1 00:00:11,440 --> 00:00:13,640 Welcome to the Clinical Center Grand Rounds, 2 00:00:13,640 --> 00:00:17,440 a weekly series of educational lectures for physicians and 3 00:00:17,440 --> 00:00:20,080 health care professionals broadcast from the Clinical 4 00:00:20,080 --> 00:00:23,040 Center at the National Institutes of Health in 5 00:00:23,040 --> 00:00:24,840 Bethesda, MD. 6 00:00:24,840 --> 00:00:28,400 The NIH Clinical Center is the world's largest hospital totally 7 00:00:28,400 --> 00:00:32,080 dedicated to investigational research and leads the global 8 00:00:32,080 --> 00:00:35,040 effort in training today's investigators and discovering 9 00:00:35,040 --> 00:00:37,200 tomorrow's cures. 10 00:00:37,200 --> 00:00:46,520 Learn more by visiting us online at http://clinicalcenter.nih.gov 11 00:00:46,520 --> 00:00:48,160 >>I'M VERY, VERY HONORED TO 12 00:00:48,160 --> 00:00:55,600 INTRODUCE OUR SPEAKER TODAY, 13 00:00:55,600 --> 00:00:57,400 DR. JANIS ABKOWITZ, LONG TIME 14 00:00:57,400 --> 00:00:59,520 HEAD OF HEMATOLOGY IN 15 00:00:59,520 --> 00:01:00,120 WASHINGTON, SEATTLE. 16 00:01:00,120 --> 00:01:01,640 SHE EARNED HER MEDICAL DEGREE 17 00:01:01,640 --> 00:01:03,640 FROM HARVARD MEDICAL SCHOOL AND 18 00:01:03,640 --> 00:01:05,680 COMPLETED RESIDENCY IN INTERNAL 19 00:01:05,680 --> 00:01:07,040 MEDICINE AND HEMATOLOGY AND 20 00:01:07,040 --> 00:01:08,120 ONCOLOGY TRAINING AT THE 21 00:01:08,120 --> 00:01:10,880 UNIVERSITY OF WASHINGTON AND 22 00:01:10,880 --> 00:01:11,840 FRED HUTCHINSON CANC 23 00:01:11,840 --> 00:01:13,720 E-PRESCRIBING RESEARCH CENTER. 24 00:01:13,720 --> 00:01:15,240 SHE'S REMAINED AT THE UNIVERSITY 25 00:01:15,240 --> 00:01:17,320 OF WASHINGTON AS A FACULTY 26 00:01:17,320 --> 00:01:17,800 MEMBER EVER SINCE. 27 00:01:17,800 --> 00:01:19,880 SHE IS A PHYSICIAN SCIENTIST WHO 28 00:01:19,880 --> 00:01:23,520 FOCUSES ON THE MOLECULAR AND 29 00:01:23,520 --> 00:01:26,400 CELLULAR DEFENSE IN RED CELL 30 00:01:26,400 --> 00:01:27,960 POTENTIATION AND SHE'S MADE 31 00:01:27,960 --> 00:01:29,720 MANY, MANY CONTRIBUTIONS 32 00:01:29,720 --> 00:01:31,480 INCLUDING QUANTITATIVE 33 00:01:31,480 --> 00:01:34,520 UNDERSTANDING OF HEMATOPOIETIC 34 00:01:34,520 --> 00:01:37,120 DYNAMICS, BEGINNING WITH 35 00:01:37,120 --> 00:01:37,960 X-LISTENED ISOILLUMINATIC 36 00:01:37,960 --> 00:01:39,640 EXPRESSION TO MODEL STEM CELL 37 00:01:39,640 --> 00:01:40,960 MODEL DYNAMICS. 38 00:01:40,960 --> 00:01:43,040 SHE ADENTIFIED A CLONE THE 39 00:01:43,040 --> 00:01:45,280 CELLULAR HEME EXPORTER AND GONE 40 00:01:45,280 --> 00:01:49,240 TO TO SEMINAL STUDYINGS IN HOW 41 00:01:49,240 --> 00:01:51,520 HEME AND PRODUCTION ARE RECORDED 42 00:01:51,520 --> 00:01:53,760 IN RED BLOOD CELL MATURATION, 43 00:01:53,760 --> 00:01:56,760 AND ANEMIA, IN A NUMBER OF HUMAN 44 00:01:56,760 --> 00:01:57,000 DISEASES. 