1 00:00:04,998 --> 00:00:05,832 CAN EVERYBODY HEAR ME EMPLOY 2 00:00:05,832 --> 00:00:07,134 OKAY, WE WILL GET STARTED. 3 00:00:07,134 --> 00:00:08,702 SO IT'S A TRUE PLEASURE FOR ME 4 00:00:08,702 --> 00:00:10,404 TODAY TO INTRODUCE 1 OF OUR OWN 5 00:00:10,404 --> 00:00:13,306 AND ACTUALLY A COLLEAGUE OF MINE 6 00:00:13,306 --> 00:00:16,209 FROM THE NIDCR, THIERRY 7 00:00:16,209 --> 00:00:18,545 GAUTHIER, HE RECEIVED HIS DEGREE 8 00:00:18,545 --> 00:00:20,514 IN MOLECULAR BIOLOGY DURING 9 00:00:20,514 --> 00:00:21,548 TUMOR DEVELOP AM. 10 00:00:21,548 --> 00:00:25,085 HE LATER PURSUED A Ph.D. AT 11 00:00:25,085 --> 00:00:28,221 THE UNIVERSITY OF 12 00:00:28,221 --> 00:00:30,390 [INDISCERNIBLE] IN 13 00:00:30,390 --> 00:00:34,594 [INDISCERNIBLE] FRANCE STUDYING 14 00:00:34,594 --> 00:00:38,899 OUR PHAGOCYTOSIS COULD REGULATE 15 00:00:38,899 --> 00:00:40,400 INNATE IMMUNITY AND HE JOINED 16 00:00:40,400 --> 00:00:43,870 THE LAB AT NIH IN NIDECKER CR AT 17 00:00:43,870 --> 00:00:53,146 TWEBT 18 - 18 00:00:53,146 --> 00:00:55,082 -IN 2018, HE HAS FOCUSED ON 19 00:00:55,082 --> 00:00:57,284 THE ROLE OF MODULATING FUNCTIONS 20 00:00:57,284 --> 00:00:59,419 AND THE CONCEPT OF SEPSIS. 21 00:00:59,419 --> 00:01:02,389 HE ALSO STUDIED THE ROLE OF 22 00:01:02,389 --> 00:01:04,658 ANOTHER TGF SUPER FAMILY, ACT 95 23 00:01:04,658 --> 00:01:07,561 A AND HOW IT HAS MACROPHAGES 24 00:01:07,561 --> 00:01:08,728 DURING INFECTION TO IMMUNITY AND 25 00:01:08,728 --> 00:01:10,597 HE'S TRYING TO UNDERSTAND THE 26 00:01:10,597 --> 00:01:12,599 LINK BETWEEN TGF BETA AND, 27 00:01:12,599 --> 00:01:13,567 METABOLISM AND VARIOUS CONTEXT 28 00:01:13,567 --> 00:01:17,204 AND HE AIMS TO TARGET THESE 29 00:01:17,204 --> 00:01:20,107 PATHWAYS TO DEVELOP NEW 30 00:01:20,107 --> 00:01:21,108 THERAPEUTICS FOR AUTOINFECTION, 31 00:01:21,108 --> 00:01:21,675 IMMUNITY AND CANCER. 32 00:01:21,675 --> 00:01:27,414 IT'S I PLEASURE TO HAVE YOU 33 00:01:27,414 --> 00:01:27,614 TODAY. 34 00:01:27,614 --> 00:01:28,949 NTHANK YOU SO MUCH FOR THE 35 00:01:28,949 --> 00:01:29,316 INTRODUCTION. 36 00:01:29,316 --> 00:01:30,851 I WOULD LIKE ALSO TO THANK THE 37 00:01:30,851 --> 00:01:32,018 ORGANIZERS FOR GIVING ME THE 38 00:01:32,018 --> 00:01:37,290 OPPORTUNITY TO PRESENT THIS WORK 39 00:01:37,290 --> 00:01:39,126 EMPLOY SO I'M GOING TO JUMP 40 00:01:39,126 --> 00:01:42,829 RIGHT IN AND TODAY I WILL TALK 41 00:01:42,829 --> 00:01:44,831 ABOUT MACROPHAGES AND AS YOU 42 00:01:44,831 --> 00:01:48,235 KNOW, THEY ARE INNATE CELL 43 00:01:48,235 --> 00:01:48,535 TYPES. 44 00:01:48,535 --> 00:01:50,003 THEY ARE IMPORTANT TO HUMAN 45 00:01:50,003 --> 00:01:52,105 IMMUNE RESPONSE, SO AS AN 46 00:01:52,105 --> 00:01:53,707 EXAMPLE, THE MACROPHAGES IN THE 47 00:01:53,707 --> 00:01:55,041 LUNG WILL BE EXTREMELY IMPORTANT 48 00:01:55,041 --> 00:01:56,776 TO CLEAR THE PATHOGENS, 49 00:01:56,776 --> 00:01:57,611 MACROPHAGES IN THE LIMP FOADS 50 00:01:57,611 --> 00:01:59,913 WILL BE IMPORTANT TO HELP 51 00:01:59,913 --> 00:02:01,681 MOUNTING A PROPER ADAPTIVE 52 00:02:01,681 --> 00:02:03,283 IMMUNE RESPONSE BUT WHAT'S 53 00:02:03,283 --> 00:02:04,451 REALLY FASCINATING ABOUT THOSE 54 00:02:04,451 --> 00:02:06,686 SETS IS THAT THEY'RE ALSO VERY 55 00:02:06,686 --> 00:02:10,624 IMPORTANT TO MAINTAIN 56 00:02:10,624 --> 00:02:12,259 HOMEOSTASIS AND SO A BREAK IN 57 00:02:12,259 --> 00:02:14,094 THE FUNCTION COULD BE 58 00:02:14,094 --> 00:02:17,631 DELETERIOUS, AND SO IT'S VERY 59 00:02:17,631 --> 00:02:19,366 IMPORTANT TO CLEAR THE SEN 60 00:02:19,366 --> 00:02:20,000 ILLEGALSEN ENSEL THROUGH RED 61 00:02:20,000 --> 00:02:22,736 BLOOD CELLS EMPLOY SO IN OUR LAB 62 00:02:22,736 --> 00:02:28,708 WE ARE STUDYING THE ROLE OF 63 00:02:28,708 --> 00:02:29,309 TGFBETTA. 64 00:02:29,309 --> 00:02:32,379 BUT NOT THAT MUCH INNATE HUMAN 65 00:02:32,379 --> 00:02:33,880 RESPONSE SO I WILL JUST 66 00:02:33,880 --> 00:02:36,750 INTRODUCE A LITTLE BIT OF TGF 67 00:02:36,750 --> 00:02:37,284 BETA. 68 00:02:37,284 --> 00:02:39,085 BELONGS TO THE TGF BETA SUPER 69 00:02:39,085 --> 00:02:42,122 FAMILY OF PROTEIN AND IT'S 70 00:02:42,122 --> 00:02:44,457 COMPRISES SEVERAL MEMBERS LIKE 71 00:02:44,457 --> 00:02:47,460 THE BMPs, HERE, THE ACTINS AND 72 00:02:47,460 --> 00:02:50,463 THE TGF BETA IT'S THE MOST 73 00:02:50,463 --> 00:02:53,133 DRIEBED MEMBER OF THE TGF-B, AND 74 00:02:53,133 --> 00:02:55,202 IT'S IT NEEDS TO BE ACTIVATED 75 00:02:55,202 --> 00:02:59,573 AND THEN CAN BIND TO THE 76 00:02:59,573 --> 00:03:02,108 RECEPTORS TGF-B AND 2 IT WILL 77 00:03:02,108 --> 00:03:02,676 ACTIVATE DIFFERENT SIGNALING 78 00:03:02,676 --> 00:03:06,379 PATHWAYS SO THE MAIN SIGNATURES 79 00:03:06,379 --> 00:03:08,682 THAT WILLING PATHWAY THAT 80 00:03:08,682 --> 00:03:11,952 INVOLVES THE SMADS, THE 81 00:03:11,952 --> 00:03:14,187 ACTIVATION OF SMAT 2 AND 3, BIND 82 00:03:14,187 --> 00:03:16,823 TO SMAT 4, TRANSLOCATE TO THE 83 00:03:16,823 --> 00:03:20,360 NUCLEUS WRRKS IT CAN ACTIVATE 84 00:03:20,360 --> 00:03:22,963 REFRESESSED GENE SUPPRESS AND 85 00:03:22,963 --> 00:03:29,402 CAN BE SHARED BY ACTINS, TGF-B 86 00:03:29,402 --> 00:03:38,778 CAN ACTIVATE SIGNALING PATHWAYS. 87 00:03:38,778 --> 00:03:40,313 AND SO, THIS TGF BETA THAT I 88 00:03:40,313 --> 00:03:41,748 MENTIONED WE HAVE BEEN STUDYING 89 00:03:41,748 --> 00:03:45,385 IN THE LAB HAS BEEN DESCRIBED TO 90 00:03:45,385 --> 00:03:46,620 MODULATE IMMUNITY, OUR LAB BUT 91 00:03:46,620 --> 00:03:49,522 NOT ONLY HAS DESCRIBED THAT 92 00:03:49,522 --> 00:03:51,825 TGF BETA CAN INDUCE REGULATORY 93 00:03:51,825 --> 00:03:54,861 T-CELL GENERATION BUT ALSO CAN 94 00:03:54,861 --> 00:03:57,297 INDUCE TH17 AND TH9 CELLS AS 95 00:03:57,297 --> 00:04:01,201 WELL AS REPRESS TH1 AND 2 AND 96 00:04:01,201 --> 00:04:03,136 CDA CYTOTOXIC T-CELLS SO THIS IN 97 00:04:03,136 --> 00:04:04,838 THE CONTEXT OF IMMUNITY AND IN 98 00:04:04,838 --> 00:04:06,806 THE TERM OF INNATE IMMUNITY, 99 00:04:06,806 --> 00:04:09,676 MUCH LESS IS KNOWN, SPECIALLY IN 100 00:04:09,676 --> 00:04:11,678 THE CONTEXT OF MACROPHAGES, 101 00:04:11,678 --> 00:04:14,214 NEVERTHELESS, THEY ARE STILL 102 00:04:14,214 --> 00:04:18,385 PAPERS OUT THERE, THAT TGF BETA 103 00:04:18,385 --> 00:04:22,822 COULD BE A MODULATOR OF THE 104 00:04:22,822 --> 00:04:25,625 FUNCTION, THE TGF BETA PROMOTE 105 00:04:25,625 --> 00:04:26,559 ANTIINFLAMMATORY MACROPHAGES, IT 106 00:04:26,559 --> 00:04:27,193 CAN ALSO REGULATE MIRROR IMAGE 107 00:04:27,193 --> 00:04:29,863 IMRAIGZ EMPLOY IN THE CONTEXT OF 108 00:04:29,863 --> 00:04:31,197 HOMEOSTASIS, TGF BETA CAN 109 00:04:31,197 --> 00:04:32,365 PROPROTEIN COMPLEX TISSUE REPAIR 110 00:04:32,365 --> 00:04:33,633 AND THE RESOLUTION OF 111 00:04:33,633 --> 00:04:34,367 INFLAMMATION, SO I'VE BEEN 112 00:04:34,367 --> 00:04:36,369 STUDYING THAT QUITE A LOT DURING 113 00:04:36,369 --> 00:04:43,843 MY Ph.D., ESPECIALLY IN THE 114 00:04:43,843 --> 00:04:46,713 CONTEXT OF [INDISCERNIBLE], AND 115 00:04:46,713 --> 00:04:51,951 TGF BETA CAN ALSO CONTROL TISSUE 116 00:04:51,951 --> 00:04:53,253 REPAIR AND REGULATING MICROGLIA 117 00:04:53,253 --> 00:04:55,822 DEVELOPMENT AND FUNCTION AND 118 00:04:55,822 --> 00:04:59,259 FINALLY, TGF BETA SIGNALING IN 119 00:04:59,259 --> 00:05:01,594 MACROPHAGE SYSTEM DIFFERENT IN 120 00:05:01,594 --> 00:05:04,497 TUMOR DEVELOPMENT ESPECIALLY IN 121 00:05:04,497 --> 00:05:06,199 TUMOR METASTATIC DEVELOPMENT. 122 00:05:06,199 --> 00:05:07,667 SO THERE ARE A LOT OF QUESTIONS 123 00:05:07,667 --> 00:05:09,502 UNANSWERED TILL, SO IN A BETTER 124 00:05:09,502 --> 00:05:11,738 CONTEXT, TGF BETA IS NOT KNOWN 125 00:05:11,738 --> 00:05:14,808 DURING BACTERIA WILL INFECS. 126 00:05:14,808 --> 00:05:15,842 THE MOLECULAR MECHANISMS BY 127 00:05:15,842 --> 00:05:17,944 WHICH TGF BETA PLAYS IN INNATE 128 00:05:17,944 --> 00:05:18,812 CELLS IS NOT UNDERSTOOD. 129 00:05:18,812 --> 00:05:22,515 AND THE ROLE OF TGF BETA IN 130 00:05:22,515 --> 00:05:23,616 REGULATING MACROPHAGES IN 131 00:05:23,616 --> 00:05:25,218 METABOLISM IS BASKLY UNKNOWN. 132 00:05:25,218 --> 00:05:27,020 SO THAT WILL LEAD ME TO TALK 133 00:05:27,020 --> 00:05:29,255 ABOUT 2 PROGECS WE HAD IN THE 134 00:05:29,255 --> 00:05:33,126 LAB, THE FIRST 1 IS ABOUT HOW 135 00:05:33,126 --> 00:05:35,662 TGF BETA CAN CONTROL THIS IN 136 00:05:35,662 --> 00:05:37,831 METABOLISM IS THE SECOND 1 IS 137 00:05:37,831 --> 00:05:39,833 ABOUT THE ROLE AND ACTIVATION OF 138 00:05:39,833 --> 00:05:41,134 THE INFLAMMATION, SO I'M GOING 139 00:05:41,134 --> 00:05:44,237 TO START TALKING ABOUT IMMUNO 140 00:05:44,237 --> 00:05:45,872 METABOLISM AND SO, IMMUNO 141 00:05:45,872 --> 00:05:48,975 METABOLISM IS QUITE COMPLEX, 142 00:05:48,975 --> 00:05:50,643 IT'S INTRICATE PATHWAY THAT 143 00:05:50,643 --> 00:05:51,511 INTERACTS WITH 1 ANOTHER AND 144 00:05:51,511 --> 00:05:53,246 TODAY WE WILL TALK MORE 145 00:05:53,246 --> 00:05:54,080 SPECIFICALLY ABOUT THE 146 00:05:54,080 --> 00:05:56,950 GLYCOLYSIS, AND SO THE DPLI 147 00:05:56,950 --> 00:05:58,318 COLSIS START WITH THE BREATHER 148 00:05:58,318 --> 00:06:00,887 OF THE TBLUICOSE TO WHAT IS 149 00:06:00,887 --> 00:06:02,789 TRANSPORTERS, AND IT WILL BE 150 00:06:02,789 --> 00:06:04,224 DEGRADED BY DIFFERENT ENZYMES 151 00:06:04,224 --> 00:06:06,760 THAT WILL LEAD TO A GENERATION 152 00:06:06,760 --> 00:06:08,728 OF PYRUVATE AND IT HAS 2 CHOICES 153 00:06:08,728 --> 00:06:10,563 IT CAN ENTER THE MITOCHONDRIA 154 00:06:10,563 --> 00:06:12,699 AND FEED THE TISSUE CYCLE AS 155 00:06:12,699 --> 00:06:13,900 WELL AS THE [INDISCERNIBLE] AND 156 00:06:13,900 --> 00:06:19,672 IT CAN ALSO BE DEGRADED INTO THE 157 00:06:19,672 --> 00:06:22,509 GENERATION OF ENZYMES AND THIS 158 00:06:22,509 --> 00:06:23,576 IS CALLED AEROBIC FLI COLSIS 159 00:06:23,576 --> 00:06:24,944 COLSIS AND IT HAS BEEN 160 00:06:24,944 --> 00:06:25,645 DEMONSTRATED TO FOR THE 161 00:06:25,645 --> 00:06:26,646 IMPORTANT PATH WAIL IN THE SET 162 00:06:26,646 --> 00:06:28,615 WHEN IS THEY'RE ACTIVATED TO 163 00:06:28,615 --> 00:06:30,950 GENERATE ENERGY ESPECIALLY TO 164 00:06:30,950 --> 00:06:33,553 PROVIDE FAST GENERATION BUT ALSO 165 00:06:33,553 --> 00:06:35,021 BECAUSE ALL THOSE METABOLITES 166 00:06:35,021 --> 00:06:36,523 THEY CAN HAVE OTHER SIGNALING 167 00:06:36,523 --> 00:06:40,927 PATHWAYS AND THEY CAN ALSO FEED 168 00:06:40,927 --> 00:06:42,962 OTHER METABOLIC PATHWAYS. 169 00:06:42,962 --> 00:06:46,833 AND SO, THIS IS NOW VERY WELL 170 00:06:46,833 --> 00:06:48,201 STUDIED THAT CHANGES IN 171 00:06:48,201 --> 00:06:49,169 METABOLISM CAN AFFECT THE 172 00:06:49,169 --> 00:06:51,371 CHANGES IN HUMAN CELLS, 173 00:06:51,371 --> 00:06:52,472 ESPECIALLY MACROPHAGES AND WHAT 174 00:06:52,472 --> 00:06:56,209 IS THE CURRENT DOGMA, THAT WHEN 175 00:06:56,209 --> 00:06:56,643 MACROPHAGES BECOME 176 00:06:56,643 --> 00:06:57,377 PROINFLAMMATORY, ESPECIAL 177 00:06:57,377 --> 00:06:59,612 LEECIAL IN RESPONSE TO LPS AND 178 00:06:59,612 --> 00:07:00,780 INTERFEREON GAMMA SO THEY WILL 179 00:07:00,780 --> 00:07:03,450 START TO ENHANCE THE GLYCOLYSIS, 180 00:07:03,450 --> 00:07:05,318 THEY WILL HAVE A BROKEN TISSUE 181 00:07:05,318 --> 00:07:07,387 CYCLE AND A DECREASE 182 00:07:07,387 --> 00:07:08,288 [INDISCERNIBLE] ACTIVITY, SO 183 00:07:08,288 --> 00:07:09,022 MITOCHONDRIA RESPIRAIGZ AND THAT 184 00:07:09,022 --> 00:07:13,159 WILL HELP THEM SUSTAIN AN 185 00:07:13,159 --> 00:07:19,132 INFLAMMATORY PHENOTYPE FOR 186 00:07:19,132 --> 00:07:19,966 CYTOKINE, CHEMOKINES, MIGRATE, 187 00:07:19,966 --> 00:07:30,510 [INDISCERNIBLE] AND SO ON AND SO 188 00:07:38,751 --> 00:07:39,085 FORTH. 