FROM AMELIA? 1300 00:46:30,840 --> 00:46:34,920 >> WE USED DPI SO THIS IS A VERY 1301 00:46:34,920 --> 00:46:37,680 COMMON IN NADPHOX DACE 1302 00:46:37,680 --> 00:46:38,880 INHIBITOR. 1303 00:46:38,880 --> 00:46:41,080 IF YOU DEPLETE P22 FOX YOU COULD 1304 00:46:41,080 --> 00:46:45,440 END UP GETTING MORE NEUTROPHIL 1305 00:46:45,440 --> 00:46:48,560 INFLAMMATION, AND SO, IT'S--IT'S 1306 00:46:48,560 --> 00:46:49,800 COMPLICATED HOW--WHAT THE SOURCE 1307 00:46:49,800 --> 00:46:55,800 IS OF THE NADPH AND THE IMPACT 1308 00:46:55,800 --> 00:46:56,200 ON INFLAMMATION. 1309 00:46:56,200 --> 00:46:57,880 >> THIS WAS REALLY WONDERFUL, I 1310 00:46:57,880 --> 00:47:00,680 HAVE 1 QUESTION REGARDS TO THE 1311 00:47:00,680 --> 00:47:02,040 MILEIN DERIVED GROWTH FACTOR. 1312 00:47:02,040 --> 00:47:04,600 YOU SHOWED THIS IS IMPORTANT FOR 1313 00:47:04,600 --> 00:47:06,240 HEALING OF THE BURN WOUNDS YOU 1314 00:47:06,240 --> 00:47:07,320 DID MENTION BRIEFLY THAT IT IS 1315 00:47:07,320 --> 00:47:09,880 NOT AS IMPORTANT FOR THE 1316 00:47:09,880 --> 00:47:10,280 INFECTIOUS WOUNDS. 1317 00:47:10,280 --> 00:47:11,880 HOW DOES IT ACTUALLY--WHAT ROLE 1318 00:47:11,880 --> 00:47:14,520 DOES IT PLAY OR HAVE YOU LOOKED 1319 00:47:14,520 --> 00:47:16,600 AT ITS ROLE OF OTHER STERILE 1320 00:47:16,600 --> 00:47:18,400 FORMS OF INFLAMMATION THAT ARE 1321 00:47:18,400 --> 00:47:19,360 NOT WOUNDS? 1322 00:47:19,360 --> 00:47:20,440 >> WITH REG TAIL TRANSSECTION, 1323 00:47:20,440 --> 00:47:22,240 IT'S ALSO IMPORTANT FOR HEALING. 1324 00:47:22,240 --> 00:47:24,400 >> SO IT'S THE CUTTING AND THE 1325 00:47:24,400 --> 00:47:24,600 BURN. 1326 00:47:24,600 --> 00:47:26,200 >> THE CUT AND THE BURN. 1327 00:47:26,200 --> 00:47:28,800 WE HAVEN'T LOOKEDDA THE OTHER 1328 00:47:28,800 --> 00:47:31,400 DAMAGE MODELS YET. 1329 00:47:31,400 --> 00:47:33,640 >> OR INFLAMMATORY STERILE 1330 00:47:33,640 --> 00:47:34,000 INFLAMMATION,. 1331 00:47:34,000 --> 00:47:35,080 >> YEAH WE HAVEN'T LOOKED AT 1332 00:47:35,080 --> 00:47:36,400 OTHER MODELS BUT I'M PREETY 1333 00:47:36,400 --> 00:47:39,000 EXCITED ABOUT IT AS A POTENTIAL 1334 00:47:39,000 --> 00:47:40,520 TARGET AND THERE'S OBVIOUSLY 1335 00:47:40,520 --> 00:47:42,560 GROWING INTEREST IN IT FROM ITS 1336 00:47:42,560 --> 00:47:45,080 IMPACT ON MY O CARDIAL 1337 00:47:45,080 --> 00:47:45,480 INFARCTION. 1338 00:47:45,480 --> 00:47:48,400 >> SO ANNA, THAT WAS REALLY 1339 00:47:48,400 --> 00:47:48,640 TERRIFIC. 