45 00:01:57,000 --> 00:02:06,280 JAN HAS ALSO BEEN A FANTASTIC 46 00:02:06,280 --> 00:02:08,400 MENTOR--TO MANY OTHER WOMEN IN 47 00:02:08,400 --> 00:02:15,200 HEMATOLOGY WHO CHANGED THE FACE 48 00:02:15,200 --> 00:02:19,960 OF OUR INDUSTRY. 49 00:02:19,960 --> 00:02:21,160 DR. ABKOWITZ HAS SERVED AS 50 00:02:21,160 --> 00:02:22,800 MULTIPLE ROLES AT THE NIH 51 00:02:22,800 --> 00:02:24,640 INCLUDING BOARD OF SCIENTIFIC 52 00:02:24,640 --> 00:02:25,360 COUNSELORS AT NATIONAL HEART 53 00:02:25,360 --> 00:02:28,520 LUNG AND BLOOD INSTITUTE, THE 54 00:02:28,520 --> 00:02:32,200 TITLE OF DR. ABKOWITZ'S 55 00:02:32,200 --> 00:02:33,720 PRESENTATION IS ANEMIA FROM THE 56 00:02:33,720 --> 00:02:34,200 PERSPECTIVE OF IRON AND HEME. 57 00:02:34,200 --> 00:02:35,360 PLEASE JOIN ME IN WELCOMING 58 00:02:35,360 --> 00:02:36,000 DR. JAN ABKOWITZ. 59 00:02:36,000 --> 00:02:40,640 >> SO THANKS, SO MUCH, CINDY FOR 60 00:02:40,640 --> 00:02:41,280 THAT WONDERFUL INTRODUCTION. 61 00:02:41,280 --> 00:02:43,480 IT'S JUST A PLEASURE TO BE HERE 62 00:02:43,480 --> 00:02:47,040 >> SO WHAT I WANT TO TALK ABOUT 63 00:02:47,040 --> 00:02:49,160 TODAY IS ANEMIA FROM THE 64 00:02:49,160 --> 00:02:52,000 PERSPECTIVE OF IRON AND HEME, 65 00:02:52,000 --> 00:02:53,600 IT'S OFTEN TALKED ABOUT FROM I 66 00:02:53,600 --> 00:02:54,520 SCIENCE PERSPECTIVE OF GLOBEIN 67 00:02:54,520 --> 00:02:58,320 AND THIS IS A DIFFERENT 68 00:02:58,320 --> 00:02:58,800 PERSPECTIVE ON ANEMIA. 69 00:02:58,800 --> 00:03:00,080 I'M A CONSULTANT TO THIS 70 00:03:00,080 --> 00:03:01,480 MEDICINE WHICH IS A SMALL START 71 00:03:01,480 --> 00:03:03,720 UP BUT HAVE NO OTHER FINANCIAL 72 00:03:03,720 --> 00:03:05,920 DISCLOSURES, BUT I'M HAPPY TO 73 00:03:05,920 --> 00:03:09,000 DISCLOSE THAT I'M ABSOLUTELY 74 00:03:09,000 --> 00:03:09,960 PASSIONATE ABOUT BI-DIRECTIONAL 75 00:03:09,960 --> 00:03:10,240 TRANSLATION. 76 00:03:10,240 --> 00:03:12,880 AND SO OVER MY LIFE, I'VE BEEN 77 00:03:12,880 --> 00:03:16,000 WORKING ON CAT-MOUSE MODELS, 78 00:03:16,000 --> 00:03:17,080 CELLULAR MODELS OF HEMEAT O 79 00:03:17,080 --> 00:03:21,360 POETICESIS AND HOW THAT MIGHT 80 00:03:21,360 --> 00:03:25,040 APPLY TO CHILDREN WITH ANEMIA 81 00:03:25,040 --> 00:03:27,120 AND ADULTS WITH MILD DISPLACIA 82 00:03:27,120 --> 00:03:27,760 AND ADULTS IN BETWEEN. 83 00:03:27,760 --> 00:03:29,080 AND I THINK THAT OBSERVATIONS 84 00:03:29,080 --> 00:03:30,400 AND PATIENTS SHOULD DIRECT WHAT 85 00:03:30,400 --> 00:03:32,000 HAPPENS IN THE LABORATORY AND 86 00:03:32,000 --> 00:03:34,440 VICE VERSA AND HOPE TO CONVINCE 87 00:03:34,440 --> 00:03:39,200 YOU OF THAT TODAY. 