189 00:07:39,085 --> 00:07:43,690 SO THAT WILL HELP THEM SUSTAIN 190 00:07:43,690 --> 00:07:44,491 THEIR ENZYME INFLAMMATORY 191 00:07:44,491 --> 00:07:47,694 FUNCTION EMPLOY SO BASICALLY THE 192 00:07:47,694 --> 00:07:49,429 GLYCOLYSIS SUSTAINS INFLAMMATION 193 00:07:49,429 --> 00:07:49,796 IN MACROPHAGES. 194 00:07:49,796 --> 00:07:54,000 AND SO WE WANTED TO ASK A VERY 195 00:07:54,000 --> 00:08:01,140 SIMPLE QUESTION, WHICH IS BEST 196 00:08:01,140 --> 00:08:01,841 TGF-B TO REGULATE IMMUNE 197 00:08:01,841 --> 00:08:03,543 FUNCTION BECAUSE IT'S A VERY 198 00:08:03,543 --> 00:08:04,077 IMPORTANT QUESTION. 199 00:08:04,077 --> 00:08:05,678 SO WE STARTED WITH THE 200 00:08:05,678 --> 00:08:08,982 MACROPHAGES AND WE GIVE THEM 201 00:08:08,982 --> 00:08:12,151 TGF BETA AND WE RUN SPRMS. 202 00:08:12,151 --> 00:08:14,787 SO INTERESTINGLY WHEN WE GIVE 203 00:08:14,787 --> 00:08:16,356 TGF BETA TO MACROPHAGES NOT MUCH 204 00:08:16,356 --> 00:08:18,725 HAPPENED SO WE THOUGHT MAYBE WE 205 00:08:18,725 --> 00:08:20,460 MEDE TO ACTIVATE THOSE A LITTLE 206 00:08:20,460 --> 00:08:21,761 BIT LIKE T-CELLS ARE STIMULATED 207 00:08:21,761 --> 00:08:23,630 WITH ACTIVATION AND WE CHOOSE 208 00:08:23,630 --> 00:08:25,365 LPS WHICH IS A LIGAND THAT HAS 209 00:08:25,365 --> 00:08:27,901 BEEN VERY WELL KNOWN TO ACTIVATE 210 00:08:27,901 --> 00:08:32,405 TBLI COLSIS, SO I SHOULD GIVE 211 00:08:32,405 --> 00:08:34,741 LPS ENHANCE GLYCOLYSIS, BUT THEN 212 00:08:34,741 --> 00:08:35,542 EXTREMELY SURPRISINGLY TO US 213 00:08:35,542 --> 00:08:41,147 WHEN YOU GIVE THEM A COMBINATION 214 00:08:41,147 --> 00:08:43,850 OF TGF BETA THEY CAN FURTHER 215 00:08:43,850 --> 00:08:49,789 ENHANCE THE DPLI COLSIS, SO WE 216 00:08:49,789 --> 00:08:52,325 COME FROM THE ENDS WE OBSERVE 217 00:08:52,325 --> 00:08:57,330 THE UPTAKE OF GLUCOSE BY USING 218 00:08:57,330 --> 00:08:58,431 FLUORESCENT ANALOGUE, INCREASED 219 00:08:58,431 --> 00:09:00,233 AFTER TGF BETA TREATMENT IN 220 00:09:00,233 --> 00:09:04,170 THOSE MACROPHAGES AND THERE IS A 221 00:09:04,170 --> 00:09:05,572 REFLECT ANT IN THE SUPERINATENT, 222 00:09:05,572 --> 00:09:07,941 AND IT WAS UPREGULATED AFTER 223 00:09:07,941 --> 00:09:09,709 TGF BETA TREATMENT AND OVER ALL 224 00:09:09,709 --> 00:09:12,178 WE SHOW THAT TGF BETA CAN 225 00:09:12,178 --> 00:09:17,016 PROMOTE FLI COLSIS COLSIS AND 226 00:09:17,016 --> 00:09:17,684 MEDIATE REACTION. 227 00:09:17,684 --> 00:09:21,921 SO WE WANT TO ASK IS IT IN 228 00:09:21,921 --> 00:09:23,756 GENERAL WHEN MACROPHAGES ARE 229 00:09:23,756 --> 00:09:24,290 STIMULATED? 230 00:09:24,290 --> 00:09:27,293 SO WE STIMULATE WITH DIFFERENT 231 00:09:27,293 --> 00:09:30,396 TLR LIGANDS AND PREINFLAMMATORY 232 00:09:30,396 --> 00:09:33,499 CYTOKINES AND IT SHOULD GIVE 233 00:09:33,499 --> 00:09:34,334 MACROPHAGES LIKE 234 00:09:34,334 --> 00:09:35,501 [INDISCERNIBLE], YOU CAN ENHANCE 235 00:09:35,501 --> 00:09:38,338 TBLI COLSIS IN RESPONSE TO 236 00:09:38,338 --> 00:09:40,039 TGF BETA IT'S ALSO TRUE IN 237 00:09:40,039 --> 00:09:43,209 RESPONSE TO 848 WHICH IS A CLR7 238 00:09:43,209 --> 00:09:45,745 LIGAND AND IT'S ALSO TRUE WITH 239 00:09:45,745 --> 00:09:52,385 TNF ALPHA WITH IS DEMON TRAITING 240 00:09:52,385 --> 00:09:55,622 THAT TGF BETA AND THAT'S A 241 00:09:55,622 --> 00:09:56,122 GENERAL PHENOMENON. 242 00:09:56,122 --> 00:09:57,523 SO FOR THE FLI COLSIS COLSIS IS 243 00:09:57,523 --> 00:09:59,292 FINE, WHAT IS DIFFERENT, WHAT IS 244 00:09:59,292 --> 00:10:00,360 THE TYPE OF FUNCTION EMPLOY SO 245 00:10:00,360 --> 00:10:03,162 TO TRY TO UNDERSTAND THIS 246 00:10:03,162 --> 00:10:04,897 QUESTION, WE PERFORM RNASEQ 247 00:10:04,897 --> 00:10:07,533 ANALYSIS AND WE DID THAT IN 248 00:10:07,533 --> 00:10:09,836 COLLABORATION WITH THE DOCTOR 249 00:10:09,836 --> 00:10:13,673 AND, SO, AS CAN YOU OBSERVE 250 00:10:13,673 --> 00:10:15,808 HERE, WHEN YOU GIVE TGF BETA TOO 251 00:10:15,808 --> 00:10:20,813 MUCH IN RESPONSE TO LPS, IT 252 00:10:20,813 --> 00:10:21,481 DRAMATICALLY REMODELED THOSE 253 00:10:21,481 --> 00:10:23,049 MACROPHAGES AND IF YOU DEEPER IN 254 00:10:23,049 --> 00:10:24,684 TERMS WHAT HAVE GENES CHANGE, 255 00:10:24,684 --> 00:10:26,486 CAN YOU OBSERVE THERE ARE 256 00:10:26,486 --> 00:10:27,654 SEVERAL JOONS BELONGING TO DOWN 257 00:10:27,654 --> 00:10:30,423 REGULATION THAT ARE DOWN 258 00:10:30,423 --> 00:10:32,692 REGULATED LIKE CHEMOKINES AND 259 00:10:32,692 --> 00:10:35,261 FACTORS FOR EXAMPLE, WHILE IT 260 00:10:35,261 --> 00:10:37,563 PROMOTES EXPRESSION OF 261 00:10:37,563 --> 00:10:39,298 ANTIINFLAMMATORY FACERS LIKE 262 00:10:39,298 --> 00:10:47,907 TGF BETA I ALGENERATED ACE 1. 263 00:10:47,907 --> 00:10:49,776 THEY PROMOTE EXPRESSION OF 264 00:10:49,776 --> 00:10:50,410 RGENERATED ACE 1 AND 265 00:10:50,410 --> 00:10:52,945 DISCIPLINARY CREASE THE 266 00:10:52,945 --> 00:10:54,947 EXPRESSION OF DNF ALPHA SO DEMON 267 00:10:54,947 --> 00:10:56,716 TRAITING THE FACT THAT THE 268 00:10:56,716 --> 00:10:58,651 RESPONSE TO THE BETA THOSE 269 00:10:58,651 --> 00:11:00,787 ACTIVITIES MACROPHAGES HAVE ALSO 270 00:11:00,787 --> 00:11:02,488 DECREASED INFLATION IN RESPONSE 271 00:11:02,488 --> 00:11:03,756 TO THESE TGF BETA. 272 00:11:03,756 --> 00:11:06,559 SO, THE NEXT QUESTION, WE WANTED 273 00:11:06,559 --> 00:11:09,462 TO ADDRESS IS BY WHICH 274 00:11:09,462 --> 00:11:11,564 MECHANISMS, BEST PROMOTE TBLI 275 00:11:11,564 --> 00:11:12,098 COLSIS AND UNDERSTANDING A 276 00:11:12,098 --> 00:11:14,467 LITTLE BIT IN THE SIGNALING 277 00:11:14,467 --> 00:11:16,202 PATHWAY, AND SO IF I COME BACK 278 00:11:16,202 --> 00:11:18,171 TO THE EARLY DATA I JUST SHOWED 279 00:11:18,171 --> 00:11:20,406 YOU, WE NOTICE THAD 1 ENZYME 280 00:11:20,406 --> 00:11:24,243 BELONGS TO GLYCOLYSIS WHICH IS 281 00:11:24,243 --> 00:11:25,578 NAMED PFKL, AND THAT WAS THE 282 00:11:25,578 --> 00:11:29,015 ONLY 1 THAT CAME UP AND PFKL IS 283 00:11:29,015 --> 00:11:31,918 EXTREMELY IMPORTANT BECAUSE IT'S 284 00:11:31,918 --> 00:11:34,087 CATALYZING THE RATE IMAGING 285 00:11:34,087 --> 00:11:35,254 ENZYME THAT'S REVERSIBLE THAT 286 00:11:35,254 --> 00:11:38,424 LEADS TO THE GENERATION OF 287 00:11:38,424 --> 00:11:39,726 [INDISCERNIBLE] IN PHOSPHATE AND 288 00:11:39,726 --> 00:11:42,128 SO ISSUES THE [INDISCERNIBLE] 289 00:11:42,128 --> 00:11:43,229 PHOSPHATE WILL BE DIRECTED 290 00:11:43,229 --> 00:11:45,865 TOWARDS THE LOWER STEPS OF DPLI 291 00:11:45,865 --> 00:11:47,100 COLSIS, RATHER THAN BIOSYNTHESIS 292 00:11:47,100 --> 00:11:48,534 AND THE FLI COLSIS COSALATION 293 00:11:48,534 --> 00:11:49,569 PATHWAY. 294 00:11:49,569 --> 00:11:52,839 SO WE COME FROM AT THE MRNA 295 00:11:52,839 --> 00:11:55,374 LEVEL, THAT PFK WAS UPREGULATED 296 00:11:55,374 --> 00:11:56,809 AND WE COME AT THAT AT THE 297 00:11:56,809 --> 00:11:59,479 PROTEIN LEVEL AS WELL, AND THE 298 00:11:59,479 --> 00:12:01,948 ACTIVITY WAS ALSO UPREGULATED IN 299 00:12:01,948 --> 00:12:03,249 RESPONSE, DEMONSTRATING 300 00:12:03,249 --> 00:12:04,183 BASICALLY THAT TGF BETA CAN 301 00:12:04,183 --> 00:12:05,785 INDUCE THE EXPRESSION EMPLOY NOW 302 00:12:05,785 --> 00:12:08,855 DOES IT REGULATE DPLI COLSIS, 303 00:12:08,855 --> 00:12:11,390 SO, TO UNDERSTAND THAT POINT, WE 304 00:12:11,390 --> 00:12:14,494 USE AN SiRNA DREKED AGAINST 305 00:12:14,494 --> 00:12:16,863 PFKL EXPRESSION AND AS CAN YOU 306 00:12:16,863 --> 00:12:19,132 APPRECIATE HERE, IN THE CONTROL, 307 00:12:19,132 --> 00:12:21,434 TGF BETA IS ABLE TO PROMOTE MRI 308 00:12:21,434 --> 00:12:23,369 COLSIS BUT IT'S NOT THE CASE 309 00:12:23,369 --> 00:12:26,405 ANYMORE IN THE CASE OF SIRRNA, 310 00:12:26,405 --> 00:12:29,108 AND PREDICTION WHICH IS INCREASE 311 00:12:29,108 --> 00:12:33,146 UP TO TGF BETA BETA TREATMENT 312 00:12:33,146 --> 00:12:37,717 AND THE IT IS COMPLETELY 313 00:12:37,717 --> 00:12:37,984 ABROGATED. 314 00:12:37,984 --> 00:12:42,355 SO IT CAN INCREASE IN A PFKL 315 00:12:42,355 --> 00:12:42,822 MANNER. 316 00:12:42,822 --> 00:12:45,091 SO HOW DOES IT GET EXPRESSION? 317 00:12:45,091 --> 00:12:47,960 SO THE FIRST THING WE TRY TO 318 00:12:47,960 --> 00:12:49,495 ADDRESS IS WHETHER THERE IS 319 00:12:49,495 --> 00:12:50,563 INVOLVED BECAUSE AS I TOLD YOU 320 00:12:50,563 --> 00:12:53,699 IN THE INTRODUCTION, IT'S THE 321 00:12:53,699 --> 00:12:56,202 SIGNALING OF TGF BETA SO 3 322 00:12:56,202 --> 00:12:57,136 RECEPTORS WHICH WOULD INDUCE 323 00:12:57,136 --> 00:12:58,171 PHOSPHORYLATION AND ACTIVATION 324 00:12:58,171 --> 00:13:00,473 THAT CAN USE GENE EXPRESSION. 325 00:13:00,473 --> 00:13:05,645 BUT IF YOU--IF WE USE SMAD 3 326 00:13:05,645 --> 00:13:08,848 KNOCK OUT, TO INDUCE EXPRESSION 327 00:13:08,848 --> 00:13:11,384 IS ABSOLUTELY NOT CHANGED. 328 00:13:11,384 --> 00:13:13,886 AND THEN, THE FLI COLSIS COLSIS 329 00:13:13,886 --> 00:13:17,690 WITH THE CELLS, TGF BETA IS ABLE 330 00:13:17,690 --> 00:13:21,327 TO PROMOTE FLI COLSIS COLSIS, BY 331 00:13:21,327 --> 00:13:31,871 TGFBETTA IS INDEPENDENT OF SMAD 332 00:13:35,575 --> 00:13:37,743 3 EMPLOY SO WE LOOK FOR A FEW OF 333 00:13:37,743 --> 00:13:39,045 THEM BUT WE FIRST REALIZE THAT 334 00:13:39,045 --> 00:13:41,814 mTOR IS A VERY SPECIAL PART OF 335 00:13:41,814 --> 00:13:43,883 METABOLISM AND MACROPHAGES AND 336 00:13:43,883 --> 00:13:45,685 THAT CAN REGULATE INFLAMMATION 337 00:13:45,685 --> 00:13:46,786 AS WELL. 338 00:13:46,786 --> 00:13:48,754 AND THAT mTOR HAS BEEN 339 00:13:48,754 --> 00:13:50,790 REPORTED IN PAPERS TO BE ABLE TO 340 00:13:50,790 --> 00:13:53,259 BE ACTIVATED BY THE TGF BETA 341 00:13:53,259 --> 00:13:54,493 SIGNALING AND THE MACROPHAGES. 342 00:13:54,493 --> 00:13:57,330 SO THE WAY WE CAN MEASURE 343 00:13:57,330 --> 00:13:59,732 INTERACTION IS BY MEASURING 344 00:13:59,732 --> 00:14:02,902 PHOSPHORYLATION OF THE DOWN 345 00:14:02,902 --> 00:14:06,339 STREAM TARGETS OF S6/4 EBP1, SO 346 00:14:06,339 --> 00:14:10,243 AS YOU CAN SEE HERE, WE CAN 347 00:14:10,243 --> 00:14:12,044 PROMOTE PHOSPHORYLATION OF 6 AS 348 00:14:12,044 --> 00:14:12,845 WELL AS FOR EBP1. 