1340 00:47:48,640 --> 00:47:52,320 YOU TALKED A LITTLE BIT ABOUT 1341 00:47:52,320 --> 00:47:55,560 THE ROLE OF VARIOUS CYTOKINES, 1342 00:47:55,560 --> 00:47:56,400 TNF AND DENDRITIC CELL ROUGH 1343 00:47:56,400 --> 00:48:00,240 ATOM LUKEIN 6 IN SOME OF THESE 1344 00:48:00,240 --> 00:48:01,720 PROCESSES, NEUTROPHIL AND 1345 00:48:01,720 --> 00:48:03,200 MACROPHAGE MEDIATED WOUND 1346 00:48:03,200 --> 00:48:05,800 HEALING FOR EXAMPLE, AND WE USE 1347 00:48:05,800 --> 00:48:08,800 NOW, CLINICALLY, A LOT OF 1348 00:48:08,800 --> 00:48:10,360 INHIBITORS OF TNF AND IL6 AND 1349 00:48:10,360 --> 00:48:13,360 OTHER CYTOKINES AND SO I JUST 1350 00:48:13,360 --> 00:48:15,600 WONDERED IF YOU HAD GIVEN ANY 1351 00:48:15,600 --> 00:48:17,160 THOUGHT TO OR LOOKED AT THE ROLE 1352 00:48:17,160 --> 00:48:20,200 OF SOME OF THESE INHIBITORS WE 1353 00:48:20,200 --> 00:48:21,480 USE CLINICALLY, AND THINKING 1354 00:48:21,480 --> 00:48:22,720 ABOUT WHAT EFFECT THEY MIGHT 1355 00:48:22,720 --> 00:48:26,200 HAVE ON WOUND HEALING IN 1356 00:48:26,200 --> 00:48:26,480 PATIENTS. 1357 00:48:26,480 --> 00:48:28,920 >> YEAH, IF IT'S A TRICKY THING 1358 00:48:28,920 --> 00:48:29,800 WITH ZEBRA FISH BECAUSE 1359 00:48:29,800 --> 00:48:32,040 SOMETIMES YOU DON'T NECESSARILY 1360 00:48:32,040 --> 00:48:33,360 GET CROSS REACTIVITY OF THESE 1361 00:48:33,360 --> 00:48:35,440 ANTIBODIES, BUT THE ZEBRA FISH 1362 00:48:35,440 --> 00:48:36,800 FORM OF THE CYTOKINES. 1363 00:48:36,800 --> 00:48:40,280 THE INTERESTING THING WITH IL6, 1364 00:48:40,280 --> 00:48:42,040 INHIBITION IS IT TOTALLY 1365 00:48:42,040 --> 00:48:44,320 INHIBITS NEUTROPHILS BUT IT DID 1366 00:48:44,320 --> 00:48:47,840 NOT INHIBIT WOUND HEALING WHICH 1367 00:48:47,840 --> 00:48:49,320 IS INTERESTING. 1368 00:48:49,320 --> 00:48:51,640 IN THE CASE OF TNF, IT IS ALSO 1369 00:48:51,640 --> 00:48:52,600 COMPLICATED, THERE'S A LOT OF 1370 00:48:52,600 --> 00:48:55,440 ISSUES WITH TOO MUCH OR TOO 1371 00:48:55,440 --> 00:48:57,800 LITTLE TNF SO IT'S NOT SUCH AS 1372 00:48:57,800 --> 00:49:02,320 EASY EXAMPLE OF SAYING, YOU KNOW 1373 00:49:02,320 --> 00:49:03,560 INHIBITING TNF IS NECESSARILY 1374 00:49:03,560 --> 00:49:04,400 BAD FOR WOUND HEALING. 1375 00:49:04,400 --> 00:49:05,800 SO THIS IS THE THING THAT'S 1376 00:49:05,800 --> 00:49:07,880 REALLY HARD IS THAT THERE'S 1377 00:49:07,880 --> 00:49:10,040 THESE SPECIFIC CELL TYPES THAT 1378 00:49:10,040 --> 00:49:11,760 MAKE THESE PRODUCTS BUT THEY CAN 1379 00:49:11,760 --> 00:49:14,200 FLIP EITHER WAY IN TERMS OF 1380 00:49:14,200 --> 00:49:16,840 THEIR IMPORTANCE WHICH MAKE ITS 1381 00:49:16,840 --> 00:49:17,200 SO CHALLENGING. 