88 00:03:39,200 --> 00:03:41,160 SO AS LEARNING OBJECTIVES, I 89 00:03:41,160 --> 00:03:43,600 WANT YOU TO UNDERSTAND HOW ORON 90 00:03:43,600 --> 00:03:44,800 AND HEME REGULATE RED CELL 91 00:03:44,800 --> 00:03:46,840 PRODUCTION, WHAT ARE THE 92 00:03:46,840 --> 00:03:52,200 CONSEQUENCES OF ANEMIA, AND WHY 93 00:03:52,200 --> 00:03:54,760 INEFFECTIVE ERYTH ROUGH ATOM 94 00:03:54,760 --> 00:03:56,040 POETICESIS AND WHICH RED CELLS 95 00:03:56,040 --> 00:03:59,160 DIE AND DON'T MAKE IT OUT TO BE 96 00:03:59,160 --> 00:04:06,400 RED CELLS IS WHAT 97 00:04:06,400 --> 00:04:08,400 ERYTHROPOIESIS, AND THAL 98 00:04:08,400 --> 00:04:08,680 PHENOTYPE. 99 00:04:08,680 --> 00:04:11,160 SO FIRST ABOUT HEME, IRON, 100 00:04:11,160 --> 00:04:12,960 ANEMIA AND RED CELLS. 101 00:04:12,960 --> 00:04:17,040 THE GLOBAL PRESENCE OF ANEMIA IS 102 00:04:17,040 --> 00:04:18,640 QUITE HIGH, 33%. 103 00:04:18,640 --> 00:04:22,520 ACCOUNTING FOR OVER 68 MILLION 104 00:04:22,520 --> 00:04:23,680 YEARS OF DISABILITY. 105 00:04:23,680 --> 00:04:25,600 THIS INCLUDES SOME VERY CLASSIC 106 00:04:25,600 --> 00:04:30,320 STUDIES SUCH AS REFERENCED BELOW 107 00:04:30,320 --> 00:04:33,720 WHICH IS IN SRI LANKA, AND THEY 108 00:04:33,720 --> 00:04:41,920 HAVE A HIGH PREVALENCE OF WORM 109 00:04:41,920 --> 00:04:44,800 AND LOSE PRODUCTION IN THE GI 110 00:04:44,800 --> 00:04:46,560 TRACT AND THERE'S MANY OTHER 111 00:04:46,560 --> 00:04:47,880 EXAMPLES LIKE THAT IN THE 112 00:04:47,880 --> 00:04:49,200 DEVELOPING WORLD AND THEN FOR 113 00:04:49,200 --> 00:04:52,560 US, THERE'S EXAMPLES SUCH AS MDS 114 00:04:52,560 --> 00:04:54,200 OR MILD DISPLASTIC SYNDROME 115 00:04:54,200 --> 00:04:56,280 WHICH IS ANY O PLASTIC OR CLONAL 116 00:04:56,280 --> 00:05:00,400 PROCESS THAT STARTS IN THE 117 00:05:00,400 --> 00:05:02,400 HEMATOPOIETIC STEM OR 118 00:05:02,400 --> 00:05:04,000 MULTIPOTENT PROGENITOR CELL THAT 119 00:05:04,000 --> 00:05:05,360 RESULTS IN CYTOPENIA. 120 00:05:05,360 --> 00:05:07,200 THE MEDIAN AGE OF DIAGNOSIS IS 121 00:05:07,200 --> 00:05:12,600 71 YEARS AND APPROXIMATELY 81% 122 00:05:12,600 --> 00:05:15,000 OF PATIENTS WITH OR 80% OF 123 00:05:15,000 --> 00:05:16,920 MAISHTS WITH MDS ARE DIAGNOSED 124 00:05:16,920 --> 00:05:18,200 BECAUSE OF ANEMIA. 125 00:05:18,200 --> 00:05:20,320 THAT ANEMIA IS GENERALLY 126 00:05:20,320 --> 00:05:21,400 MACROSIDIC WHICH MEANS THE RED 127 00:05:21,400 --> 00:05:27,600 CELLS ARE BIG AND IT'S USUALLY 128 00:05:27,600 --> 00:05:29,120 DUE TO ERYTHROPOIESIS, AND NOW 129 00:05:29,120 --> 00:05:30,680 ANEMIA IS A MAJOR CAUSE OF 130 00:05:30,680 --> 00:05:35,120 MORBIDITY IN THIS SETTING AS 131 00:05:35,120 --> 00:05:35,320 WELL. 132 00:05:35,320 --> 00:05:37,400 FOR EXAMPLE, THERE ARE MANY 133 00:05:37,400 --> 00:05:38,200 CLINICAL ASSOCIATIONS OF ANEMIA 134 00:05:38,200 --> 00:05:39,880 IN OLDER PERSONS AND THE MDS, 135 00:05:39,880 --> 00:05:42,240 DISEASE OF OLDER PERSONS, 136 00:05:42,240 --> 00:05:43,880 INCREASED