349 00:14:12,845 --> 00:14:15,114 SO IF YOU LOOK AT THE ACTIVITY 350 00:14:15,114 --> 00:14:19,452 BY USING RAP O MICEIN WHICH IS 351 00:14:19,452 --> 00:14:29,962 AN INHIBITOR, CAN YOU ALSO GET 352 00:14:31,564 --> 00:14:35,534 THE PFKL, THE EXPRESSION IS 353 00:14:35,534 --> 00:14:36,102 DEPENDENT ON mTOR. 354 00:14:36,102 --> 00:14:38,804 SO NOW IT'S A KINASE, IT CANNOT 355 00:14:38,804 --> 00:14:39,872 DIRECTLY REGULATE GENE 356 00:14:39,872 --> 00:14:41,173 EXPRESSION, IT NEEDS A 357 00:14:41,173 --> 00:14:43,943 TRANSCRIPTION FACTOR BELOW, AND 358 00:14:43,943 --> 00:14:46,479 SO WE AGAIN TRIED DIFFERENT 359 00:14:46,479 --> 00:14:49,582 COMBINATIONS THAT CAME TO BE 360 00:14:49,582 --> 00:14:52,118 NEGATIVE WUWE TARTED TO GET 361 00:14:52,118 --> 00:14:53,719 INTERESTED IN [INDISCERNIBLE] SO 362 00:14:53,719 --> 00:14:57,623 AGAIN, IS SUPPOSED TO BE AN 363 00:14:57,623 --> 00:15:00,393 EXTREMELY REGULATOR OF THE 364 00:15:00,393 --> 00:15:01,360 METABOLISM ESPECIALLY 365 00:15:01,360 --> 00:15:02,295 MACROPHAGES AND SO WHAT WE COULD 366 00:15:02,295 --> 00:15:05,197 NOTICE AND THAT WAS A LITTLE BIT 367 00:15:05,197 --> 00:15:07,700 INTERESTING, THAT'S THE PROTEIN 368 00:15:07,700 --> 00:15:09,669 LEVELS OF AFFECTED BY TGF BETA 369 00:15:09,669 --> 00:15:12,605 BUT IT CAN BE PHOSPHORYLATED BY 370 00:15:12,605 --> 00:15:14,774 TGF BETA IT'S SERIES POINTSINE 371 00:15:14,774 --> 00:15:17,743 62 WHICH IS FORFORALATION AND 372 00:15:17,743 --> 00:15:21,580 THIS IS THE mTOR AND ABROGATED 373 00:15:21,580 --> 00:15:25,217 IN THE PRESENCE OF 374 00:15:25,217 --> 00:15:26,552 [INDISCERNIBLE] IT CAN REPRESENT 375 00:15:26,552 --> 00:15:29,689 THE TGF BETA IN THE SENSE AND IT 376 00:15:29,689 --> 00:15:33,559 WE DO COMPUTATIONAL ANALYSIS, 377 00:15:33,559 --> 00:15:35,161 THAT'S BINDING POTENTIAL BINDING 378 00:15:35,161 --> 00:15:40,666 SITE INTOS THE PFK PROMOTER AND 379 00:15:40,666 --> 00:15:41,901 IN RESPONSE TO TGF BETA SIERK 380 00:15:41,901 --> 00:15:43,202 AMERICA I CAN BIND TO THE 381 00:15:43,202 --> 00:15:46,505 PROMOTER AND THIS IS COMPLETELY 382 00:15:46,505 --> 00:15:50,109 OBLIGATIONS BROIGATED BY RAPP A 383 00:15:50,109 --> 00:15:53,279 MYOCIN, NOW IT'S RERESPONSIBLE 384 00:15:53,279 --> 00:15:55,648 FOR THE CHANGES IN EXPRESSION 385 00:15:55,648 --> 00:15:58,150 YES IT IS SO IF YOU USE A 386 00:15:58,150 --> 00:16:00,353 INHIBITOR, YOU CAN INVERT THE 387 00:16:00,353 --> 00:16:01,721 INDUCTION BY TGF BETA AND YOU 388 00:16:01,721 --> 00:16:03,923 CAN ALSO BLOCK THE INDUCTION OF 389 00:16:03,923 --> 00:16:06,258 GLYCOLYSIS IN THE PRESENCE OF 390 00:16:06,258 --> 00:16:08,327 THIS INHIBITOR, A DEMONITRATION 391 00:16:08,327 --> 00:16:11,497 THAT TGF BETA CAN REGULATE 392 00:16:11,497 --> 00:16:13,432 EXPRESSION, AND mTOR DEPENDENT 393 00:16:13,432 --> 00:16:13,699 MANNER. 394 00:16:13,699 --> 00:16:16,035 SO I SHOWED YOU THE SIGNALING BY 395 00:16:16,035 --> 00:16:20,373 WHICH IT CAN INDUCE PFK AND 396 00:16:20,373 --> 00:16:21,674 GLYCOLYSIS BUT I SHOWED YOU IN 397 00:16:21,674 --> 00:16:23,709 THE MEAN TIME, IT CAN DECREASE 398 00:16:23,709 --> 00:16:25,978 INFLAMMATION IN THE CELLS, SO IF 399 00:16:25,978 --> 00:16:27,146 THERE ARE ANY RELATIONSHIP 400 00:16:27,146 --> 00:16:28,948 BETWEEN THESE CHANGES IN 401 00:16:28,948 --> 00:16:31,217 GLYCOLYSIS AND INFLAMMATION, 402 00:16:31,217 --> 00:16:32,151 ACTUALLY, IT SEEMS THAT THERE 403 00:16:32,151 --> 00:16:35,454 ISN'T BECAUSE IF YOU USE THE 404 00:16:35,454 --> 00:16:38,090 PFKL THAT I CANNOT USE BEFORE, 405 00:16:38,090 --> 00:16:40,192 IF YOU REMEMBER, BUT THE ABILITY 406 00:16:40,192 --> 00:16:42,528 OF TGF BETA TO DECREASE ALPHA 407 00:16:42,528 --> 00:16:43,729 AND IL6 PRODUCTION, IS 408 00:16:43,729 --> 00:16:46,699 ABSOLUTELY NOT AFFECTED BY THE 409 00:16:46,699 --> 00:16:48,234 BLOCKADE OF PFK EXPRESSION, 410 00:16:48,234 --> 00:16:50,236 DEMONSTRATE THAGOREAN YOU HAVE A 411 00:16:50,236 --> 00:16:53,572 PROCESS, IN WHICH TGF BETA CAN 412 00:16:53,572 --> 00:16:54,840 PROMOTE GLYCOLYSIS AND DECREASE 413 00:16:54,840 --> 00:16:58,778 INFLAMMATION BUT THOSE PROCESSES 414 00:16:58,778 --> 00:16:59,712 ARE COMPLETELY UNCOUPLED TO 1 415 00:16:59,712 --> 00:17:01,714 ANOTHER EMPLOY SO HOW'S 416 00:17:01,714 --> 00:17:02,648 INFLAMMATION REGULATED THEN, WE 417 00:17:02,648 --> 00:17:04,283 CAME BACK TO SMAD 3 BECAUSE IT'S 418 00:17:04,283 --> 00:17:06,485 STILL VERY IMPORTANT FOR TGF 419 00:17:06,485 --> 00:17:15,995 BETA SIGNALING AND THIS TIME IF 420 00:17:15,995 --> 00:17:17,997 YOU USE THE MACROPHAGES AND 421 00:17:17,997 --> 00:17:20,666 INDUCE THOSE WHICH IS AN 422 00:17:20,666 --> 00:17:23,169 ANTIINFLAMMATORY MOLECULE IS 423 00:17:23,169 --> 00:17:24,470 COMPLETELY ABROGATED IN THE 424 00:17:24,470 --> 00:17:24,937 MACROPHAGES. 425 00:17:24,937 --> 00:17:28,607 SO HAVE YOU BASICALLY SHOWING 426 00:17:28,607 --> 00:17:30,709 THAT TGF BETA REGULATES SMAD 3 427 00:17:30,709 --> 00:17:34,246 AND YOU HAVE THIS UNCOUPLING, 428 00:17:34,246 --> 00:17:39,785 TGF BETA CO PROMOTE GLYCOLYSIS, 429 00:17:39,785 --> 00:17:42,788 BUT VERY NICE BUT THIS IS KNOW 430 00:17:42,788 --> 00:17:42,988 VITRO. 431 00:17:42,988 --> 00:17:44,690 WHAT'S HAPPENING IN VIVO. 432 00:17:44,690 --> 00:17:46,192 SO TROO TOY TO UNDERSTAND WHAT 433 00:17:46,192 --> 00:17:48,561 THE IS ROLE OF TGF BETA IN VIVO. 434 00:17:48,561 --> 00:17:51,897 WE WENT TO THE CONTEXT OF SEPSIS 435 00:17:51,897 --> 00:17:53,766 WHICH IS MIMICKED BY LPS 436 00:17:53,766 --> 00:17:56,869 INVITIO, AND WE USED AN LPS 437 00:17:56,869 --> 00:17:57,837 SEPSIS MODEL AND WE 438 00:17:57,837 --> 00:18:06,445 INRECYCLINGED IP INTO THE MICE 439 00:18:06,445 --> 00:18:09,782 AND THEN WE WORKED ON THE MODEL, 440 00:18:09,782 --> 00:18:13,319 THIS IS A SEPSIS MODEL FOR REAL 441 00:18:13,319 --> 00:18:14,987 INFECTION ONGOING TO THE PERO 442 00:18:14,987 --> 00:18:16,622 TONIGHTIS AND SO IF YOU INJECT 443 00:18:16,622 --> 00:18:18,057 LPS IN THE MICE, CAN YOU OBSERVE 444 00:18:18,057 --> 00:18:22,761 THAT IN THE BLOOD OF THOSE MICE, 445 00:18:22,761 --> 00:18:26,232 SO AT 6 HOURS NOT MUCH CHANGES 446 00:18:26,232 --> 00:18:31,003 BUT AT 16 HOURS, WE SEE SOME 447 00:18:31,003 --> 00:18:31,237 CHANGES. 448 00:18:31,237 --> 00:18:32,605 AND SO, BECAUSE THERE IS A LOT 449 00:18:32,605 --> 00:18:34,773 OF TGF BETA WE WANT TO BLOCK IT 450 00:18:34,773 --> 00:18:39,678 SO WHAT IS HAPPENING IF YOU 451 00:18:39,678 --> 00:18:42,581 BLOCK TGF BETA BY USING AN 452 00:18:42,581 --> 00:18:45,384 ANTIBODY ABOUT 15 MINUTES BEFORE 453 00:18:45,384 --> 00:18:45,784 INJECTION. 454 00:18:45,784 --> 00:18:48,821 IF YOU BLOCK TGF BETA YOU CAN 455 00:18:48,821 --> 00:18:50,890 PROTECT THE MICE OF THE MICE 456 00:18:50,890 --> 00:18:51,657 WHICH IS SURPRISING BECAUSE 457 00:18:51,657 --> 00:18:54,627 TGF BETA IS SUPPOSED TO BE 458 00:18:54,627 --> 00:18:56,896 ANTIINFLAMMATORY, SO WE EXPECTED 459 00:18:56,896 --> 00:19:00,232 THE OPPOSITE, SO JUST SHOWING 460 00:19:00,232 --> 00:19:02,668 THAT PROMOTE BLOCKING IN DURING 461 00:19:02,668 --> 00:19:05,304 SEPSIS COULD BE DELETERIOUS 462 00:19:05,304 --> 00:19:07,473 MAYBE, SO WE INJECT THOSE INTO 463 00:19:07,473 --> 00:19:09,108 THE MICE BEFORE LPS INJECTION 464 00:19:09,108 --> 00:19:14,013 AND THAT TIME WE INYECT INTO THE 465 00:19:14,013 --> 00:19:16,849 MICE YOU CAN ACTUALLY AND BUT IN 466 00:19:16,849 --> 00:19:18,250 TERM OF INFLAMMATION, THERE IS 467 00:19:18,250 --> 00:19:24,256 NO MAJOR CHANGES IF YOU LOOK AT 468 00:19:24,256 --> 00:19:27,059 THE LEVELS OF TNP ALPHA, SO 469 00:19:27,059 --> 00:19:28,460 DEMONSTRATING THAT IT IS DELETE 470 00:19:28,460 --> 00:19:29,929 ERIOUS FOR SEPSIS DEVELOPMENT 471 00:19:29,929 --> 00:19:35,834 BUT NOT BECAUSE IT CHANGES THE 472 00:19:35,834 --> 00:19:36,235 INPOLICEMANNATION. 473 00:19:36,235 --> 00:19:46,779 SO THEN IF IT'S NOT INFLAMMATION 474 00:19:48,781 --> 00:19:53,485 COULD BE AND IF YOU REMEMBER 475 00:19:53,485 --> 00:20:02,328 HERE, AND THEY ALSO HAVE 476 00:20:02,328 --> 00:20:04,730 INCREASED AND WHICH IS AN 477 00:20:04,730 --> 00:20:07,433 INHIBITOR TO KILL THE MICE WAS 478 00:20:07,433 --> 00:20:08,500 COMPLETELY ABROGATED AND MICE 479 00:20:08,500 --> 00:20:15,040 ARE PREDICTED FROM THERE. 480 00:20:15,040 --> 00:20:16,675 DEMONSTRATING THAT IT IS 481 00:20:16,675 --> 00:20:18,544 MEDIATED BY GLYCOLYSIS, SO HOW 482 00:20:18,544 --> 00:20:19,612 DOES GLYCOLYSIS REGULATE THE 483 00:20:19,612 --> 00:20:23,716 SURVIVAL OF THE MICE IN THIS 484 00:20:23,716 --> 00:20:26,385 CONTEXT IN SO WE TESTED A FEW 485 00:20:26,385 --> 00:20:27,419 PARAMETERS, LIKE TEMPERATURE, 486 00:20:27,419 --> 00:20:29,655 WEIGHT, MOST OF THEM CAME BACK 487 00:20:29,655 --> 00:20:32,958 NEGATIVE BUT WHAT WE NOTICED IS 488 00:20:32,958 --> 00:20:34,093 THAT THE COAGULATION AFTER THE 489 00:20:34,093 --> 00:20:37,296 TREATMENT AND THAT TGF BETA 490 00:20:37,296 --> 00:20:39,098 TREATMENT THEY HAD ENHANCED 491 00:20:39,098 --> 00:20:42,001 COAGULATION AND THIS IS VERY 492 00:20:42,001 --> 00:20:42,801 INTERESTING BECAUSE COAGULATION 493 00:20:42,801 --> 00:20:46,939 HAS BEEN DESCRIBED NOW VERY WELL 494 00:20:46,939 --> 00:20:48,774 DURING SEPSIS AND COVID-19 TO 495 00:20:48,774 --> 00:20:50,409 DRIVE THE DEC AND TO BE 496 00:20:50,409 --> 00:20:51,010 EXTREMELY IMPORTANT FOR THE 497 00:20:51,010 --> 00:20:55,781 DEATH OF THE PATIENTS. 498 00:20:55,781 --> 00:20:58,117 AND DISCOAGULATION WHICH IS 499 00:20:58,117 --> 00:20:59,652 ENHANCED BY TGF BETA SO 500 00:20:59,652 --> 00:21:00,819 ENHANCEMENT THAT THEY NEED LESS 501 00:21:00,819 --> 00:21:04,356 TIME TO CO AGULATE THE BLOOD, 502 00:21:04,356 --> 00:21:05,891 THAT'S WHY IT'S INCREASED 503 00:21:05,891 --> 00:21:07,826 BECAUSE IF IT'S TO THE 504 00:21:07,826 --> 00:21:08,827 [INDISCERNIBLE], THE INHIBITOR 505 00:21:08,827 --> 00:21:12,865 YOU CAN REVERT BACK TO THE 506 00:21:12,865 --> 00:21:14,833 TGF BETA EFFECT NORMAL LEVELS, 507 00:21:14,833 --> 00:21:20,239 AND WE ALSO USE HEP A RIN, TO 508 00:21:20,239 --> 00:21:21,640 BLOCK THE CO AGZULATION IN THOSE 509 00:21:21,640 --> 00:21:24,476 MICE IN TERMS OF SURVIVAL, WHY 510 00:21:24,476 --> 00:21:25,778 TGF BETA CAN INDUCE THE DEATH OF 511 00:21:25,778 --> 00:21:30,683 THE MICE, IT'S ABROGATED IF YOU 512 00:21:30,683 --> 00:21:34,887 TREAT THE MICE WITH HEPARIN. 513 00:21:34,887 --> 00:21:36,322 THIS IS AXIS THAT IS RESPONSIBLE 514 00:21:36,322 --> 00:21:39,325 FOR THE DEATH OF MICE IN THE 515 00:21:39,325 --> 00:21:39,658 SEPSIS MODEL. 516 00:21:39,658 --> 00:21:40,959 SO I WANTED TO TRY TO UNDERSTAND 517 00:21:40,959 --> 00:21:45,998 A LITTLE BIT MORE BY THESE 518 00:21:45,998 --> 00:21:46,765 MECHANISMS TO PROMOTE 519 00:21:46,765 --> 00:21:49,468 COAGULATION AND SO WE CAME BACK 520 00:21:49,468 --> 00:21:53,172 TO OUR RNA SEQ DATA AND 1 GENE 521 00:21:53,172 --> 00:21:55,040 BELONGS TO COAGULATION AND IT'S 522 00:21:55,040 --> 00:21:59,244 F13 A, AND SO, IT'S A VERY 523 00:21:59,244 --> 00:22:01,880 SIMPLELIFIED SCHEMATIC OF 524 00:22:01,880 --> 00:22:04,049 COAGULATION, BUT THE INTRINSIC 525 00:22:04,049 --> 00:22:04,883 AND EXTRINSIC PATHWAY WILL LEAD 526 00:22:04,883 --> 00:22:06,819 TO FIEB RIN AND IT WILL HAVE TO 527 00:22:06,819 --> 00:22:08,287 BE STABILIZED AND CROSS LINKED 528 00:22:08,287 --> 00:22:11,056 TO FORM A PROPER CLOTS TO 529 00:22:11,056 --> 00:22:12,324 LOCALIZE COAGULATION AND THIS IS 530 00:22:12,324 --> 00:22:15,527 THE ROLE OF F13 TO DISCOVER THIS 531 00:22:15,527 --> 00:22:15,994 CROSS LINK. 532 00:22:15,994 --> 00:22:20,165 AND IT COULD COME FROM IN 533 00:22:20,165 --> 00:22:21,967 MACROPHAGES IN VIVO IN THE 534 00:22:21,967 --> 00:22:23,502 SEPSIS MODEL, THOSE MACROPHAGES 535 00:22:23,502 --> 00:22:25,804 THEY HAVE INCREASED LEVEL OF F13 536 00:22:25,804 --> 00:22:29,141 EXPRESSION, AND THIS IS ALSO 537 00:22:29,141 --> 00:22:31,210 TRUE INVITIO AND THIS IS 538 00:22:31,210 --> 00:22:34,213 DEPENDENT ON TBLI COLSIS BECAUSE 539 00:22:34,213 --> 00:22:39,385 IF IT'S G, YOU COMPLETELY 540 00:22:39,385 --> 00:22:41,487 REVERSE THE EXPRESSION, THE 541 00:22:41,487 --> 00:22:43,956 INDUCTION OF 13 F A IS 542 00:22:43,956 --> 00:22:45,657 COMPLETELY ABROGATED. 