1382 00:49:17,200 --> 00:49:20,200 NOW WE KNOW TNF INHIBITION IS 1383 00:49:20,200 --> 00:49:21,280 INCREDIBLY EFFECTIVE FOR 1384 00:49:21,280 --> 00:49:23,160 TREATING OUR PATIENTS, AND IT 1385 00:49:23,160 --> 00:49:26,400 DOESN'T HAVE MANY SIDE EFFECTS, 1386 00:49:26,400 --> 00:49:26,600 RIGHT? 1387 00:49:26,600 --> 00:49:27,200 >> YEAH, YEAH. 1388 00:49:27,200 --> 00:49:29,480 >> ALL RIGHT, WELL, ANYWAY, IT'S 1389 00:49:29,480 --> 00:49:32,600 REALLY INTERESTING. 1390 00:49:32,600 --> 00:49:33,920 >> THANKS. 1391 00:49:33,920 --> 00:49:35,240 >> FASCINATING YES. 1392 00:49:35,240 --> 00:49:42,920 >> ANY OTHER QUESTIONS? 1393 00:49:42,920 --> 00:49:44,960 >> I HAD A QUESTION. 1394 00:49:44,960 --> 00:49:46,920 >> WOBDERFUL TALK BY THE WAY, 1395 00:49:46,920 --> 00:49:49,120 HICURIOSITY RELATED TO WHAT YOU 1396 00:49:49,120 --> 00:49:51,520 MENTIONED ABOUT MACROPHAGES 1397 00:49:51,520 --> 00:49:53,000 VERSUS NEUTROPHIL AND THE 1398 00:49:53,000 --> 00:49:54,720 COLLAGEN AND THE ISSUE, HOW IS 1399 00:49:54,720 --> 00:49:57,040 IT DIFFERENT WHEN YOU HAVE 1400 00:49:57,040 --> 00:49:57,320 INFECTION? 1401 00:49:57,320 --> 00:49:58,480 BECAUSE INITIALLY YOU SHOWED AN 1402 00:49:58,480 --> 00:50:01,600 EAR INFECTION WHERE A LOT OF 1403 00:50:01,600 --> 00:50:03,640 NEUTROPHIL WAS MIGRATING BUT ARE 1404 00:50:03,640 --> 00:50:05,840 THERE ECM CHANGES WHICH ARE 1405 00:50:05,840 --> 00:50:06,200 SIMILAR OR? 1406 00:50:06,200 --> 00:50:08,200 >> SO WE HAVE LOOKED AT INFECTED 1407 00:50:08,200 --> 00:50:10,360 WOUNDS AND YOU STILL HAVE 1408 00:50:10,360 --> 00:50:11,600 ORGANIZED COLLAGEN IN AN 1409 00:50:11,600 --> 00:50:12,720 INFECTED WOUND AND IT'S A GOOD 1410 00:50:12,720 --> 00:50:14,360 QUESTION BECAUSE YOU HAVE TONS 1411 00:50:14,360 --> 00:50:15,760 OF MMPs AND THINGS THAT ARE 1412 00:50:15,760 --> 00:50:17,880 PRODUCED SO YOU MIGHT THINK THAT 1413 00:50:17,880 --> 00:50:21,000 YOU ACTUALLY WOULD LOSE THAT 1414 00:50:21,000 --> 00:50:21,600 ORGANIZED COLLAGEN, IT CHANGES 1415 00:50:21,600 --> 00:50:24,160 BUT YOU STILL HAVE ORGANIZED 1416 00:50:24,160 --> 00:50:24,520 COLA SHEN. 1417 00:50:24,520 --> 00:50:26,600 >> BUT NEUTROPHILS DON'T REALLY 1418 00:50:26,600 --> 00:50:27,080 CARE ABOUT IT. 1419 00:50:27,080 --> 00:50:28,800 >> THEY DON'T SEEM TO REALLY 1420 00:50:28,800 --> 00:50:30,200 CARE AND MACROPHAGES SEEM TO 1421 00:50:30,200 --> 00:50:31,200 CARE MORE. 1422 00:50:31,200 --> 00:50:33,760 >> THANK YOU SO MUCH. 1423 00:50:33,760 --> 00:50:34,280 >> YES. 1424 00:50:34,280 --> 00:50:36,840 >> I HAVE A QUESTION, SO--YOU 1425 00:50:36,840 --> 00:50:39,240 KNOW, CLINICALLY, WE HEAR A LOT 1426 00:50:39,240 --> 00:50:40,200 THESE DAYS, PROBABLY A LOT MORE 1427 00:50:40,200 --> 00:50:44,760 THAN IN