FALLS, INCREASED RATES 137 00:05:43,880 --> 00:05:45,120 OF MAJOR DEPRESSION, INCREASED 138 00:05:45,120 --> 00:05:46,800 RATE OF HOSPITALIZATION AND 139 00:05:46,800 --> 00:05:49,480 LONGER DURATION OF 140 00:05:49,480 --> 00:05:50,960 HOSPITALIZATION, DECREASED 141 00:05:50,960 --> 00:05:52,520 MOBILITY AND BONE DENSITY AND 142 00:05:52,520 --> 00:05:55,320 SKELETAL MASK AND DECREASED 143 00:05:55,320 --> 00:05:56,720 FUNCTION, COGNITION AND 144 00:05:56,720 --> 00:05:58,360 MORTALITY SO ANEMIA IS BROADLY 145 00:05:58,360 --> 00:05:59,400 SOMETHING INTRIGUING TO THINK 146 00:05:59,400 --> 00:06:01,000 ABOUT FROM A MORE BASIC 147 00:06:01,000 --> 00:06:02,600 PERSPECTIVE OF HOW ARE RED CELLS 148 00:06:02,600 --> 00:06:07,080 MADE OR WHAT ERYTHROPOIESIS AND 149 00:06:07,080 --> 00:06:09,440 HOW IS IT PUT TOGETHER? 150 00:06:09,440 --> 00:06:11,040 AND VERY INTRIGUING TO ME QUITE 151 00:06:11,040 --> 00:06:14,400 EARLY ON IN MY STUDIES IS THAT 152 00:06:14,400 --> 00:06:15,800 ERYTHROPOIESIS HAS RAPID 153 00:06:15,800 --> 00:06:17,280 KINETICS. 154 00:06:17,280 --> 00:06:18,400 SO PEOPLE MAKE 2.3 MILLION RED 155 00:06:18,400 --> 00:06:20,440 CELLS A SECOND AND THIS CAN 156 00:06:20,440 --> 00:06:22,680 INCREASE 5-10 FOLD IN RESPONSE 157 00:06:22,680 --> 00:06:26,800 TO ANEMIA WHICH IS WOULD BE 158 00:06:26,800 --> 00:06:31,200 MODERATED BY THE HEME COUNT. 159 00:06:31,200 --> 00:06:34,760 AND SO, THERE ARE 270 MILLION 160 00:06:34,760 --> 00:06:35,920 HEMOGLOBIN MOLECULES PER CELL 161 00:06:35,920 --> 00:06:42,000 AND EACH HAVE 4 HEME AND 4 162 00:06:42,000 --> 00:06:43,560 GLOBIN CHAINS AND TOGETHER THAT 163 00:06:43,560 --> 00:06:54,000 MAKES HEME OR HEME MOETYS, BEING 164 00:06:54,000 --> 00:06:56,320 2.5 TIMES 10 TO THE TENTH FOLD 165 00:06:56,320 --> 00:06:58,240 SO LARGE QUANTITIES ARE NEEDED 166 00:06:58,240 --> 00:07:00,000 AND NEEDED QUICKLY AS RED BLOOD 167 00:07:00,000 --> 00:07:02,480 CELLS MATURE BUT THERE'S A 168 00:07:02,480 --> 00:07:05,720 PROBLEM BECAUSE FREE HEME IS 169 00:07:05,720 --> 00:07:09,280 TOXIC, AT LEAST IF ROS, IT LEADS 170 00:07:09,280 --> 00:07:10,400 TO OXIDATION, AND CELL DEATH, SO 171 00:07:10,400 --> 00:07:12,120 THEY HAVE TO CONTROL THIS AND 172 00:07:12,120 --> 00:07:14,560 HEME IT OR PROCESS IT OR EXPORT 173 00:07:14,560 --> 00:07:18,640 IT IN ORDER TO BE SAFE. 174 00:07:18,640 --> 00:07:24,000 SO HERE'S THE BUFFER CELLS, HEME 175 00:07:24,000 --> 00:07:28,600 IS A CHEMICAL CHELATE, AND 176 00:07:28,600 --> 00:07:31,200 GLOBEIN IS A PROTEIN MADE IN THE 177 00:07:31,200 --> 00:07:35,000 SAME QUANTITY AND TIME AND IT'S 178 00:07:35,000 --> 00:07:36,280 PRODUCED BY TRANSCRIPTION AND 179 00:07:36,280 --> 00:07:37,440 TRANSLATION, SO PARTICULARLY 180 00:07:37,440 --> 00:07:40,000 UNDER THIS DEMAND OF THE 181 00:07:40,000 --> 00:07:44,360 KINETICS CANNOT RELY ON 182 00:07:44,360 --> 00:07:48,000 TRANSCRIPTION AND TRANSLATION, 183 00:07:48,000 --> 00:07:49,200 CELLS NEEDED ALTERNATE SOLUTIONS 184 00:07:49,200 --> 00:07:50,320 AND THAT HAS DRIVEN THE 185 00:07:50,320 --> 00:07:53,120 QUESTIONS I HAVE ASKED IN THE 186 00:07:53,120 --> 00:07:53,560 LABORATORY SETTING. 