543 00:22:45,657 --> 00:22:47,126 DEMONSTRATE THAGOREAN TGF BETA 544 00:22:47,126 --> 00:22:49,895 CAN PROMOTE EXPRESSION OF F13 A 545 00:22:49,895 --> 00:22:52,731 1 IN A DEPENDENT MANNER AND IT 546 00:22:52,731 --> 00:22:56,034 CAN DRIVEWAY THE COAGULATION 547 00:22:56,034 --> 00:22:56,402 INTO THOSE MICE. 548 00:22:56,402 --> 00:22:59,238 SO FINALLY, WE WANTED TO TRY TO 549 00:22:59,238 --> 00:23:00,439 UNDERSTAND WHETHER THIS PATHWAY 550 00:23:00,439 --> 00:23:02,307 COULD BE ACTIVE DURING HUMAN 551 00:23:02,307 --> 00:23:07,012 SEPSIS, AND SO, WE WERE ABLE TO 552 00:23:07,012 --> 00:23:09,648 INTERROGATE SIPPINGLE SET RNASEQ 553 00:23:09,648 --> 00:23:10,616 DATABASES FROM THE BROAD 554 00:23:10,616 --> 00:23:11,650 EN--STRATEGIESITUTE THAT HAD 555 00:23:11,650 --> 00:23:12,951 BEEN PUBLISHED AND SO HERE THIS 556 00:23:12,951 --> 00:23:16,688 IS A SEPSIS COHORT, SO THIS IS 557 00:23:16,688 --> 00:23:18,190 MACROPHAGES, MYELOID CELLS LET'S 558 00:23:18,190 --> 00:23:20,392 SAY TO BE MORE PRECISE AND SO 559 00:23:20,392 --> 00:23:22,795 THEY WERE ABLE TO GATHER 560 00:23:22,795 --> 00:23:26,165 DIFFERENT TYPE OF PEASHTS, 561 00:23:26,165 --> 00:23:29,334 HEALTHY VOLUNTEERS WITH LIFE 562 00:23:29,334 --> 00:23:31,170 INFECTION, NOT LIFE THREATENING 563 00:23:31,170 --> 00:23:34,072 SUCH AS UTI OR PATES THAT END UP 564 00:23:34,072 --> 00:23:36,041 IN THE E. R. AND HAVE SEPSIS, 565 00:23:36,041 --> 00:23:37,443 FOR THOSE PATES THAT HAVE SEPSIS 566 00:23:37,443 --> 00:23:42,247 THEY ARE EXPRESSION OF FTA 1 AND 567 00:23:42,247 --> 00:23:43,549 TGF BETA RECEPTOR 1, AND WE DID 568 00:23:43,549 --> 00:23:46,084 A SIMILAR ATHAT WILL SIS IN A 569 00:23:46,084 --> 00:23:48,187 COVID-19 COHORT BECAUSE COVID-19 570 00:23:48,187 --> 00:23:49,421 PATIENTS THEY BASIC LYE DIE OF 571 00:23:49,421 --> 00:23:51,723 SEPSIS AND WE COULD OBSERVE THAT 572 00:23:51,723 --> 00:23:56,261 IN MYELOID CELLS MAINLY, AFTER 573 00:23:56,261 --> 00:24:01,366 COVID-19, THEY HAVE EXPRESSION 574 00:24:01,366 --> 00:24:03,735 OF F13A, AND DEMON TRAITING THAT 575 00:24:03,735 --> 00:24:05,904 THIS DISCOAGULATION CAN BE USED 576 00:24:05,904 --> 00:24:09,508 BY TGF BETA AND BE ACTIVE DURING 577 00:24:09,508 --> 00:24:10,242 SEPSIS IN HUMAN. 578 00:24:10,242 --> 00:24:11,810 SO TO SUMMARIZE THIS FIRST PART 579 00:24:11,810 --> 00:24:15,981 OF MY TALK, WHAT I SHOWED YOU, I 580 00:24:15,981 --> 00:24:18,484 HOPE I CONVINCED YOU, TGF BETA 581 00:24:18,484 --> 00:24:22,654 IN MACROPHAGES IN CONTEXT OF 582 00:24:22,654 --> 00:24:23,956 INFLAMMATION, CAN INDUCE 583 00:24:23,956 --> 00:24:27,259 RECEPTORS AND THE ACTIVATION OF 584 00:24:27,259 --> 00:24:29,161 mTOR, AND SIMI, SIMI CAN BIND 585 00:24:29,161 --> 00:24:31,430 AND USE THIS EXPRESSION, WHICH 586 00:24:31,430 --> 00:24:32,698 CAN PROMOTE GLYCOLYSIS, IN THE 587 00:24:32,698 --> 00:24:35,400 MEAN TIME IN THE SAME CELLS BUT 588 00:24:35,400 --> 00:24:39,538 IN A COMPLETELY UNCOUPLED WAY, 589 00:24:39,538 --> 00:24:41,673 TASK CAN PROMOTE ACTIVATION, CAN 590 00:24:41,673 --> 00:24:42,374 DECREASE INFLAMMATION, AND SO 591 00:24:42,374 --> 00:24:45,644 YOU HAVE KINE OF A BALANCE WITH 592 00:24:45,644 --> 00:24:46,712 INCREASED COAGULATION AND 593 00:24:46,712 --> 00:24:48,714 INCREASED INFLAMMATION BUT THE 594 00:24:48,714 --> 00:24:56,021 INCREASED TBLI COLSIS, IN VIVO, 595 00:24:56,021 --> 00:24:58,524 OKAY, YEAH, THE INCREASED 596 00:24:58,524 --> 00:25:01,593 GLYCOLYSIS, WILL LEAD TO AN 597 00:25:01,593 --> 00:25:02,227 INCREASE COAGULATION THAT WOULD 598 00:25:02,227 --> 00:25:04,129 LEAD TO THE TEAGHT OF THE MICE 599 00:25:04,129 --> 00:25:05,197 DURING SEPSIS, SO WE JUST 600 00:25:05,197 --> 00:25:06,932 PUBLISHED THIS PAPER IN THE 601 00:25:06,932 --> 00:25:08,467 SIGNALING A KOWPY MONTHS AGO, SO 602 00:25:08,467 --> 00:25:10,402 IF YOU'RE INTERESTED TO READ THE 603 00:25:10,402 --> 00:25:11,870 FULL STORY, YOU ARE WELCOME TO 604 00:25:11,870 --> 00:25:12,404 DO SO. 605 00:25:12,404 --> 00:25:13,572 ASK THEN I'M GOING TO SWITCH TO 606 00:25:13,572 --> 00:25:15,073 THE SECOND PART OF MY TALK, 607 00:25:15,073 --> 00:25:17,476 WHICH IS ABOUT THE REGULATION OF 608 00:25:17,476 --> 00:25:20,145 SMAD 3 DURING INFLAMMATION. 609 00:25:20,145 --> 00:25:25,350 AND SO, JUST TO MENTION SO THIS 610 00:25:25,350 --> 00:25:26,552 IS UNPUBLISHED DAILY BASIS THEA 611 00:25:26,552 --> 00:25:28,787 SO PLEASE DO NOT SHARE. 612 00:25:28,787 --> 00:25:33,425 SO IF I COME BACK TO THE FIRST 613 00:25:33,425 --> 00:25:35,594 SLIDE I SHOWED YOU DURING THE 614 00:25:35,594 --> 00:25:36,628 INTRODUCTION, TGF BETA IS 615 00:25:36,628 --> 00:25:38,330 EXTREMELY WELL KNOWN TO INDUCE 616 00:25:38,330 --> 00:25:39,865 PHOSPHORYLATION, VERY WELL 617 00:25:39,865 --> 00:25:41,867 KNOWN, BUT WHAT IS HAPPENING IN 618 00:25:41,867 --> 00:25:44,369 THE CONTEXT IS ESPECIALLY 619 00:25:44,369 --> 00:25:46,271 INFLAMMATION, AND HOW SMAD 3 CAN 620 00:25:46,271 --> 00:25:47,472 BE REGULATED IN THIS CONTEXT 621 00:25:47,472 --> 00:25:51,610 REMAINS PRETTY MUCH LARGELY 622 00:25:51,610 --> 00:25:51,843 UNKNOWN. 623 00:25:51,843 --> 00:25:54,046 AND SO WE STUDIED BY DOING VERY 624 00:25:54,046 --> 00:25:57,683 BASIC EXPERIMENTS IN THE LAB, 625 00:25:57,683 --> 00:25:58,317 INVITRORY HARVESTED MACROPHAGES 626 00:25:58,317 --> 00:26:01,453 AND WE GAVE THEM LPS TO ACTIVATE 627 00:26:01,453 --> 00:26:03,822 THEM AND SO SURPRISINGLY TO US, 628 00:26:03,822 --> 00:26:06,658 IF YOU GIVE LPS TO MACROPHAGES, 629 00:26:06,658 --> 00:26:08,427 IT CAN ENHANCE THE 630 00:26:08,427 --> 00:26:11,496 PHOSPHORYLATION OF SMAD 3 IN THE 631 00:26:11,496 --> 00:26:12,831 C-TERMINAL LEVEL. 632 00:26:12,831 --> 00:26:16,368 THIS HAPPENS, THIS STARTS AT 4 633 00:26:16,368 --> 00:26:19,004 HOURS, PEAKS AT 6 AND COMES DOWN 634 00:26:19,004 --> 00:26:19,504 AFTER 24 HOURS. 635 00:26:19,504 --> 00:26:22,341 SO DEMRONITRATION THAT LPS CAN 636 00:26:22,341 --> 00:26:23,642 INDUCE PHOSPHORYLATION, IN THE 637 00:26:23,642 --> 00:26:26,311 C-TERMINAL DOMAIN AND SO, WHAT 638 00:26:26,311 --> 00:26:28,046 WE NOTICED IS THAT THIS ONLY 639 00:26:28,046 --> 00:26:29,581 HAPPENS AT 4 HOURS AND THAT 640 00:26:29,581 --> 00:26:30,849 BEFORE, SO SUGGESTING THAT IT'S 641 00:26:30,849 --> 00:26:32,651 NOT A DIRECT SIGNALING BUT IT'S 642 00:26:32,651 --> 00:26:33,685 A SECONDARY RESPONSE AND THAT 643 00:26:33,685 --> 00:26:37,522 YOU MIGHT NEED NEW PROTEIN, TO 644 00:26:37,522 --> 00:26:38,490 INDUCE THIS PHOSPHORYLATION, SO 645 00:26:38,490 --> 00:26:42,160 WE TRY TO INVESTIGATE THE LITTLE 646 00:26:42,160 --> 00:26:44,196 BIT MECHANISM BY WHICH THEY 647 00:26:44,196 --> 00:26:44,830 INDUCE PHOSPHORYLATION, SO FIRST 648 00:26:44,830 --> 00:26:47,099 WE PLOKED THE TRANSLATION OF THE 649 00:26:47,099 --> 00:26:50,969 PROTEIN AND WE USED THE SIGHTY 650 00:26:50,969 --> 00:26:54,506 HEXAMIDHERE AND IF YOU OBSERVE 651 00:26:54,506 --> 00:26:57,609 THE ABILITY FORTHE REGULATION IS 652 00:26:57,609 --> 00:26:58,777 COMPLETELY PROHIBITED WITH 653 00:26:58,777 --> 00:26:59,177 [INDISCERNIBLE]. 654 00:26:59,177 --> 00:27:05,484 YOU NEED NEW PROTEIN TO INDUCE 655 00:27:05,484 --> 00:27:07,819 SMAD 3 PHOSPHORYLATION, SO WE 656 00:27:07,819 --> 00:27:11,323 SCREEN FOR SOME TGF BETA MEMBERS 657 00:27:11,323 --> 00:27:14,526 BY QPC R, THE THEY ARE AFFECTED 658 00:27:14,526 --> 00:27:16,728 BY LPS TREATMENT AT EARLY TIME 659 00:27:16,728 --> 00:27:18,597 POINT, THE TGF BETA RESOAPT OR2 660 00:27:18,597 --> 00:27:21,800 IS A LITTLE BIT DECREASED AND 661 00:27:21,800 --> 00:27:24,503 THE LEVEL OF TGF BETA MRNA 662 00:27:24,503 --> 00:27:26,271 INCREASE AT 6 HOURS, THEN IT DID 663 00:27:26,271 --> 00:27:28,840 NOT FIT WITH THE KINETIC BECAUSE 664 00:27:28,840 --> 00:27:29,875 WE OBSERVE PHOSPHORYLATION AT 4 665 00:27:29,875 --> 00:27:32,678 HOURS ALREADY AND WE COULD 666 00:27:32,678 --> 00:27:34,079 NONAPOPTOTIC THE FIND TGF BETA 667 00:27:34,079 --> 00:27:35,881 IN THE SUPERINATE EPT OF THOSE 668 00:27:35,881 --> 00:27:36,181 MACROPHAGES. 669 00:27:36,181 --> 00:27:39,618 SO THEN WE USE THE DEFICIENT 670 00:27:39,618 --> 00:27:41,019 MACROPHAGES COMING FROM THE 671 00:27:41,019 --> 00:27:43,522 MICE, AND AS YOU CAN OBSERVE 672 00:27:43,522 --> 00:27:48,694 HERE IN THE WILD-TYPE 673 00:27:48,694 --> 00:27:50,228 MACROPHAGE, IT CAN INDUCE VERY 674 00:27:50,228 --> 00:27:52,464 WELL AND THEN WITH THE DEFESHT 675 00:27:52,464 --> 00:27:55,100 CELLS, LPS CAN DO IT EVEN MORE 676 00:27:55,100 --> 00:27:58,336 WHILE TGF BETA EFFECT IS 677 00:27:58,336 --> 00:27:59,137 DRAMATICALLY ABROGATED SO DEMON 678 00:27:59,137 --> 00:28:02,140 TRAITING THAT THE INDUCTION OF 679 00:28:02,140 --> 00:28:04,009 SMAD 3 PHOSPHORYLATION BOY LPS 680 00:28:04,009 --> 00:28:08,947 IS COMPLETELY INDEPENDENT OF TE 681 00:28:08,947 --> 00:28:09,514 SIGNALING PATHWAY. 682 00:28:09,514 --> 00:28:12,117 SO IT'S NOT TGF BETA, SO WHAT 683 00:28:12,117 --> 00:28:14,252 COULD INDUCE SMAD 3 684 00:28:14,252 --> 00:28:15,654 POSFORALATION IN RESPONSE TO LPS 685 00:28:15,654 --> 00:28:18,423 AND WHAT WE NOTICE INDEED A VERY 686 00:28:18,423 --> 00:28:22,094 EARLY TIME POINT AFTER LPS 687 00:28:22,094 --> 00:28:26,431 TREATMENT, THE MACROPHAGE ARE 688 00:28:26,431 --> 00:28:27,966 ACTIVATING A RECEPTOR, AND AS 689 00:28:27,966 --> 00:28:32,437 WELL AS [INDISCERNIBLE] AND WE 690 00:28:32,437 --> 00:28:34,005 ALSO INCREASED A PREDICTION IN 691 00:28:34,005 --> 00:28:35,874 THE SUPERINATENT OF THE CELLS, 692 00:28:35,874 --> 00:28:38,577 SPECIALLY TARTING AT 4 HOURS. 693 00:28:38,577 --> 00:28:40,679 SO DEMONSTRATING THAT LPS CAN 694 00:28:40,679 --> 00:28:42,180 PROMOTE THE INDUCTION OF THE 695 00:28:42,180 --> 00:28:43,715 PATHWAY, AND THIS IS INTERESTING 696 00:28:43,715 --> 00:28:44,516 BECAUSE IT'S BEEN DEMON TRAIT 697 00:28:44,516 --> 00:28:47,652 INDEED ALL CELL TYPES TO BE ABLE 698 00:28:47,652 --> 00:28:48,520 TO INDUCE SMAD 3 699 00:28:48,520 --> 00:28:49,688 PHOSPHORYLATION, SO THAT COULD 700 00:28:49,688 --> 00:28:50,856 BE THE TARGET WE'RE LOOKING FOR 701 00:28:50,856 --> 00:28:54,326 AND SO WE'RE FORTUNATE TO BE 702 00:28:54,326 --> 00:28:57,162 ABLE TO GET THE PLUCKS MICE FROM 703 00:28:57,162 --> 00:28:57,863 DR. FLORRIA AT COLUMBIA 704 00:28:57,863 --> 00:29:00,098 UNIVERSITY AND SO WE GENERATED 705 00:29:00,098 --> 00:29:02,434 THE CREE MICE THAT HAS 706 00:29:02,434 --> 00:29:04,970 DEFICIENCY OF ACR1 B IN 707 00:29:04,970 --> 00:29:06,138 MACROPHAGES AND THEN AS CAN YOU 708 00:29:06,138 --> 00:29:08,940 APPRECIATE HERE, IT'S VERY 709 00:29:08,940 --> 00:29:10,041 DRAMATIC, LPS CAN INDUCE THE 710 00:29:10,041 --> 00:29:12,644 PHOSPHORYLATION AND IT IS 711 00:29:12,644 --> 00:29:13,845 COMPLETELY ABROGATED WHEN THE 712 00:29:13,845 --> 00:29:16,348 ACTIVITY RECEPTOR IS DOWN. 713 00:29:16,348 --> 00:29:18,784 SO DEMON TRAITING THAT SMAD 3 714 00:29:18,784 --> 00:29:20,852 CAN INDUCE PHOSPHORYLATION IN A 715 00:29:20,852 --> 00:29:21,253 DEPENDENT MANNER. 