THE PAST ABOUT 1428 00:50:44,760 --> 00:50:46,120 MACROPHAGE ACTIVATION SYNDROME 1429 00:50:46,120 --> 00:50:50,720 AND IN A WHOLE VARIETY OF 1430 00:50:50,720 --> 00:50:52,000 DISORDERS THAT ARE--I MEAN 1431 00:50:52,000 --> 00:50:53,640 THEY'RE ALL IMMUNE SYSTEM 1432 00:50:53,640 --> 00:50:56,320 DISORDERS BUT THEY'RE OTHERWISE 1433 00:50:56,320 --> 00:50:57,360 NOT PARTICULARLY MECHANISTICALLY 1434 00:50:57,360 --> 00:51:01,680 RELATED TO 1 ANOTHER, DO YOU 1435 00:51:01,680 --> 00:51:03,520 THINK--I MEAN, I GUESS I'M 1436 00:51:03,520 --> 00:51:06,080 HAVING TROUBLE DECIDING IF THE 1437 00:51:06,080 --> 00:51:07,400 NEUTROPHIL ACTIVATES THE 1438 00:51:07,400 --> 00:51:09,200 MACROPHAGE AND THE MACROPHAGE 1439 00:51:09,200 --> 00:51:10,720 EFFECTS ALL OF THE SYMPTOMS OR 1440 00:51:10,720 --> 00:51:14,280 IS IT THE OTHER WAY AROUND IS 1441 00:51:14,280 --> 00:51:16,920 THAT THE MACROPHAGE GETS 1442 00:51:16,920 --> 00:51:21,800 ACTIVATED AND THE BAD ACTOR IN 1443 00:51:21,800 --> 00:51:24,480 THESE DISORDER SYSTEM ACTUALLY 1444 00:51:24,480 --> 00:51:24,840 THE NEUTROPHIL? 1445 00:51:24,840 --> 00:51:26,040 >> YEAH, THAT'S A VERY GOOD 1446 00:51:26,040 --> 00:51:26,360 QUESTION. 1447 00:51:26,360 --> 00:51:29,280 I THINK OFTEN THE BAD ACTOR IS 1448 00:51:29,280 --> 00:51:32,280 THE NEUTROPHIL BUT OFTEN THE 1449 00:51:32,280 --> 00:51:33,320 MACROPHAGE IS ENCOURAGING THAT 1450 00:51:33,320 --> 00:51:36,160 BAD BEHAVIOR, SO THE PARTNERS IN 1451 00:51:36,160 --> 00:51:37,240 CRIME, AND ESSENCE, BUT I THINK 1452 00:51:37,240 --> 00:51:39,200 THERE'S NO QUESTION THAT 1453 00:51:39,200 --> 00:51:40,200 THERE'S--THIS IS CIRCULAR, 1454 00:51:40,200 --> 00:51:40,400 RIGHT? 1455 00:51:40,400 --> 00:51:42,600 AND THE THING THAT'S SO 1456 00:51:42,600 --> 00:51:44,640 COMPLICATED IS THAT NEUTROPHILS 1457 00:51:44,640 --> 00:51:46,520 ARE NOT ALL THE SAME, RIGHT? 1458 00:51:46,520 --> 00:51:49,000 THEY CAN BE GOOD OR BAD FOR 1459 00:51:49,000 --> 00:51:49,720 WOUND HEALING, CANCER, 1460 00:51:49,720 --> 00:51:52,200 MACROPHAGES ARE THE SAME WAY. 1461 00:51:52,200 --> 00:51:55,320 SO WHAT IT IS IS THE MORE WE'VE 1462 00:51:55,320 --> 00:51:58,920 LEARNED, THE MORE COMPLICATED, 1463 00:51:58,920 --> 00:52:00,560 YOU KNOW IT'S MORE HUMBLING, 1464 00:52:00,560 --> 00:52:02,400 DOES THAT MAKE SENSE? 