187 00:07:53,560 --> 00:07:56,000 SO WHAT I PLAN TO DO IS BREATHE 188 00:07:56,000 --> 00:07:59,080 IN STUDIES OF THE SYNDROME, 189 00:07:59,080 --> 00:08:00,200 PARTICULAR SUBTYPE OF SYNDROME 190 00:08:00,200 --> 00:08:02,840 THAT RESULTS FROM ISOLATED 191 00:08:02,840 --> 00:08:06,160 DELETION OF CHROMOSOME 5 OR DEL5 192 00:08:06,160 --> 00:08:09,400 QMDS AND A GENERAL MACROSITTIC 193 00:08:09,400 --> 00:08:11,160 ANEMIA, DBA, I WILL SHOW THAT 194 00:08:11,160 --> 00:08:14,440 THESE ARE INEFFECTIVE AND 195 00:08:14,440 --> 00:08:15,320 MACROSITTIC ANEMIAS, MEANING THE 196 00:08:15,320 --> 00:08:19,320 SEDENTARY CELLS ARE BIG AND THEY 197 00:08:19,320 --> 00:08:20,720 RESULT FROM EXCESS HEAT AND I 198 00:08:20,720 --> 00:08:22,720 WILL ARGUE WHY THIS IS 199 00:08:22,720 --> 00:08:25,240 GENERALIZABLE TO MOST MDS ANEMIA 200 00:08:25,240 --> 00:08:28,760 AND PROVIDE INSIGHTS TO HEMEAT O 201 00:08:28,760 --> 00:08:30,000 POETIS, ERYTHROPOIESIS AND NEW 202 00:08:30,000 --> 00:08:32,000 TARGETS FOR THERAPY AND LASTLY 203 00:08:32,000 --> 00:08:34,200 SHOW HOW THESE CAN BE COMBINED 204 00:08:34,200 --> 00:08:35,480 TO ANSWER PHYSIOLOGIC QUESTIONS 205 00:08:35,480 --> 00:08:37,480 THAT HAVE A CLINICAL IMPACT. 206 00:08:37,480 --> 00:08:38,880 SO I'M GOING TO ACTUALLY START 207 00:08:38,880 --> 00:08:41,000 WITH THE ANSWER AND THEN PRESIDE 208 00:08:41,000 --> 00:08:43,120 DATA AND EXTENSION OF THAT AND 209 00:08:43,120 --> 00:08:46,160 SO WHEN 1 THINKS ABOUT RED CELL 210 00:08:46,160 --> 00:08:48,840 SIEVE RENTIATION, IT STARTS IN 211 00:08:48,840 --> 00:08:51,040 THE VERY EARLY STAGE OF A 212 00:08:51,040 --> 00:08:53,360 COMMITTED CELL CALLED A BFUE 213 00:08:53,360 --> 00:08:56,600 SHOWN HERE AND UNDER THE 214 00:08:56,600 --> 00:09:02,720 INFLUENCE OF GADA 1, THE 215 00:09:02,720 --> 00:09:03,480 ERYTHROPOIESIS IS UPREGULATED 216 00:09:03,480 --> 00:09:05,640 AND THAT ALLOWS THEM TO 217 00:09:05,640 --> 00:09:06,960 TRANSDUCE RECEPTIONOR 1 WHICH 218 00:09:06,960 --> 00:09:08,720 ALLOWS IRON INTO CELLS. 219 00:09:08,720 --> 00:09:14,960 THE IRON BINDS TO IRON PROTEINS, 220 00:09:14,960 --> 00:09:18,800 INDUCES A LAS 2, AND THAT IS THE 221 00:09:18,800 --> 00:09:21,080 FIRST REGULATING STEM IN HEMEAT 222 00:09:21,080 --> 00:09:24,000 O SYNTHS SIS, AND HEME SENTH SIS 223 00:09:24,000 --> 00:09:25,280 INTENSIFYS AND THEN HEME IS 224 00:09:25,280 --> 00:09:27,360 ACTUALLY THE INDUCER OF GLOBEIN 225 00:09:27,360 --> 00:09:28,200 TRANSCRIPTION AND