716 00:29:21,253 --> 00:29:24,389 SO WE'RE TRYING TO UNDERSTAND A 717 00:29:24,389 --> 00:29:25,824 BIT MORE THE SIGNALING PATHWAYS 718 00:29:25,824 --> 00:29:27,926 BY WHICH IT HAPPENS, I WILL NOT 719 00:29:27,926 --> 00:29:29,027 GO INTO ALL THE DETAILS ABOUT 720 00:29:29,027 --> 00:29:32,831 YOU WE SHOW OF COURSE THAT IT 721 00:29:32,831 --> 00:29:35,667 CAN WORK WITH THE CLR RECEPTOR, 722 00:29:35,667 --> 00:29:38,470 SO IT'S NOT AFFECTED, IT'S NOT 723 00:29:38,470 --> 00:29:42,174 IMPORTANT, BUT IT IS DEPENDENT 724 00:29:42,174 --> 00:29:44,910 ON NYDATE, AND THE MYK KINASES, 725 00:29:44,910 --> 00:29:47,646 SO NOW AGAIN, THEY ARE KINASES 726 00:29:47,646 --> 00:29:49,047 SO THEY HAVE TRANSCRIPTION 727 00:29:49,047 --> 00:29:50,949 FACTORS SO WE TRY TO LOOK FOR 728 00:29:50,949 --> 00:29:54,686 THE TRAPGZ KRIPGZ FACTOR. 729 00:29:54,686 --> 00:29:55,854 WE CHECK THEMMER, THEY'RE NOT 730 00:29:55,854 --> 00:29:57,155 INVOLVED AND THEN WE TART 731 00:29:57,155 --> 00:29:59,090 STUDYING TO BE IN AT THAT TIME 732 00:29:59,090 --> 00:30:01,159 51st BECAUSE STAT 5 STUDIED TO 733 00:30:01,159 --> 00:30:03,094 BE PHOSPHORYLATED 2 HOURS AFTER 734 00:30:03,094 --> 00:30:05,497 LPS TREATMENT, SO THAT COULD FIT 735 00:30:05,497 --> 00:30:08,466 OUR KENETICS, AND ALSO BECAUSE 736 00:30:08,466 --> 00:30:10,435 AT THAT TIME 5 HAS SOME 737 00:30:10,435 --> 00:30:11,636 POTENTIAL BINDING SITES INTO THE 738 00:30:11,636 --> 00:30:14,272 ACTIVE PROMOTER AND IN RESPONSE 739 00:30:14,272 --> 00:30:16,675 TO LPS, STAT 5 CAN BIND TO THE 740 00:30:16,675 --> 00:30:17,976 ACTIVITY PROMOTER. 741 00:30:17,976 --> 00:30:20,312 THE DEMONITRATION THAT LPS CAN 742 00:30:20,312 --> 00:30:24,516 INHIBIT STAT 5 AND IF YOU USE 743 00:30:24,516 --> 00:30:25,917 STAT 5 MACROPHAGES, THE ABILITY 744 00:30:25,917 --> 00:30:29,287 OF LPS TO INDUCE ACTIVE 745 00:30:29,287 --> 00:30:30,055 EXPRESSION IS DRAMATICALLY 746 00:30:30,055 --> 00:30:32,858 REDUCED AND THE ABILITY TO 747 00:30:32,858 --> 00:30:36,862 INDUCE SMAD 3 PHOSPHORYLATION IS 748 00:30:36,862 --> 00:30:38,463 PRIMITIVE FOR REUSE, THIS CAN 749 00:30:38,463 --> 00:30:40,632 INCREASE EXPRESSION AND SMAD 750 00:30:40,632 --> 00:30:42,500 THROW PHOSPHORYLATION, FOR STAT 751 00:30:42,500 --> 00:30:43,201 5 DEPENDENT PATHWAY. 752 00:30:43,201 --> 00:30:45,904 SO, THIS IS THE MOLECULAR 753 00:30:45,904 --> 00:30:48,874 MECHANISM, WHAT IS THE FUNCTION 754 00:30:48,874 --> 00:30:53,578 OF THIS SMAD 3 PATHWAY. 755 00:30:53,578 --> 00:30:57,182 SO IF YOU KNOCK DOWN THE 756 00:30:57,182 --> 00:31:01,119 MACROPHAGES, YOU HAVE INCREASED 757 00:31:01,119 --> 00:31:02,754 PRODUCTION OF IL6, VERY DRAMATIC 758 00:31:02,754 --> 00:31:04,623 AND LARGELY DECREASED EXPRESSION 759 00:31:04,623 --> 00:31:07,993 OF TGP BETA EYE WHICH IS AN 760 00:31:07,993 --> 00:31:08,727 ANTIINFLAMMA ARE TOY FACTOR. 761 00:31:08,727 --> 00:31:11,263 IF YOU DO THE SAME THING WITH 762 00:31:11,263 --> 00:31:13,765 GUT MACROPHAGES YOU HAVE THE 763 00:31:13,765 --> 00:31:16,635 SAME THING, SO IS THE WHYED THAT 764 00:31:16,635 --> 00:31:18,270 YOU CAN HAVE NATURAL BRICK TO 765 00:31:18,270 --> 00:31:19,237 INFLATION THAT IS PUT IN PLACE 766 00:31:19,237 --> 00:31:21,573 BY THE CELLS TO ARK VOID THE 767 00:31:21,573 --> 00:31:25,243 OVER ACTIVATED, THEY WOULD START 768 00:31:25,243 --> 00:31:26,711 TO PRODUCE ACTIVITY A THAT 769 00:31:26,711 --> 00:31:28,513 PRODUCE THE PHOSPHORYLATION BUT 770 00:31:28,513 --> 00:31:29,814 WILL DOWN REGULATE THE 771 00:31:29,814 --> 00:31:32,217 PHOSPHORYLATION, SO WE WANTED TO 772 00:31:32,217 --> 00:31:33,718 UNDERSTAND A LITTLE BIT DEEPER 773 00:31:33,718 --> 00:31:36,254 WHAT COULD BE THE MECHANISM BY 774 00:31:36,254 --> 00:31:37,422 WHICH THESE CHANGE VS BEEN AND 775 00:31:37,422 --> 00:31:41,726 SO WE PERFORM SOME RNASEQ ANINAL 776 00:31:41,726 --> 00:31:43,228 SIS IN THE SMAD 3 MACROPHAGES 777 00:31:43,228 --> 00:31:45,096 WITH THE LPS SO IT'S NOT 778 00:31:45,096 --> 00:31:47,065 SURPRISING I SHOWED NUCLEOTIDES 779 00:31:47,065 --> 00:31:48,733 RESPONSE TO LPS, THEY HAVE 780 00:31:48,733 --> 00:31:53,171 INCREASED YEENS BELONGS TO 781 00:31:53,171 --> 00:31:53,905 INFLAMMATION, IL6, CD64, AND SO 782 00:31:53,905 --> 00:31:58,076 ON AND SO FORTH AND THEY HAVE 783 00:31:58,076 --> 00:32:02,681 JEERPS RELATED TO ANTIINFLMATORY 784 00:32:02,681 --> 00:32:02,914 FACTORS. 785 00:32:02,914 --> 00:32:05,183 SO WE CAN SEE AT THE PATHWAY 786 00:32:05,183 --> 00:32:06,484 LEVEL, BUT WHAT'S VERY 787 00:32:06,484 --> 00:32:07,886 INTERESTING THAT IF YOU LOOK AT 788 00:32:07,886 --> 00:32:09,988 THE DOWN REGUTED PATHWAY THIS 789 00:32:09,988 --> 00:32:12,390 TIME, NOT UPREGULATED YOU CAN 790 00:32:12,390 --> 00:32:13,959 OBSERVE THAT MANY OF THIS 791 00:32:13,959 --> 00:32:18,229 PATHWAY, MOST OF THEM BELONG TO 792 00:32:18,229 --> 00:32:18,530 METABOLISM. 793 00:32:18,530 --> 00:32:21,232 ESPECIALLY METABOLISM OF LIPIDS, 794 00:32:21,232 --> 00:32:21,900 CARBOHYDRATES, METABOLISM AND SO 795 00:32:21,900 --> 00:32:23,034 ON AND SO FORTH. 796 00:32:23,034 --> 00:32:24,269 SO SUGGESTING THAT THERE COULD 797 00:32:24,269 --> 00:32:27,205 BE LIKE A METABOLIC REGULATION 798 00:32:27,205 --> 00:32:31,142 IN THE CELLS, AND WAWE COULD 799 00:32:31,142 --> 00:32:34,012 OBSERVE NOTABLY, THE 800 00:32:34,012 --> 00:32:36,748 MITOCHONDRIA METABOLISM COULD BE 801 00:32:36,748 --> 00:32:38,116 THOSE METABOLISM, GENERATION, 802 00:32:38,116 --> 00:32:40,752 FUNCTION, THEY ARE ALL 803 00:32:40,752 --> 00:32:44,823 NONREGULATED, AND BY DOING FAST 804 00:32:44,823 --> 00:32:45,690 STAINING FROM MIGHT O TRACKER 805 00:32:45,690 --> 00:32:47,993 WHICH IS A WAY TO STAIN 806 00:32:47,993 --> 00:32:49,461 MITOCHONDRIA, WE CAN PRESERVE 807 00:32:49,461 --> 00:32:51,596 THAT AND WE HAVE DECREASED 808 00:32:51,596 --> 00:32:53,064 MITOCHONDRIA LEVELS, AND THIS IS 809 00:32:53,064 --> 00:32:55,600 EVEN MORE TRUE IN PRESENCE OF 810 00:32:55,600 --> 00:32:58,203 MACROPHAGES AND WE HAVE THE SAME 811 00:32:58,203 --> 00:33:02,140 RESULTS IN PRESENCE OF ACGR 1 812 00:33:02,140 --> 00:33:02,841 BEING THE MACROPHAGE, 813 00:33:02,841 --> 00:33:04,976 DEMONSTRATING THAT THE PATHWAY 814 00:33:04,976 --> 00:33:06,444 SUPPORT THE BIODPEN SIS, THAT 815 00:33:06,444 --> 00:33:08,279 COULD BE VERY IMPORTANT BECAUSE 816 00:33:08,279 --> 00:33:12,784 MITOCHONDRIA IS A HUB TO CONTROL 817 00:33:12,784 --> 00:33:13,318 METABOLISM, INFLAMMATION, 818 00:33:13,318 --> 00:33:14,552 MACROPHAGES FUNCTION AND SO, WE 819 00:33:14,552 --> 00:33:16,654 TRY TO GO A BIT FURTHER AND WHAT 820 00:33:16,654 --> 00:33:19,057 YOU COULD OBSERVE IS THAT THOSE 821 00:33:19,057 --> 00:33:21,226 CHANGES IN MITOCHONDRIA LEVELS 822 00:33:21,226 --> 00:33:22,827 WILLING TO DECREASE LEVELS OF 823 00:33:22,827 --> 00:33:25,130 ATP, WE HAVE LESS ATP INTO THE 824 00:33:25,130 --> 00:33:28,400 CELLS AND DEMON TRAITING THIS 825 00:33:28,400 --> 00:33:30,568 ACTIVITY A INDUCTION PATHWAY CAN 826 00:33:30,568 --> 00:33:33,538 LEAD TO MITOCHONDRIA BIOGENESIS 827 00:33:33,538 --> 00:33:35,640 AND GENERATION OF ATP, SO I 828 00:33:35,640 --> 00:33:37,542 REALLY WANT TO EMPHASIZE THAT 829 00:33:37,542 --> 00:33:39,477 THIS NOT LIKE EXTREME LEVELS OF 830 00:33:39,477 --> 00:33:41,479 ATP, IT'S TRY TO MAINTAIN HOMEIO 831 00:33:41,479 --> 00:33:42,981 STASE EXPIS HAVE NORMAL LEVELS 832 00:33:42,981 --> 00:33:44,916 OF ATISSUING P WHICH NORMAL IN 833 00:33:44,916 --> 00:33:48,953 THE CELLS TO FUNCTION. 834 00:33:48,953 --> 00:33:52,690 SO, YOU HAVE DECREASED ATP, SO 835 00:33:52,690 --> 00:33:57,695 DO THOSE LEVELS SUSTAIN 836 00:33:57,695 --> 00:33:58,530 INFLAMMATION, SO WE TRY TO SEN 837 00:33:58,530 --> 00:34:00,398 ILLEGALSEN BACK TO THE CELLS SO 838 00:34:00,398 --> 00:34:02,467 WE USE THE ATP TO INDUCE 839 00:34:02,467 --> 00:34:05,703 ACTIVATION OF COURSE, AND SO, 840 00:34:05,703 --> 00:34:07,205 WHAT YOU COULD OBSERVE IN THE 841 00:34:07,205 --> 00:34:11,209 MACROPHAGES IN PRESENCE OF LPS, 842 00:34:11,209 --> 00:34:13,745 AND LEVELS OF ATP, CAN YOU HAVE 843 00:34:13,745 --> 00:34:17,949 THE LEVEL OF EXPRESSION WHICH IS 844 00:34:17,949 --> 00:34:18,983 AN ANTIINFLAMMATORY FACTOR 845 00:34:18,983 --> 00:34:22,320 AGAIN, THIS IS TRUE IN 846 00:34:22,320 --> 00:34:23,621 [INDISCERNIBLE] MACROPHAGES. 847 00:34:23,621 --> 00:34:25,023 SO DEMONSTRATING AGAIN, HAVE YOU 848 00:34:25,023 --> 00:34:25,957 THE PATHWAY, THIS ACTIVITY IN 849 00:34:25,957 --> 00:34:30,361 THE PATHWAY THAT CAN LEAD TO 850 00:34:30,361 --> 00:34:31,629 MITOCHONDRIA BIOGENESIS, NO MORE 851 00:34:31,629 --> 00:34:42,173 PREDICTION THAT CAN PROMOTE THE 852 00:34:44,409 --> 00:34:51,382 EXPANSION OF ANTIINFLAMMATORY 853 00:34:51,382 --> 00:34:51,716 EFFECTS. 854 00:34:51,716 --> 00:34:56,688 ATP CAN CONVERGE INTO ADP AND 855 00:34:56,688 --> 00:35:01,559 AMP BY C13 AND 39, AND THEN ADEN 856 00:35:01,559 --> 00:35:03,995 O SEEN CAN INDUCE RECEPTORS AND 857 00:35:03,995 --> 00:35:06,631 E DUCE SIGNALING WITH KREB, 858 00:35:06,631 --> 00:35:07,999 THAT'S OUR ANTIFLAMMATORY. 859 00:35:07,999 --> 00:35:10,668 IS IT THE CASE IN OUR SETTINGS, 860 00:35:10,668 --> 00:35:13,171 SO WE TRY TO INTERROGATE THAT, 861 00:35:13,171 --> 00:35:17,008 AND SO THE FIRST THING WE 862 00:35:17,008 --> 00:35:19,110 OBSERVE IS THAT LPS CAN 863 00:35:19,110 --> 00:35:20,945 DRAMATICALLY ENHANCE THE 864 00:35:20,945 --> 00:35:22,947 ACTIVATION OF CD73 AND THIS IS 865 00:35:22,947 --> 00:35:24,382 DEPENDENT ON SMAD 3, SO THE 866 00:35:24,382 --> 00:35:26,317 LEVELS ARE LARGELY DECREASED, 867 00:35:26,317 --> 00:35:30,822 AND ON THE ACT VIN A EXPRESSION, 868 00:35:30,822 --> 00:35:33,725 SO IT COMES BACK DOWN IN 869 00:35:33,725 --> 00:35:34,926 [INDISCERNIBLE] KNOCK OUT 870 00:35:34,926 --> 00:35:36,895 MACROPHAGES, SO DEMONITRATION 871 00:35:36,895 --> 00:35:38,863 THAT THIS CAN PROMOTE THE 872 00:35:38,863 --> 00:35:40,665 EXPRESSION OF CD7. 873 00:35:40,665 --> 00:35:42,867 AND SO NOW WE HAVE 874 00:35:42,867 --> 00:35:43,434 FUNCTIONALITY. 875 00:35:43,434 --> 00:35:44,903 SO IF YOU RESORT BACK THE LEVELS 876 00:35:44,903 --> 00:35:51,576 OF ATP AS I SHOWED YOU, YOU CAN 877 00:35:51,576 --> 00:35:54,245 PROMOTE THIS AND IF YOU PLOK IT, 878 00:35:54,245 --> 00:35:55,713 DOWN STREAM TRANSCRIPTION FACTOR 879 00:35:55,713 --> 00:35:57,215 OF KREB, CAN YOU ABNORMALITIES 880 00:35:57,215 --> 00:35:58,850 BRO IMAIT THE ATP EFFECT, THIS 881 00:35:58,850 --> 00:36:00,818 IS TRUE IN THE GUT MACROPHAGES 882 00:36:00,818 --> 00:36:11,362 AND THIS IS ALSO TRUE IN THE--WE 883 00:36:14,999 --> 00:36:16,634 CAN BIND TO BELONGING TO 884 00:36:16,634 --> 00:36:19,604 MITOCHONDRIA FUNCTION WHICH CAN 885 00:36:19,604 --> 00:36:21,105 PROMOTE MITOCHONDRIA BIOGENESIS, 886 00:36:21,105 --> 00:36:24,008 THE SYNTHASE OF ATP AT NORMAL 887 00:36:24,008 --> 00:36:26,010 HOMEOSTATIC LEVELS WHICH CAN BE 888 00:36:26,010 --> 00:36:27,712 CONVERTED JUST IN CD73 THAT CAN 889 00:36:27,712 --> 00:36:30,181 CIRCLE BACK INTO THE CELLS, 890 00:36:30,181 --> 00:36:36,254 ACTIVATE KREB AND IN THE WITH 891 00:36:36,254 --> 00:36:40,792 SMAD 3 CAN WORK ON THE FOR 892 00:36:40,792 --> 00:36:41,025 EXAMPLE. 