1465 00:52:02,400 --> 00:52:11,280 THAT'S WHAT I'M FINDING BUT 1466 00:52:11,280 --> 00:52:11,480 YEAH. 1467 00:52:11,480 --> 00:52:13,160 >> YOU SHOWED THE STRUCTURALLY 1468 00:52:13,160 --> 00:52:15,720 ARE OF MIDGIFT, NO? 1469 00:52:15,720 --> 00:52:19,680 WHICH IS A C-TERMINAL AND IF IT 1470 00:52:19,680 --> 00:52:21,760 IS DEPLETED OR REMOVED IT DOES 1471 00:52:21,760 --> 00:52:24,240 NOT, IT HAS ITS EFFECT MY 1472 00:52:24,240 --> 00:52:26,600 QUESTION IS WHICH OF THESE 1473 00:52:26,600 --> 00:52:28,200 PARTNERS INTERACKING WITH THE 1474 00:52:28,200 --> 00:52:29,600 C-TERMINAL AND HAVE YOU HAD ANY 1475 00:52:29,600 --> 00:52:31,280 OF THEM? 1476 00:52:31,280 --> 00:52:34,920 FOR THE MYELOID DERIVED FACTOR. 1477 00:52:34,920 --> 00:52:39,120 >> SO YES, THEY BIND TO THE KDEL 1478 00:52:39,120 --> 00:52:40,760 ON THE C-TERMINUS OF THE MYELOID 1479 00:52:40,760 --> 00:52:43,600 GROWTH FACTOR SO, YOU KNOW WE 1480 00:52:43,600 --> 00:52:47,720 WILL HAVE TO SEE, WE THOUGHT OF 1481 00:52:47,720 --> 00:52:50,240 KDEL RECEPTOR AS BEING ER REDENT 1482 00:52:50,240 --> 00:52:52,480 RECEPTOR, AND THEN WHEN WE GOT 1483 00:52:52,480 --> 00:52:54,400 THE SEQ DATA AND THEY WERE IN 1484 00:52:54,400 --> 00:52:56,160 THE SAME EXACT NEUTROPHIL 1485 00:52:56,160 --> 00:53:01,840 SUBSETS AND UPREGULATED, I WENT 1486 00:53:01,840 --> 00:53:02,240 HUH. 1487 00:53:02,240 --> 00:53:04,040 MAYBE THIS IS ACTING AS A 1488 00:53:04,040 --> 00:53:05,120 RECEPTOR MORE BROADLY BECAUSE 1489 00:53:05,120 --> 00:53:06,400 THIS IS SHOWING THAT THE SECTORS 1490 00:53:06,400 --> 00:53:08,320 CAN TRAFFIC TO THE CELL SURFACE 1491 00:53:08,320 --> 00:53:10,920 SO THEN IT WOULD BE POSSIBLE 1492 00:53:10,920 --> 00:53:14,480 THAT THEY RECOGNIZE SECRETED 1493 00:53:14,480 --> 00:53:16,800 MYELOID DRIVE GROWTH FACTOR AND 1494 00:53:16,800 --> 00:53:18,040 THAT COULD DAMPEN NEUTROPHIL 1495 00:53:18,040 --> 00:53:20,200 RESPONSES DIRECTLY SO 1496 00:53:20,200 --> 00:53:21,640 IT'S--YEAH. 1497 00:53:21,640 --> 00:53:21,960 INTERESTING. 1498 00:53:21,960 --> 00:53:23,480 >> TELL BE NICE TO HAVE THIS 1499 00:53:23,480 --> 00:53:25,040 COMPLEX ISOLATED AND THEN STUDY 1500 00:53:25,040 --> 00:53:25,360 IT. 1501 00:53:25,360 --> 00:53:27,000 >> RIGHT. 1502 00:53:27,000 --> 00:53:27,200 RIGHT. 1503 00:53:27,200 --> 00:53:27,600 AND THAT'S--YEAH. 1504 00:53:27,600 --> 00:53:34,400 LOTS OF WORK TO DO. 1505 00:53:34,400 --> 00:53:35,600 >> VERY GOOD. 1506 00:53:35,600 --> 00:53:40,680 WELL, I THINK WE SHOULD THANK 1507 00:53:40,680 --> 00:53:41,600 DR. HUTTENLOCHER AGAIN FOR A 1508 00:53:41,600 --> 00:53:43,240 WONDERFUL SEMINAR AND A GREAT 1509 00:53:43,240 --> 00:53:44,920 DISCUSSION, THANK YOU ALL FOR 1510 00:53:44,920 --> 00:53:45,480 COMING. 1511 00:53:45,480 --> 00:53:47,560 >> THANK YOU FOR COME NOTHING 1512 00:53:47,560 --> 00:53:47,960 PERSON, GUYS. 1513 00:53:47,960 --> 00:00:00,000 >> VERY COOL TO SEE EVERYBODY.