TRANSLATION 226 00:09:28,200 --> 00:09:32,920 AND IT DOES BOTH BY REPRESSING 227 00:09:32,920 --> 00:09:36,120 REPRESSORS, SO AS SOON AS HE'S 228 00:09:36,120 --> 00:09:36,960 PRESENT GLOBEIN TRANSCRIPTION 229 00:09:36,960 --> 00:09:37,680 AND TRANSLATION CAN OCCUR BUT 230 00:09:37,680 --> 00:09:39,800 THIS IS A KINETIC PROCESS AND IT 231 00:09:39,800 --> 00:09:41,800 MEANS THERE'S A TIME DURING 232 00:09:41,800 --> 00:09:44,800 DEFERENTIATION RIGHT AT THE CFUE 233 00:09:44,800 --> 00:09:47,720 PROARITHROUGH ATOM BLAST STAGE 234 00:09:47,720 --> 00:09:48,760 WHERE HEME EXCEEDS GLOBEIN AND 235 00:09:48,760 --> 00:09:50,520 THE CELL NEEDS TO DO SOMETHING 236 00:09:50,520 --> 00:09:52,160 ABOUT THAT BECAUSE HEME'S TOXIC 237 00:09:52,160 --> 00:09:53,680 AND IT HAS TO BE CONTROLLED AND 238 00:09:53,680 --> 00:09:56,960 WHAT IT DOES IS IT EXPORTS IT 239 00:09:56,960 --> 00:10:00,920 THROUGH FLBCR AND THIS PROTECTS 240 00:10:00,920 --> 00:10:07,160 CFUE BLASTINGS FROM HEME 241 00:10:07,160 --> 00:10:07,440 TOXICITIES. 242 00:10:07,440 --> 00:10:09,200 SO WHEY WANT TO POINT OUT WITH 243 00:10:09,200 --> 00:10:11,360 THIS IS THAT SOMEONE CAN TRACK 244 00:10:11,360 --> 00:10:12,400 THE DIFFERENTIATION WHICH ALLOWS 245 00:10:12,400 --> 00:10:17,400 IRON IN AND LATE MARKER FOR AN A 246 00:10:17,400 --> 00:10:19,400 IN HUMAN AND FOR 119 IN MOUSE 247 00:10:19,400 --> 00:10:21,680 AND YOU CAN HAVE WHERE IT IS 248 00:10:21,680 --> 00:10:22,880 OFF, RECEPTOR IS ON AND THEY'RE 249 00:10:22,880 --> 00:10:25,640 IN THIS STAGE OF DIFFERENTIATION 250 00:10:25,640 --> 00:10:27,160 WHERE THEY'RE BOTH EXPRESSED AND 251 00:10:27,160 --> 00:10:34,600 THEY FEEL LIKE A FOREIGN EXPRESS 252 00:10:34,600 --> 00:10:35,560 AND THEY CAN LAUNCH ACROSS THE 253 00:10:35,560 --> 00:10:36,960 SEQUENCE AND I WILL SHOW YOU 254 00:10:36,960 --> 00:10:39,000 THIS HERE, WHICH OVERLAPS CD671 255 00:10:39,000 --> 00:10:41,200 AND WE USE IN A NUMBER OF 256 00:10:41,200 --> 00:10:43,800 STUDIES IN INVITRO, BECAUSE CD71 257 00:10:43,800 --> 00:10:48,400 BEING A RECEPTOR FOR TRANSFERIN 258 00:10:48,400 --> 00:10:50,960 IS MODULATED BY IRON AND IRON 259 00:10:50,960 --> 00:10:52,960 INVITTO CAN VARY DEPENDING ON 260 00:10:52,960 --> 00:10:54,560 CULTURE CONDITIONS AND CD36 DOES 261 00:10:54,560 --> 00:10:56,560 NOT SEE IT BECOMES A FAIR 262 00:10:56,560 --> 00:10:59,800 MEASURE OF DIFFERENTIATION 263 00:10:59,800 --> 00:11:00,240 ACROSS THE SEQUENCE. 264 00:11:00,240 --> 00:11:02,560 SO JUST A BIT OF BACKGROUNDOT 265 00:11:02,560 --> 00:11:04,880 PHYSIOLOGY OF DBA AND MY O 266 00:11:04,880 --> 00:11:05,760 DISPLASTIC SYNDROME. 