893 00:36:41,025 --> 00:36:43,127 SO THIS IS THE MECHANISMS 894 00:36:43,127 --> 00:36:44,395 INVITRO, NOW WHAT IS HAPPENING 895 00:36:44,395 --> 00:36:46,764 IN VIVO IS THE PATHWAY IS STILL 896 00:36:46,764 --> 00:36:49,133 ACTIVE, AS IT PLAYS, SO WE CAME 897 00:36:49,133 --> 00:36:54,839 BACK TO OUR LPS, WE WE INYEBED 898 00:36:54,839 --> 00:36:57,342 LPS INTO THE MICE, WE ISOLATED 899 00:36:57,342 --> 00:36:58,509 THE MACROPHAGES 6 HOURS AFTER 900 00:36:58,509 --> 00:36:59,477 THE INJECTION OF THE DISEASE AND 901 00:36:59,477 --> 00:37:00,912 WE LOOKEDDA THE THIS PATHWAY AND 902 00:37:00,912 --> 00:37:02,113 IT WAS ACTIVE SO AGAIN, I'M 903 00:37:02,113 --> 00:37:04,682 GOING TO SHOW YOU THE LPS DATA 904 00:37:04,682 --> 00:37:07,085 BUT WE HAVE SAME DATA WITH THE 905 00:37:07,085 --> 00:37:11,055 SEPSIS MODEL WHERE WE HAVE REAL 906 00:37:11,055 --> 00:37:12,991 INFECTION ONGOING. 907 00:37:12,991 --> 00:37:19,664 AND SO IN THOSE MICE IN INCREASE 908 00:37:19,664 --> 00:37:20,898 THE PHOSPHORYLATION IN RESPONSE 909 00:37:20,898 --> 00:37:24,435 TO LPS, IN THE SEPSIS MODEL, SO 910 00:37:24,435 --> 00:37:26,404 IT'S PHOSPHORYLATED, AND THEN, 911 00:37:26,404 --> 00:37:29,407 THIS IS HAPPENING MANICALLY ON 912 00:37:29,407 --> 00:37:30,541 MACROPHAGES, SO WOE HAVE 913 00:37:30,541 --> 00:37:32,477 INCREASED FOCUSED ONS FORALATION 914 00:37:32,477 --> 00:37:34,779 OF MACROPHAGES BUT NOT THAT MUCH 915 00:37:34,779 --> 00:37:36,748 INTO THE NUTRIFILL OR A LITTLE 916 00:37:36,748 --> 00:37:38,716 BIT WITH THE B-EABT CELLS AND 917 00:37:38,716 --> 00:37:41,486 THEN, IS IT DEPENDENT ON TGF 918 00:37:41,486 --> 00:37:43,521 BETA OR ACT VIN, WHAT WE COULD 919 00:37:43,521 --> 00:37:45,189 OBSERVE AS CAN YOU OBSERVE IN 920 00:37:45,189 --> 00:37:48,860 THIS GRAPH, SO IN WILD-TYPE, 921 00:37:48,860 --> 00:37:50,461 MICE, THE MACROPAGES DO NOT 922 00:37:50,461 --> 00:37:51,996 REALLY HAVE ANY SMAD 3 923 00:37:51,996 --> 00:37:52,830 PHOSPHORYLATION WHICH MAKES 924 00:37:52,830 --> 00:37:55,066 SENSE IN RESPONSE OF LPS, THIS 925 00:37:55,066 --> 00:37:59,270 IS PLURIBU CURVE, INCREASED BUT 926 00:37:59,270 --> 00:38:02,307 IN THE TGF BETA MICE, THEY CAN 927 00:38:02,307 --> 00:38:03,074 STILL INCREASE THE 928 00:38:03,074 --> 00:38:04,475 PHOSPHORYLATION AND THERE IS 929 00:38:04,475 --> 00:38:06,744 ABSOLUTELY NO DIFFERENCE. 930 00:38:06,744 --> 00:38:10,982 SO DEMON TRAITING THAT LPS CAN 931 00:38:10,982 --> 00:38:13,751 WORK IN VIVO DURING SEPSIS AND 932 00:38:13,751 --> 00:38:18,690 SO IS IT DEPENDENT ON THE 933 00:38:18,690 --> 00:38:22,226 ACTIVITY LEVEL AFTER LPS 934 00:38:22,226 --> 00:38:30,468 INJECTION INTO THE MICE 6 HOURS 935 00:38:30,468 --> 00:38:34,839 AFTER INFECTION. 936 00:38:34,839 --> 00:38:40,278 DEMON TRAITING LPS CAN REDUCE 937 00:38:40,278 --> 00:38:41,245 SMAD 3 PATHWAY, SO WHAT IS THE 938 00:38:41,245 --> 00:38:46,617 ROLE OF THIS PATHWAY IN TERMS OF 939 00:38:46,617 --> 00:38:48,353 INFLAMMATION AND SURVIVAL, IF 940 00:38:48,353 --> 00:38:50,421 YOU INDUCE SEPSIS IN THIS CREE 941 00:38:50,421 --> 00:38:52,390 MY, HAVE YOU DECREASED SURVIVAL, 942 00:38:52,390 --> 00:38:57,095 AND THIS IS LINKED TO INCREASED 943 00:38:57,095 --> 00:38:57,595 LEVELS OF INFLAMMATION. 944 00:38:57,595 --> 00:38:58,062 CERTAINLY--CERTAINLY 945 00:38:58,062 --> 00:39:08,606 SPECIALENTIAL LOAMACYY -I WELL, 946 00:39:15,613 --> 00:39:19,617 6, YOU HAVE SO, A QUESTION THAT 947 00:39:19,617 --> 00:39:21,152 WE WANTED TO UNDERSTAND IS OKAY, 948 00:39:21,152 --> 00:39:22,820 IN IS TRUE IN THE CONTEXT OF 949 00:39:22,820 --> 00:39:25,723 LPS, WHICH IS A BACTERIAL LIGAND 950 00:39:25,723 --> 00:39:30,661 AND A TNF LIGAND BUT WHAT ABOUT 951 00:39:30,661 --> 00:39:31,796 OTHER TOLL-LIKE RECEPTORS THAT 952 00:39:31,796 --> 00:39:33,531 ARE ABLE TO MEDIATE THIS EFFECT, 953 00:39:33,531 --> 00:39:35,600 WHAT ABOUT THE LIAISON GAPPEDS, 954 00:39:35,600 --> 00:39:36,968 AND ALSO WHAT ABOUT ENDOSTUDIES 955 00:39:36,968 --> 00:39:38,035 OF MULTIPLE ENDOCRINAL RECEPTORS 956 00:39:38,035 --> 00:39:41,739 SO TO TRY TO ANSWER THOSE 957 00:39:41,739 --> 00:39:43,808 QUESTIONS, WE FIRST USED THE 958 00:39:43,808 --> 00:39:44,642 EPROTEIN FROM [INDISCERNIBLE] 959 00:39:44,642 --> 00:39:45,943 THAT HAS BEEN DEMONSTRATED HERE, 960 00:39:45,943 --> 00:39:50,548 AND BE ABLE TO INFLAMMATION, AND 961 00:39:50,548 --> 00:39:53,251 WE ALSO USE TLR7 LIGAND WHICH IS 962 00:39:53,251 --> 00:39:56,587 MORE IN THE CONTEXT OF 963 00:39:56,587 --> 00:39:56,921 AUTOIMMUNITY. 964 00:39:56,921 --> 00:40:00,057 AND SO, SHOULD SHOULD GIVE 965 00:40:00,057 --> 00:40:01,359 PROTEIN TO MACROPHAGES INVITRO, 966 00:40:01,359 --> 00:40:03,795 YOU CAN INCREASE THE EXPRESSION 967 00:40:03,795 --> 00:40:10,735 OF ACTIN A AND YOU CAN ALSO 968 00:40:10,735 --> 00:40:13,571 INCREASE THE PHOSPHOROUS 3 969 00:40:13,571 --> 00:40:14,439 REGULATION. 970 00:40:14,439 --> 00:40:16,240 THE SAME THING HAPPENS IN 971 00:40:16,240 --> 00:40:18,309 RESPONSE TO, YOU HAVE INCREASED 972 00:40:18,309 --> 00:40:20,311 IN ACTIVITY A, AND HAVE YOU 973 00:40:20,311 --> 00:40:22,013 INCREASED SMAD 3 PHOSPHORYLATION 974 00:40:22,013 --> 00:40:26,083 WHICH IS COMPLETELY DEPENDENT ON 975 00:40:26,083 --> 00:40:28,186 AND DEMONSTRATING THOSE 2 976 00:40:28,186 --> 00:40:30,288 LIGANDS CAN INDUCE ACTIVE A AND 977 00:40:30,288 --> 00:40:32,457 PHOSPHORYLATION AND THAT IT'S A 978 00:40:32,457 --> 00:40:35,059 GENERAL PHENOMENON THAT SEEMS TO 979 00:40:35,059 --> 00:40:35,626 BE OCCURRING. 980 00:40:35,626 --> 00:40:37,695 SO WE DID QUITE A BIT OF WORK ON 981 00:40:37,695 --> 00:40:39,330 THE PROTEIN MODEL IN VIVO AND 982 00:40:39,330 --> 00:40:39,664 INVITRO. 983 00:40:39,664 --> 00:40:44,101 I WILL NOT SHOW THOSE DATA TODAY 984 00:40:44,101 --> 00:40:46,737 AND I WILL FOCUS MORE ON THE 985 00:40:46,737 --> 00:40:49,307 IMMUNITY PART AND THE 986 00:40:49,307 --> 00:40:50,208 [INDISCERNIBLE] AND SO CAN THE 987 00:40:50,208 --> 00:40:51,709 WHAD CAN BE APPLIED ON THE BACK 988 00:40:51,709 --> 00:40:53,678 SKIN OF THE MICE FOR 6 989 00:40:53,678 --> 00:40:58,749 CONSECUTIVE DAYS AND THAT CAN 990 00:40:58,749 --> 00:40:59,350 INDUCE [INDISCERNIBLE] DISEASE 991 00:40:59,350 --> 00:41:01,552 EMPLOY SO WE DID THE SAME THING, 992 00:41:01,552 --> 00:41:03,221 WE IMAGED FOR INFLAMMATION, WE 993 00:41:03,221 --> 00:41:04,655 DID IT FOR 6 HOURS AND WE USE IT 994 00:41:04,655 --> 00:41:08,526 FOR THE ACTIVATION OF THIS 995 00:41:08,526 --> 00:41:11,128 [INDISCERNIBLE] PATHWAY. 996 00:41:11,128 --> 00:41:12,196 SO IF YOU APPLY [INDISCERNIBLE] 997 00:41:12,196 --> 00:41:13,865 ON THE MICE AFTER THE FIRST 998 00:41:13,865 --> 00:41:19,103 APPLICATION, HAVE YOU INCRACIED 999 00:41:19,103 --> 00:41:20,805 ACTIVE EXPRESSION, AND YOU HAVE 1000 00:41:20,805 --> 00:41:22,673 INCREASED LEVEL OF SMAD 3 1001 00:41:22,673 --> 00:41:24,242 PHOSPHORYLATION AS CAN YOU 1002 00:41:24,242 --> 00:41:26,177 APPRECIATE HERE, DEMONSTRATE 1003 00:41:26,177 --> 00:41:28,012 THANKSGIVING PATHWAY IS INDUCED. 1004 00:41:28,012 --> 00:41:31,916 SO NOW WHAT IS THE FUNCTION OF 1005 00:41:31,916 --> 00:41:33,317 THIS PATHWAY, IF WE USE 1006 00:41:33,317 --> 00:41:34,118 [INDISCERNIBLE] CREE MICE AND 1007 00:41:34,118 --> 00:41:36,654 APPLY ON THE BACK OF THE MICE, 1008 00:41:36,654 --> 00:41:39,524 SO, THOSE MICE HAVE A HIGHER 1009 00:41:39,524 --> 00:41:40,625 [INDISCERNIBLE] WHICH MEANS THEY 1010 00:41:40,625 --> 00:41:41,893 ARE MORE SICK, AND WE COULD 1011 00:41:41,893 --> 00:41:44,228 OBSERVE THAT WE HAVE MORE 1012 00:41:44,228 --> 00:41:46,364 MACROPHAGES IN THE SKIN AND 1013 00:41:46,364 --> 00:41:49,133 THOSE MACROPHAGES PRODUCE MORE 1014 00:41:49,133 --> 00:41:50,835 IL6 SO THEY'RE MORE 1015 00:41:50,835 --> 00:41:51,302 INFLAMMATORY. 1016 00:41:51,302 --> 00:41:52,670 AND NOW THEY BECOME MORE 1017 00:41:52,670 --> 00:41:55,039 INFLAMMATORY, THEY WILL DRIVE 1018 00:41:55,039 --> 00:41:58,075 THE GAMMA T-CELLS AND THEY WILL 1019 00:41:58,075 --> 00:42:00,645 STOP PRODUCING IL15 1020 00:42:00,645 --> 00:42:01,445 INTERFERON-GAMMA ANDILE L22. 1021 00:42:01,445 --> 00:42:02,613 AND DEMON TRAIT THANKSGIVING 1022 00:42:02,613 --> 00:42:05,249 ACTIVE A AS THE PATHWAY CAN 1023 00:42:05,249 --> 00:42:08,219 CONTROL THE MACROPHAGES BEHAVIOR 1024 00:42:08,219 --> 00:42:10,321 THAT CAN BE PROINFLAMMATORY AND 1025 00:42:10,321 --> 00:42:16,961 CONTROL THE ACTIVATION OF THE 1026 00:42:16,961 --> 00:42:21,666 GAMMA T-CELLS DURING SORTING 1027 00:42:21,666 --> 00:42:23,000 SIGNALS AYA SIS. 1028 00:42:23,000 --> 00:42:26,137 AND WEEE INYEBED ACTIVEIN A INTO 1029 00:42:26,137 --> 00:42:28,072 THE INTRA DERMALLY INTO THE SKIN 1030 00:42:28,072 --> 00:42:33,177 OF THE MICE AND SO IF YOU INJECT 1031 00:42:33,177 --> 00:42:36,614 ACTIVEIN INTO THE MICE, THIS IS 1032 00:42:36,614 --> 00:42:41,052 DEPENDENT ON SMAD 3, SO THE 1033 00:42:41,052 --> 00:42:43,054 EFFECTIVE IT IN THERE IS 1034 00:42:43,054 --> 00:42:43,421 [INDISCERNIBLE]. 1035 00:42:43,421 --> 00:42:45,323 AND IN THOSE TREATED MICE WE 1036 00:42:45,323 --> 00:42:47,992 OBSERVED A DECREASED LEVEL OF 1037 00:42:47,992 --> 00:42:51,062 MACROPHAGES AND THIS IS 1038 00:42:51,062 --> 00:42:52,563 DEPENDENT ON SMAD 3, AND OF 1039 00:42:52,563 --> 00:42:54,265 COURSE WE HAVE DECREASE 1040 00:42:54,265 --> 00:42:55,499 EXPRESSION OF IL17 AND GAMMA AND 1041 00:42:55,499 --> 00:42:57,535 THIS IS AGAIN COMPLETELY 1042 00:42:57,535 --> 00:43:00,905 REVERTED TO MORE LEVELS IN SMAD 1043 00:43:00,905 --> 00:43:03,207 3 KNOCK OUT DEFESHT MICE. 1044 00:43:03,207 --> 00:43:05,543 SO DEMONSTRATING THIS ACTIVE A 1045 00:43:05,543 --> 00:43:09,747 PATHWAY CAN CONTROL INDUCED THE 1046 00:43:09,747 --> 00:43:12,717 [INDISCERNIBLE] AND BE A BREAK 1047 00:43:12,717 --> 00:43:14,018 FOR INPOLICEMANNATION AND IPT 1048 00:43:14,018 --> 00:43:14,852 GREATERUNITY AND ACCEPTIS. 1049 00:43:14,852 --> 00:43:21,392 SO FINALLY IS THIS PATHWAY 1050 00:43:21,392 --> 00:43:23,260 ACTIVE IN HUMAN IN SO WE HOPE TO 1051 00:43:23,260 --> 00:43:25,696 HAVE MORE DATA ON THIS AS WELL. 1052 00:43:25,696 --> 00:43:30,668 SO WE CAME BACK TO THE DATA FROM 1053 00:43:30,668 --> 00:43:32,069 THE FIRST PROJECT AND WE 1054 00:43:32,069 --> 00:43:33,137 OBSERVED THIS PATHWAY TO BE 1055 00:43:33,137 --> 00:43:36,140 ACTIVE IF PATIENTS THAT HAVE 1056 00:43:36,140 --> 00:43:37,508 SEPSIS, DEVELOP SEPSIS, 1057 00:43:37,508 --> 00:43:40,177 INCREASED LEVEL OF ACTIVITY A, 1058 00:43:40,177 --> 00:43:43,648 3, RECEPTOR IN STAT 5, IN 1059 00:43:43,648 --> 00:43:46,717 MACROPHAGES AND IN COVID-19, YOU 1060 00:43:46,717 --> 00:43:49,086 ALSO HAVE INCREASED EXPRESSION 1061 00:43:49,086 --> 00:43:52,356 OF THOSE YEENS IN COVID-19 1062 00:43:52,356 --> 00:43:52,623 PATIENTS. 1063 00:43:52,623 --> 00:43:53,324 DEMONSTRATING THIS ACTIVITY OF 1064 00:43:53,324 --> 00:43:56,661 THE PATHWAY COULD ALSO BE ACTIVE 1065 00:43:56,661 --> 00:44:00,297 IN PATIENT'S THAT DEVELOP SEPSIS 1066 00:44:00,297 --> 00:44:01,132 AND COVID-19. 1067 00:44:01,132 --> 00:44:03,000 SO TO CONCLUDE THIS PART OF THE 1068 00:44:03,000 --> 00:44:08,439 TALK, WHAT I SHOWED YOU IF 1069 00:44:08,439 --> 00:44:09,507 THAT'S LPS, PROTEIN, AND 1070 00:44:09,507 --> 00:44:11,042 [INDISCERNIBLE], SIGNALS WITH 1071 00:44:11,042 --> 00:44:12,777 THEIR TLRs AND CAN INDUCE STAT 1072 00:44:12,777 --> 00:44:13,944 5 ACTIVATION, SO I DID NOT SHOW 1073 00:44:13,944 --> 00:44:15,846 IT TO YOU FOR A PROTEIN, BUT 1074 00:44:15,846 --> 00:44:17,348 BASICALLY THEY HAVE THE SAME 1075 00:44:17,348 --> 00:44:20,217 THING IN PATHWAY THROUGH MYD88 1076 00:44:20,217 --> 00:44:21,719 STAT PATHWAY AND AT THAT TIME 5, 1077 00:44:21,719 --> 00:44:24,588 AND THEN STAT 5 CAN BIND TO THE 1078 00:44:24,588 --> 00:44:28,225 PROMOTER OF ACTIVEIN A, CAN 1079 00:44:28,225 --> 00:44:29,794 PRODUCE THE PRODUCTION OF 1080 00:44:29,794 --> 00:44:31,562 ACTIVIN A, BUT WE BELIEVE IT'S 1081 00:44:31,562 --> 00:44:34,732 HAPPENING IN OTHER CONTEXT, IT 1082 00:44:34,732 --> 00:44:35,766 CAN INDUCE PHOSPHORYLATION, SMAD 1083 00:44:35,766 --> 00:44:40,071 3 CAN BIND TO SELF-PROMOTERS AND 1084 00:44:40,071 --> 00:44:41,839 REGULATE INFLAMMATION WITH 1085 00:44:41,839 --> 00:44:47,578 METABOLISM DURING SEPSIS, SUCH 1086 00:44:47,578 --> 00:44:47,945 AS IN COVID-19. 1087 00:44:47,945 --> 00:44:49,146 SO WHAT ARE THE FUTURE 1088 00:44:49,146 --> 00:44:51,282 DIRECTIONS OF THIS WORK? 1089 00:44:51,282 --> 00:44:53,317 SO 1 OF THE DIRECTION WHICH IS 1090 00:44:53,317 --> 00:44:55,186 RELATED TO THE FIRST PROJECT IS 1091 00:44:55,186 --> 00:44:58,823 THAT WE STUDIED THE ROLE OF 1092 00:44:58,823 --> 00:45:00,157 MACROPHAGES IN THE ROLE OF 1093 00:45:00,157 --> 00:45:01,492 RESPONSE TO TGF BETA AND 1094 00:45:01,492 --> 00:45:03,027 INFECTION ANDS VERY HIGH LEVELS 1095 00:45:03,027 --> 00:45:03,761 OF INFLAMMATION BUT WHAT 1096 00:45:03,761 --> 00:45:05,362 HAPPENING IF YOU HAVE HIGH 1097 00:45:05,362 --> 00:45:07,932 LEVELS OF TGF BETA AND LOW GRADE 1098 00:45:07,932 --> 00:45:14,872 OF INFLAMMATION, ESPECIALLY THE 1099 00:45:14,872 --> 00:45:16,407 RULE OF TUMOR MICROENVIRONMENT. 1100 00:45:16,407 --> 00:45:19,143 ONE POINT WHICH I THINK IS 1101 00:45:19,143 --> 00:45:21,846 EXTREMELY IMPORTANT, COULD WE 1102 00:45:21,846 --> 00:45:25,549 TARGET THE TGF BETAACIS DURING 1103 00:45:25,549 --> 00:45:26,717 SEPSIS IN HUMANS, SO OF COURSE, 1104 00:45:26,717 --> 00:45:28,619 THERE IS SOME WORK REQUIRED THAT 1105 00:45:28,619 --> 00:45:33,124 WE--THAT WOULD REQUIRE TO 1106 00:45:33,124 --> 00:45:34,792 DEVELOP SOME ANTIBODIES AND 1107 00:45:34,792 --> 00:45:39,497 INHIBITORS, BUT WE HAVE ALL 1108 00:45:39,497 --> 00:45:40,865 TOOLS AVAILABLE AND INHIBITORS 1109 00:45:40,865 --> 00:45:41,766 AND ACTIVATION THAT ARE 1110 00:45:41,766 --> 00:45:50,107 AVAILABLE ALREADY AND THERE IS 1111 00:45:50,107 --> 00:45:52,943 NOW AND DRIVE THIS, WE WILL HEAR 1112 00:45:52,943 --> 00:46:01,418 THE GOOD COAGULATION, AND IT'S 1113 00:46:01,418 --> 00:46:02,686 BEEN TARGETED WE WOULD LIKE TO 1114 00:46:02,686 --> 00:46:04,288 COLLABORATE AND BE MORE THAN 1115 00:46:04,288 --> 00:46:04,688 HAPPY. 1116 00:46:04,688 --> 00:46:05,990 AND THEN IA LITTLE BIT ABOUT THE 1117 00:46:05,990 --> 00:46:07,792 PROGECS I SHOW YOU DURING 1118 00:46:07,792 --> 00:46:13,864 SEPSIS, ACTIN A IS A GOOD GUY 1119 00:46:13,864 --> 00:46:15,232 BUT WE WANT TO TRY TO UNDERSTAND 1120 00:46:15,232 --> 00:46:17,168 WHAT CONTEXT THIS IS MORE 1121 00:46:17,168 --> 00:46:18,502 IMPORTANT THAN TGF BETA, WE 1122 00:46:18,502 --> 00:46:20,204 BELIEVE IN THE EXAMPLE OF THE 1123 00:46:20,204 --> 00:46:21,438 TUMOR MICROENVIRONMENT, IT COULD 1124 00:46:21,438 --> 00:46:23,140 BE MORE IMPORTANT THAN ACTIN A 1125 00:46:23,140 --> 00:46:24,508 BUT IT DEPENDS ON ALSO THE 1126 00:46:24,508 --> 00:46:25,910 ACTIVATION OF THOSE PROTEINS AND 1127 00:46:25,910 --> 00:46:28,345 MANY DIFFERENT FACTORS AND HOW 1128 00:46:28,345 --> 00:46:31,582 ARE THEY DIFREPRESENTIALLY 1129 00:46:31,582 --> 00:46:33,150 REGULATE, SPECIALLY WE FOCUS ON 1130 00:46:33,150 --> 00:46:34,985 SMAD 3 ACTIVATE BOOED I BOTH BUT 1131 00:46:34,985 --> 00:46:36,987 THEY HAVE DIFFERENT SIGNALING 1132 00:46:36,987 --> 00:46:40,891 PATHWAY, LIKE mTOR AND 1133 00:46:40,891 --> 00:46:43,727 SIGNALING FOR TGF BETA SO WHICH 1134 00:46:43,727 --> 00:46:45,563 PATHWAY IS MORE IMPORTANT AND 1135 00:46:45,563 --> 00:46:48,232 FINALLY WE COULD USE ACT VIN A 1136 00:46:48,232 --> 00:46:50,634 TO FIGHT INFECTIONS IN HUMAN AND 1137 00:46:50,634 --> 00:46:52,369 TO DECREASE INFLAMMATION AND 1138 00:46:52,369 --> 00:46:57,007 INDUCE A RETURN TO HOMEOSTASIS. 1139 00:46:57,007 --> 00:46:58,809 SO THE LAST SLIDE BUT OF COURSE 1140 00:46:58,809 --> 00:46:59,577 ALSO THE MOST IMPORTANT. 1141 00:46:59,577 --> 00:47:01,111 SO I WILL LIKE TO THANK 1142 00:47:01,111 --> 00:47:04,215 EVERYBODY WHO HELPED ME FOR THE 1143 00:47:04,215 --> 00:47:09,520 STUDIES, MY MENTOR DR. CHEN, AN 1144 00:47:09,520 --> 00:47:10,721 AMAZING MENTOR THESE PAST 5 1145 00:47:10,721 --> 00:47:12,056 YEARS, AS WELL AS EVERYBODY IN 1146 00:47:12,056 --> 00:47:14,458 THE LAB WHO IN THE PICTURE WHO 1147 00:47:14,458 --> 00:47:17,795 HELPED ME PERFORM SO MANY SPRPTS 1148 00:47:17,795 --> 00:47:20,497 AS WELL AS [INDISCERNIBLE], I 1149 00:47:20,497 --> 00:47:25,636 WOULD ALSO LIKE TO THANK ALL OUR 1150 00:47:25,636 --> 00:47:26,737 COLLABORATORS ESPECIALLY INSIDE 1151 00:47:26,737 --> 00:47:32,743 NIH, FOR THE RNASEQ ANALYSIS AND 1152 00:47:32,743 --> 00:47:34,144 DR. CHEN YRK AO, AND 1153 00:47:34,144 --> 00:47:36,981 [INDISCERNIBLE] FOR THE HELP IN 1154 00:47:36,981 --> 00:47:43,687 ATTACK SEQ AND CHIP ANALYSIS, 1155 00:47:43,687 --> 00:47:48,158 DR. LARRION AND DR. BEWLY, AND 1156 00:47:48,158 --> 00:47:50,327 DR. SU FOR PROVIDING THE ACR1 B 1157 00:47:50,327 --> 00:47:51,962 FLUX MICE AND I WOULD LIKE TO 1158 00:47:51,962 --> 00:47:52,997 THANK YOU FOR YOUR ATTENTION AND 1159 00:47:52,997 --> 00:47:55,132 IF YOU HAVE ANY QUESTIONS, THE 1160 00:47:55,132 --> 00:48:02,306 FLOOR IS YOURS. 1161 00:48:02,306 --> 00:48:02,806 [ APPLAUSE ] 1162 00:48:02,806 --> 00:48:04,241 >> THANK YOU EMPLOY WE HAVE 1163 00:48:04,241 --> 00:48:05,976 QUESTIONS COMING ALREADY SO I 1164 00:48:05,976 --> 00:48:07,478 WILL HOLD MINE BACK. 1165 00:48:07,478 --> 00:48:08,779 >> INTERESTING TALK, I HAVE A 1166 00:48:08,779 --> 00:48:11,081 FEW QUESTIONS REGARDING THE 1167 00:48:11,081 --> 00:48:12,917 FIRST PART EMPLOY ANDIME SO CURE 1168 00:48:12,917 --> 00:48:15,886 QUOWS WHAT IS GOING TO HAPPEN IF 1169 00:48:15,886 --> 00:48:18,856 YOU TAKE THE MACROPHAGE THAT THE 1170 00:48:18,856 --> 00:48:20,824 TGF BETA DEFESHT RECEPTOR AND 1171 00:48:20,824 --> 00:48:22,559 YOU STIMULATE THEM WITHOUT THE 1172 00:48:22,559 --> 00:48:24,762 [INDISCERNIBLE], THEY WILL 1173 00:48:24,762 --> 00:48:26,297 REGULATE GLYCOLYSIS OR WHAT DO 1174 00:48:26,297 --> 00:48:26,530 THEY DO? 1175 00:48:26,530 --> 00:48:27,965 >> YES, SO WE DIDN'T GO THAT 1176 00:48:27,965 --> 00:48:31,302 MUCH FURTHER ON THAT, BUT IT 1177 00:48:31,302 --> 00:48:32,102 LOOKS LIKE, YES. 1178 00:48:32,102 --> 00:48:34,772 AND BAGGED ON THAT, MY QUESTION 1179 00:48:34,772 --> 00:48:37,641 IS WHAT DO THOSE MACROPHAGE DO 1180 00:48:37,641 --> 00:48:39,043 BECAUSE YOU REGULATE DPLI COLSIS 1181 00:48:39,043 --> 00:48:40,878 AND THEY WILL PRODUCE 1182 00:48:40,878 --> 00:48:43,580 INTERLEUKIN 1 BETA BY ACTIVATING 1183 00:48:43,580 --> 00:48:45,082 INFLAMMASOME OR WHAT DO THEY 1184 00:48:45,082 --> 00:48:45,516 PROTIEWS? 1185 00:48:45,516 --> 00:48:48,786 >> WE DIDN'T SEE THAT MUCH IL1 1186 00:48:48,786 --> 00:48:52,456 BETA CHANGES ESPECIALLY IN THE 1187 00:48:52,456 --> 00:48:54,191 INPOLICEMANNASOME ACTIVATION, 1188 00:48:54,191 --> 00:48:55,225 AND PROINFLAMMATORYITE O KINES, 1189 00:48:55,225 --> 00:48:56,593 AND HAVE YOU CHECKED BY CHANCE 1190 00:48:56,593 --> 00:48:58,896 WHEN YOU KNOCK DOWN, I DON'T 1191 00:48:58,896 --> 00:49:04,568 KNOW IF YOU DO NOT TGFPAYS BETA 1192 00:49:04,568 --> 00:49:06,036 TO THE MACROPHAGE OR SOMETHING, 1193 00:49:06,036 --> 00:49:08,172 IN THE FAIG O SITTIC CAPACITY OF 1194 00:49:08,172 --> 00:49:09,573 THE CHANGES WHEN YOUACIVATE THEM 1195 00:49:09,573 --> 00:49:13,944 IN THE PRESENCE OF THE TGF BETA 1196 00:49:13,944 --> 00:49:14,745 OR NOT BECAUSE YOU DON'T KNOW 1197 00:49:14,745 --> 00:49:17,314 WHEN YOU PUT THE TGF BETA INTO 1198 00:49:17,314 --> 00:49:20,351 MACROPHAGES THEY ARE BECOMING 1199 00:49:20,351 --> 00:49:21,618 mTOR MACROPHAGES NSO TO ANSWER, 1200 00:49:21,618 --> 00:49:22,820 THE FIRST THING WE DID NOT TRY 1201 00:49:22,820 --> 00:49:24,989 TO DO THAT BUT IT'S A VERY GOOD 1202 00:49:24,989 --> 00:49:25,556 POINT. 1203 00:49:25,556 --> 00:49:28,559 WE DID NOT LOOK AT PHAGOCYTOSIS, 1204 00:49:28,559 --> 00:49:30,427 SO WE CAN TELL YOU THERE ARE NO 1205 00:49:30,427 --> 00:49:31,795 DIFFERENCE IN THE LEVEL IN THE 1206 00:49:31,795 --> 00:49:34,498 BLOOD OF THE MICE BUT LOOKING AT 1207 00:49:34,498 --> 00:49:36,934 PHAGOCYTOSIS WOULD BE VERY 1208 00:49:36,934 --> 00:49:37,201 IMPORTANT. 1209 00:49:37,201 --> 00:49:39,136 I FORGET WHAT I WANTED TO SAY 1210 00:49:39,136 --> 00:49:45,275 FOR A SECOND PART. 1211 00:49:45,275 --> 00:49:50,180 WHAT WAS THE END OF THE 1212 00:49:50,180 --> 00:49:50,447 QUESTION. 1213 00:49:50,447 --> 00:49:53,183 >> THE FAIG O SITTIC CAPACITY 1214 00:49:53,183 --> 00:49:54,952 DIFFERENT BETWEEN THE USE 1215 00:49:54,952 --> 00:49:56,053 STIMULATOR [INDISCERNIBLE] 1216 00:49:56,053 --> 00:49:56,453 TGF BETA? 1217 00:49:56,453 --> 00:49:58,389 >> YEAH, SO WE DID NOT LOOK AT 1218 00:49:58,389 --> 00:50:03,027 THAT, WE KNOW THAT IN CONTEXT OF 1219 00:50:03,027 --> 00:50:04,294 THE SIGNIFYITOSEIS THAT WE DID 1220 00:50:04,294 --> 00:50:08,766 BUT NOT IN CONTEXT, BUT IN VIVO 1221 00:50:08,766 --> 00:50:11,735 THERE'S NO DIFFERENCE IN LEVELS 1222 00:50:11,735 --> 00:50:12,936 OF INFECTION OF THOSE INTO THE 1223 00:50:12,936 --> 00:50:14,271 BLOOD OF THE MICE. 1224 00:50:14,271 --> 00:50:16,507 IN, OKAY, THANK YOU NYOU'RE 1225 00:50:16,507 --> 00:50:16,740 WELCOME. 1226 00:50:16,740 --> 00:50:17,941 NI HAVE A QUESTION ABOUT THE 1227 00:50:17,941 --> 00:50:19,343 FITTER PART OFIOURE TALK AS 1228 00:50:19,343 --> 00:50:24,448 WELL, VERY NICE TALK, SO, 1229 00:50:24,448 --> 00:50:26,050 COAGULATION, FACTOR 13 IS THE 1230 00:50:26,050 --> 00:50:28,485 END STAGE WOULD ACTUALLY TABLIZE 1231 00:50:28,485 --> 00:50:30,354 THE CLOT, AND LPS I'M SURE 1232 00:50:30,354 --> 00:50:32,689 INDUCES CLOTTING FACTORS FOR 1233 00:50:32,689 --> 00:50:33,957 EXAMPLE, INDUCES ALL RECOLLECT 1234 00:50:33,957 --> 00:50:35,359 FACTORS AND COMPLEMENT 1235 00:50:35,359 --> 00:50:41,398 ACTIVATION, ET CETERA, DO YOU 1236 00:50:41,398 --> 00:50:43,400 THINK TGF BETAS THOSE LESMS, 7, 1237 00:50:43,400 --> 00:50:45,469 8, 9, WHATEVER YOU WANT TO CALL 1238 00:50:45,469 --> 00:50:47,104 THEM, THE BLOOD CLOTTING IS GOOD 1239 00:50:47,104 --> 00:50:49,239 AND HOW IS THIS STACK FACTOR THE 1240 00:50:49,239 --> 00:50:50,874 MOST IMPORTANT AS OPPOSE TO 1241 00:50:50,874 --> 00:50:54,011 CHANGING LEVELS INCREASES OF 1242 00:50:54,011 --> 00:50:55,846 OTHER BLOOD CLOTTING LEVELS 1243 00:50:55,846 --> 00:50:56,480 AFTER SEPSIS. 