267 00:11:05,760 --> 00:11:09,320 SO DBA IS A CONGENITAL AND 268 00:11:09,320 --> 00:11:11,320 MACROSITTIC ANEMIA AND 25% OF 269 00:11:11,320 --> 00:11:14,640 DBA CASES RESULT IN HAPPEN LUNG 270 00:11:14,640 --> 00:11:17,440 CANCER ENSEL ACTIVITY AND 65% 271 00:11:17,440 --> 00:11:18,680 FROM HAPPEN LO INSUFFICIENCIES 272 00:11:18,680 --> 00:11:20,880 IN 19 OTHER RIBOSOMAL PROTEINS, 273 00:11:20,880 --> 00:11:23,400 SO HAVE YOU EXACTLY THE SAME 274 00:11:23,400 --> 00:11:25,960 CLINICAL PHENOTYPE WITH 20 275 00:11:25,960 --> 00:11:31,120 DIFFERENT RIBOSOMAL PROTEINS, 276 00:11:31,120 --> 00:11:35,720 BEING HAPLO-INSUFFICIENCY AND 277 00:11:35,720 --> 00:11:38,360 INTERESTINGLY THE MACRO SITTIC 278 00:11:38,360 --> 00:11:41,360 ANEMIA OF MDS IS TESTED FOR THE 279 00:11:41,360 --> 00:11:44,720 ACQUIRES MUTATION AND 280 00:11:44,720 --> 00:11:45,240 HAPLOINSUFFICIENCY RPS14. 281 00:11:45,240 --> 00:11:49,440 SO WHEN THESE ARE IN THE 282 00:11:49,440 --> 00:11:51,000 MDS SHARE A CLINICAL PHENOTYPE, 283 00:11:51,000 --> 00:11:52,040 SLOWED OR IMPAIRED TRANSLATION 284 00:11:52,040 --> 00:11:59,160 COULD BE THROUGH TO THEIR 285 00:11:59,160 --> 00:11:59,680 PATHOGENESIS. 286 00:11:59,680 --> 00:12:05,640 I WILL SHOW YOU DATA AGAIN ABOUT 287 00:12:05,640 --> 00:12:06,600 SOPHISTICATED AGAIN, BFUE, AND 288 00:12:06,600 --> 00:12:07,560 NONTO RED CELLS, CAN YOU MARK 289 00:12:07,560 --> 00:12:10,360 THEM WITH THE DIFFERENT MARKERS, 290 00:12:10,360 --> 00:12:13,120 HEME SYNTH TIS INTENSIFYS AT THE 291 00:12:13,120 --> 00:12:14,680 CFUE STAGE AND GLOBE'S SYNTH TIS 292 00:12:14,680 --> 00:12:16,120 COMES IN LATER AND THEREYA A 293 00:12:16,120 --> 00:12:18,640 PERIOD OF TIME WHEN HEME ECSEATS 294 00:12:18,640 --> 00:12:20,160 GLOBEIN AND THEY COMPENSATE FOR 295 00:12:20,160 --> 00:12:22,680 THE RED CELLS FOR THEM TO BE 296 00:12:22,680 --> 00:12:23,040 PERFECTLY FINE. 297 00:12:23,040 --> 00:12:25,840 BUT WHAT IF YOU HAD POOR ROBO 298 00:12:25,840 --> 00:12:27,360 STUDIES OF MULTIPLE ENDOCRINAL 299 00:12:27,360 --> 00:12:28,480 ASSEMBLY AND POOR PROTEIN 300 00:12:28,480 --> 00:12:30,400 TRANSLATION, AND THEN THE 301 00:12:30,400 --> 00:12:32,200 PROTEIN BEING POORLY TRANSLATED 302 00:12:32,200 --> 00:12:34,440 BUT HEME AND A KEY LINK SO VERY 303 00:12:34,440 --> 00:12:36,800 LITTLE PROTEIN IS NEEDED FOR THE 304 00:12:36,800 --> 00:12:38,120 ENZYMES MADE OUT OF HEME SO YOU 305 00:12:38,120 --> 00:12:40,040 END UP WITH THE CIRCUMSTANCE 306 00:12:40,040 --> 00:12:42,000 WHERE THERE'S PLENTY OF HEME, 307 00:12:42,000 --> 00:12:43,600 ADEQUATE HEME AT AN APPROPRIATE 308 00:12:43,600 --> 00:12:45,800 TIME BUT WERE GLOBEIN IN HAVING 309 00:12:45,800 --> 00:12:47,400 THE AMOUNT OF GLOBEIN YOU NEED 310 00:12:47,400 --> 00:12:50,800 TO BIND THAT AMOUNT OF HEME IS 311 00:12:50,800 --> 00:12:52,400 DELAYED OR IMPAIRED. 