1244 00:50:56,480 --> 00:50:58,982 >> THAT'S A VERY GOOD QUESTION 1245 00:50:58,982 --> 00:50:59,817 ACTUALLY, UNFORTUNATELY WE WERE 1246 00:50:59,817 --> 00:51:03,087 NOT ABLE TO GO FURTHER INTO THE 1247 00:51:03,087 --> 00:51:05,022 COAGULATION PART SO WE WOULD 1248 00:51:05,022 --> 00:51:07,191 HAVE LOVED TO HAVE THOSE MYSELF 1249 00:51:07,191 --> 00:51:09,393 AND WITH CREE AND PROVE THAT F13 1250 00:51:09,393 --> 00:51:10,461 IS THE MOST IMPORTANT BUT WE 1251 00:51:10,461 --> 00:51:13,363 WERE NOT ABLE TO GO THAT FAR SO 1252 00:51:13,363 --> 00:51:15,432 YEAH, THE THINK OF THAT BECAUSE 1253 00:51:15,432 --> 00:51:16,733 WE INJECTED TGF BETA AND BLOCKED 1254 00:51:16,733 --> 00:51:18,235 TGF BETA WE BELIEVE THAT THERE 1255 00:51:18,235 --> 00:51:22,406 MIGHT BE OTHER CELLS AFFECTED BY 1256 00:51:22,406 --> 00:51:25,476 TGF BETA LIKE [INDISCERNIBLE] OR 1257 00:51:25,476 --> 00:51:27,911 OTHERS THAT PLAY A ROLE IN 1258 00:51:27,911 --> 00:51:28,612 COAGULATION NDID YOU SEE 1259 00:51:28,612 --> 00:51:30,147 CLOTTING IN THE ANIMALS? 1260 00:51:30,147 --> 00:51:32,249 WHERE DOES IT OCCUR? 1261 00:51:32,249 --> 00:51:34,785 IN SEPSIS WHERE DO YOU SEE A LOT 1262 00:51:34,785 --> 00:51:34,985 IN. 1263 00:51:34,985 --> 00:51:37,688 >> IN THE BLOOD IF YOU TAKE THE 1264 00:51:37,688 --> 00:51:39,323 BLOOD, THE COAGULATION IS MUCH 1265 00:51:39,323 --> 00:51:40,023 FASTER. 1266 00:51:40,023 --> 00:51:46,296 >> OKAY, THANK YOU NYOU'RE 1267 00:51:46,296 --> 00:51:46,597 WELCOME. 1268 00:51:46,597 --> 00:51:51,268 >> WE CAN LET TRAINEES GO FIRST, 1269 00:51:51,268 --> 00:51:51,468 RIGHT? 1270 00:51:51,468 --> 00:51:56,106 , TRAINEES FIRST. 1271 00:51:56,106 --> 00:51:59,843 >> SO, WHEN YOU HAVE CYTOKINE 1272 00:51:59,843 --> 00:52:01,245 STORM TOXIC SHOCK CONNECK WIDE 1273 00:52:01,245 --> 00:52:06,683 SEPSIS FOR EXAMPLE OR OTHER 1274 00:52:06,683 --> 00:52:12,156 SITUATIONS THERE'S OFTEN IL6 1275 00:52:12,156 --> 00:52:13,323 INVOLVED AND TREATMENT WITH 1276 00:52:13,323 --> 00:52:15,058 [INDISCERNIBLE] WHICH IS AN IL6 1277 00:52:15,058 --> 00:52:17,694 INHIBITOR CAN BE EFFECTIVE AND 1278 00:52:17,694 --> 00:52:21,098 WE KNOW THAT TGF BETA CAN 1279 00:52:21,098 --> 00:52:23,433 COLLABORATE WITH IL6 TO INDUCE 1280 00:52:23,433 --> 00:52:26,937 TH15 AND ILPANE PRODUCTION, SO, 1281 00:52:26,937 --> 00:52:31,341 HOW DOES ALL THAT FIT INTO YOUR 1282 00:52:31,341 --> 00:52:33,076 SEPSIS MODEL WHERE I DON'T 1283 00:52:33,076 --> 00:52:38,015 RECALL YOUR MENTIONING EITHER 1284 00:52:38,015 --> 00:52:42,486 IL6 OR IL15 EXCEPT LATER IN THE 1285 00:52:42,486 --> 00:52:45,556 CONTEXT OF PSORIASIS, YOU 1286 00:52:45,556 --> 00:52:45,956 MENTIONED IL17. 1287 00:52:45,956 --> 00:52:47,324 SMRKS GOOD QUESTION, SO THE 1288 00:52:47,324 --> 00:52:49,493 ACTUALLY THE LEVELS OF HAVE NOT 1289 00:52:49,493 --> 00:52:50,627 CHANGED IN MACROPHAGES OR IN THE 1290 00:52:50,627 --> 00:52:51,995 BLOOD OF THOSE MICE. 1291 00:52:51,995 --> 00:52:53,330 THEY DON'T SEEM TO BE AFFECTED, 1292 00:52:53,330 --> 00:52:56,099 WE COULD NOT SEE ANY TH17 1293 00:52:56,099 --> 00:52:59,269 GENERATION BUT IT'S PRETTY EARLY 1294 00:52:59,269 --> 00:52:59,603 ON. 1295 00:52:59,603 --> 00:53:01,772 AND THEY'RE LIKE 18 HOURS AND 1296 00:53:01,772 --> 00:53:03,674 THEN, LIKE EVERYBODY THAT HAS TO 1297 00:53:03,674 --> 00:53:05,175 DIE, DIES BY 48 HOURS, SO IT'S 1298 00:53:05,175 --> 00:53:08,178 PROBABLY STILL A LITTLE BIT 1299 00:53:08,178 --> 00:53:11,415 EARLY TO SEE DRAMATIC TH17 1300 00:53:11,415 --> 00:53:12,416 RESPONSE, SO PROBABLY WHY WE 1301 00:53:12,416 --> 00:53:14,318 DIDN'T SEE IT NOW IN LONGER 1302 00:53:14,318 --> 00:53:15,485 INFECS, YEAH, PROBABLY I THINK 1303 00:53:15,485 --> 00:53:18,622 IT COULD PLAY A ROLE, YEAH, 1304 00:53:18,622 --> 00:53:26,296 DEFINITELY, YEAH. 1305 00:53:26,296 --> 00:53:27,564 >> OKAY, THANK YOU. 1306 00:53:27,564 --> 00:53:28,632 >> THANK YOU, THAT WAS A NICE 1307 00:53:28,632 --> 00:53:28,865 TALK. 1308 00:53:28,865 --> 00:53:31,068 I HAVE A QUESTION ABOUT YOUR 1309 00:53:31,068 --> 00:53:32,369 FIRST PART MORE CONCEPT WISE 1310 00:53:32,369 --> 00:53:33,637 BECAUSE YOU MENTIONED ABOUT 1311 00:53:33,637 --> 00:53:35,706 SEPSIS IN THE COVID INFECTION, 1312 00:53:35,706 --> 00:53:40,043 DO YOU THINK IT'S LIKE HOW CAN I 1313 00:53:40,043 --> 00:53:42,779 SAY THE REAL--I DON'T FINISH 1314 00:53:42,779 --> 00:53:45,549 IT'S THE RIGHT WORD BUT REAL 1315 00:53:45,549 --> 00:53:49,653 SETTING OF TGF BETA AND THOSE 1316 00:53:49,653 --> 00:53:50,354 LIKE SIGNALING PATHWAYS INVOLVED 1317 00:53:50,354 --> 00:53:52,723 TO SAY IN A SETTING OF COVID-19 1318 00:53:52,723 --> 00:53:53,223 SITUATION? 1319 00:53:53,223 --> 00:53:55,392 DO YOU THINK THIS CYTOKINE STORM 1320 00:53:55,392 --> 00:53:59,296 MAY BE RELATED TO THOSE TGF BETA 1321 00:53:59,296 --> 00:54:00,697 SECRETION OR? 1322 00:54:00,697 --> 00:54:02,899 I JUST WANT TO SEE YOUR-- 1323 00:54:02,899 --> 00:54:05,002 >> SO YOU'RE ASKING WHETHER YOU 1324 00:54:05,002 --> 00:54:07,604 HAVE CYTOKINE STORM IN RESPONSE 1325 00:54:07,604 --> 00:54:09,039 TO TGF BETA? 1326 00:54:09,039 --> 00:54:11,308 >> RIGHT, SO IT LEADS TO THE 1327 00:54:11,308 --> 00:54:11,541 SEPSIS. 1328 00:54:11,541 --> 00:54:11,875 >> EXACTLY. 1329 00:54:11,875 --> 00:54:14,177 SO THE FIRST THING THAT WE 1330 00:54:14,177 --> 00:54:16,313 INVITRO IF YOU GET THE PROTEIN 1331 00:54:16,313 --> 00:54:17,347 FROM COVID-19, HAVE YOU THE SAME 1332 00:54:17,347 --> 00:54:19,082 THING, CAN YOU ACTIVATE TBLI 1333 00:54:19,082 --> 00:54:21,852 COLSIS BUT YOU CAN DECREASE 1334 00:54:21,852 --> 00:54:23,487 INFLAMMATION IN RESPONSE TO 1335 00:54:23,487 --> 00:54:24,621 TGF BETA SO THAT ANSWERED PART 1336 00:54:24,621 --> 00:54:25,589 OF YOUR QUESTION, THE 1337 00:54:25,589 --> 00:54:26,490 SECURITIZATION. THING, SO WE 1338 00:54:26,490 --> 00:54:29,626 LOOK AT INFLAMMATION AFTER ALL 1339 00:54:29,626 --> 00:54:31,628 THE WORKING ANS THERE IS NO MORE 1340 00:54:31,628 --> 00:54:32,829 SCIENCE OF INNAMES--NAMESSATION, 1341 00:54:32,829 --> 00:54:35,899 THE BRAIN, LUNGS, THE KIDNEY, 1342 00:54:35,899 --> 00:54:37,534 THE LIVER, WHEREVER WE LOOKED, 1343 00:54:37,534 --> 00:54:39,703 WE LOOKED AT CYTOKINES, THE 1344 00:54:39,703 --> 00:54:41,338 MACROPHAGES ARE STILL 1345 00:54:41,338 --> 00:54:42,773 ANTIINFLAMMATORY IN RESPONSE TO 1346 00:54:42,773 --> 00:54:44,041 TGF BETA SO WE REALLY BELIEVE 1347 00:54:44,041 --> 00:54:49,179 THAT IT'S NOT RELATED TO 1348 00:54:49,179 --> 00:54:52,049 CYTOKINE STORM BUT TISSUE 1349 00:54:52,049 --> 00:54:53,383 RELEVANCE AND DISCOAGULATION IS 1350 00:54:53,383 --> 00:54:55,085 DIFFERENT AND DRIVING THE MICE 1351 00:54:55,085 --> 00:54:57,387 BUT THE CYTOKINE STORM. 1352 00:54:57,387 --> 00:54:57,754 >> THANK YOU. 1353 00:54:57,754 --> 00:54:59,856 >> THANKS THAT WAS REALLY GREAT. 1354 00:54:59,856 --> 00:55:01,591 REALLY ENJOYED IT, SO I HAVE A 1355 00:55:01,591 --> 00:55:02,759 QUESTION ABOUT MRI COLSIS DATA 1356 00:55:02,759 --> 00:55:03,960 WHICH I THOUGHT WAS REALLY 1357 00:55:03,960 --> 00:55:05,996 CONVIPSING BUT I WANT TO THROW 1358 00:55:05,996 --> 00:55:08,665 OUT ANOTHER POSSIBILITY WHICH IS 1359 00:55:08,665 --> 00:55:09,700 JUST THAT METABOLISM IN GENERAL 1360 00:55:09,700 --> 00:55:12,202 IS NEEDED SO HAVE YOU TRIED 1361 00:55:12,202 --> 00:55:14,938 TREATING YOUR MICE FOR EXAMPLE 1362 00:55:14,938 --> 00:55:17,274 WITH A FLUTA MIN OLYMPIC SIS 1363 00:55:17,274 --> 00:55:20,877 INHIBITOR LIKE WITH DON OR 1364 00:55:20,877 --> 00:55:22,512 BPTSAND DO YOU GET THE SAME OR 1365 00:55:22,512 --> 00:55:25,716 DO YOU GET TBLI COLSIS. 1366 00:55:25,716 --> 00:55:26,783 >> GOOD QUESTION AGAIN, WE DID 1367 00:55:26,783 --> 00:55:28,985 NOT TRY TO BLOCK IT, WE TRY TO 1368 00:55:28,985 --> 00:55:30,887 BLOCK THE TCR SICK SEAL AND THEN 1369 00:55:30,887 --> 00:55:32,689 WE HAVE THE SAME EFFECT BUT WE 1370 00:55:32,689 --> 00:55:35,859 DID NOT GET THE DPLIEWTA MINE 1371 00:55:35,859 --> 00:55:36,426 BUT-- 1372 00:55:36,426 --> 00:55:36,727 RIGHT. 1373 00:55:36,727 --> 00:55:38,862 BUT IF WHEN YOU TOUCH THE TCA 1374 00:55:38,862 --> 00:55:39,863 TIKELE YOU'RE GETTING THE SAME 1375 00:55:39,863 --> 00:55:40,097 EFFECT. 1376 00:55:40,097 --> 00:55:42,165 IT MAY BE THAT IT'S METABOLISM 1377 00:55:42,165 --> 00:55:42,999 IN GENERAL THAT'S REQUIRED AND 1378 00:55:42,999 --> 00:55:45,969 IT DOESN'T HAVE TO BE DPLI 1379 00:55:45,969 --> 00:55:47,404 COLSIS PER SE, BUT IT CAN BE ALL 1380 00:55:47,404 --> 00:55:50,374 OF THESE THINGS FITTING IN,. 1381 00:55:50,374 --> 00:55:51,908 >> I MEAN I CAN GIVE YOU 1382 00:55:51,908 --> 00:55:52,976 PROTOCOLS FOR DON TREATMENT FOR 1383 00:55:52,976 --> 00:55:54,111 YOUR MICE IF YOU'RE INTERESTED. 1384 00:55:54,111 --> 00:55:55,579 >> THAT WOULD BE NICE, YES. 1385 00:55:55,579 --> 00:55:56,546 THAT COULD BE POSSIBLE. 1386 00:55:56,546 --> 00:55:58,248 THAT COULD BE POSSIBLE, THAT 1387 00:55:58,248 --> 00:56:01,118 THEY JUST NEED THE ENERGY, 1388 00:56:01,118 --> 00:56:03,553 ENOUGH ENERGY TO FEED THIS 1389 00:56:03,553 --> 00:56:04,588 PATHWAY AND DISCOAGULATION, 1390 00:56:04,588 --> 00:56:05,589 THAT'S A POSSIBILITY. 1391 00:56:05,589 --> 00:56:07,858 BUT WE DID NOT CHECK ON DPLIEWTA 1392 00:56:07,858 --> 00:56:08,392 MINE IN PARTICULAR. 1393 00:56:08,392 --> 00:56:09,092 >> IT WOULD BE INTERESTING ALSO 1394 00:56:09,092 --> 00:56:10,961 TO SEE IF YOU HAVE SWITCHES 1395 00:56:10,961 --> 00:56:12,796 BETWEEN SOME OF THE METABOLIC 1396 00:56:12,796 --> 00:56:14,464 PATHWAYS AND THEN JUST REALLY 1397 00:56:14,464 --> 00:56:16,633 QUICKLY BECAUSE YOU'RE ATP DATA 1398 00:56:16,633 --> 00:56:17,968 WAS REALLY INTERESTING, DO YOU 1399 00:56:17,968 --> 00:56:20,404 KNOW IF IF YOU HAVE AN 1400 00:56:20,404 --> 00:56:23,140 UPREGULATION OF ANY OF THE 1401 00:56:23,140 --> 00:56:24,541 NUCLEOTIDE TRANSPORTERS IF ARE 1402 00:56:24,541 --> 00:56:25,609 YOURSELF, LIKE ANTI1 OR 2, 1403 00:56:25,609 --> 00:56:27,911 BECAUSE IT HAS TO BE GETTING 1404 00:56:27,911 --> 00:56:29,112 INTO YOURSELF SOMEHOW SO HOW IS 1405 00:56:29,112 --> 00:56:29,846 THAT AFFECTING BECAUSE IT SEEMS 1406 00:56:29,846 --> 00:56:31,748 LIKE YOU HAVE A FEEDBACK 1407 00:56:31,748 --> 00:56:32,015 MECHANISM. 1408 00:56:32,015 --> 00:56:32,983 >> SO DISCIPLINARY NOT LOOK AT 1409 00:56:32,983 --> 00:56:34,317 THOSE ONCE BUT I WILL DEFINITELY 1410 00:56:34,317 --> 00:56:36,219 DO IT BECAUSE THAT COULD BE A 1411 00:56:36,219 --> 00:56:36,620 MECHANISM. 1412 00:56:36,620 --> 00:56:38,455 THAT COULD BE A MECHANISM. 1413 00:56:38,455 --> 00:56:41,057 SO WHAT I CAN SAY THAT FOR 1414 00:56:41,057 --> 00:56:45,595 EXAMPLE, THE LEVEL OF NTP 1, IS 1415 00:56:45,595 --> 00:56:46,730 ACTUALLY UPREGULATED IN KNOCKOUT 1416 00:56:46,730 --> 00:56:49,132 MICE SO IT'S SOMEHOW SPECIFIC ON 1417 00:56:49,132 --> 00:56:52,335 CD73, BUT WE DID NOT LOOK AT 1418 00:56:52,335 --> 00:56:53,136 OTHER TRANSPORTERS. 1419 00:56:53,136 --> 00:56:56,573 RIGHT, BECAUSE IT CAN BE A 1420 00:56:56,573 --> 00:57:00,043 CD973AXIS BUT IT CAN ALSO BE A 1421 00:57:00,043 --> 00:57:01,244 TRANSPORTER ITSELF. 1422 00:57:01,244 --> 00:57:01,578 CORRECT. 1423 00:57:01,578 --> 00:57:06,183 THANK YOU VERY MUCH. 1424 00:57:06,183 --> 00:57:06,483 GREAT. 1425 00:57:06,483 --> 00:57:16,793 [ APPLAUSE ]