312 00:12:52,400 --> 00:12:54,400 AND SO HEME EXCESS EXTENDS FOR A 313 00:12:54,400 --> 00:12:58,160 LONGER PERIOD OF TIME, HEME 314 00:12:58,160 --> 00:13:00,800 EXPORT VIA FLBCR IS NOT 315 00:13:00,800 --> 00:13:02,400 EFFICIENT AND CELLS BY AT THIS 316 00:13:02,400 --> 00:13:04,960 STAGE AT THE STAGE OF 317 00:13:04,960 --> 00:13:05,320 DIFFERENTIATION. 318 00:13:05,320 --> 00:13:06,040 NOW SOME DATA FOR THAT. 319 00:13:06,040 --> 00:13:14,560 THIS IS FROM A PATIENT WITH DBA, 320 00:13:14,560 --> 00:13:15,480 HAPLOINSUFFICIENCY OF 19, THE 321 00:13:15,480 --> 00:13:17,800 MOST COMMON FORM AND SHE IS A 23 322 00:13:17,800 --> 00:13:20,320 YEAR-OLD WHO IS TRANSFUSION 323 00:13:20,320 --> 00:13:25,000 DEPENDENT, SO IS ON NO 324 00:13:25,000 --> 00:13:25,400 MEDICATION. 325 00:13:25,400 --> 00:13:26,400 HER MARROW HAS ADEQUATE NUMBERS 326 00:13:26,400 --> 00:13:28,600 OF STEM CELLS AND EARLY 327 00:13:28,600 --> 00:13:30,200 PROGENITOR CELLS AS SHOWN HERE 328 00:13:30,200 --> 00:13:30,920 COMPARED TO NORMAL, HOWEVER WHEN 329 00:13:30,920 --> 00:13:33,000 YOU LOOK AT THE EXPRESSION OF 330 00:13:33,000 --> 00:13:34,440 CD71 ON THE X-AXIS SO THE 331 00:13:34,440 --> 00:13:37,120 EXPRESSION OF TRANSFER AND 332 00:13:37,120 --> 00:13:38,880 RECEPTOR, ASOON AS CELLS EXPRESS 333 00:13:38,880 --> 00:13:40,560 A TRANSFER AND RECEPTOR THEY 334 00:13:40,560 --> 00:13:41,040 DIE. 335 00:13:41,040 --> 00:13:42,760 WHEN NORMAL CELLS EXPRESS A 336 00:13:42,760 --> 00:13:44,800 TRANSGERM RECEPTOR AND THEY 337 00:13:44,800 --> 00:13:48,800 MATURE, AS MATURE RED CELLS, AND 338 00:13:48,800 --> 00:13:50,520 SO, INVIVO, DEATH OCCURS AT JUST 339 00:13:50,520 --> 00:13:53,200 THE STAGE THAT WE WOULD PREDICT 340 00:13:53,200 --> 00:13:54,640 FROM MY LAST SLIDE. 341 00:13:54,640 --> 00:13:56,560 AND SO WHAT WE THOUGHT TO DO WAS 342 00:13:56,560 --> 00:13:59,800 A CULTURE MARROW CELLS FROM DBA 343 00:13:59,800 --> 00:14:01,840 PATIENTS FROM DELATWAL 5 MDS 344 00:14:01,840 --> 00:14:02,720 PATIENTS FROM NORMAL INDIVIDUALS 345 00:14:02,720 --> 00:14:04,680 AND CONTROLS AND FOLLOW THEIR 346 00:14:04,680 --> 00:14:06,520 ERYTHROPOIESIS INVITRO AND WHAT 347 00:14:06,520 --> 00:14:08,680 HAPPENS IN INROW AND YOU CAN 348 00:14:08,680 --> 00:14:11,600 KIND OF STABILIZE THESE CELLS SO 349 00:14:11,600 --> 00:14:12,840 THOUGH THEY'RE UNHEALTHY, THEY 350 00:14:12,840 --> 00:14:14,400 MATURE A BIT FURTHER AND YOU CAN 351 00:14:14,400 --> 00:14:15,600 QUERY THEM ABOUT LAWN MOWER'S 352 00:14:15,600 --> 00:14:17,080 GOING ON WITH A LOT MORE 353 00:14:17,080 --> 00:14:18,200 PRECISION AND SO HERE'S AN 354 00:14:18,200 --> 00:14:20,840 EXAMPLE OF THAT, THIS IS AGAIN A 355 00:14:20,840 --> 00:14:23,400 DBA PATIENT, DIFFERENT 1 THIS 1, 356 00:14:23,400 --> 00:14:26,560 CD36 GOING IN THIS DIRECTION, 357 00:14:26,560 --> 00:14:28,560 GLYCO 4IN A GOING IN THIS 358 00:14:28,560 --> 00:14:30,120 DIRECTION AND NORMAL MATURE FROM 359 00:14:30,120 --> 00:14:32,920 1, 2, 3, 4, LIKE THIS. 360 00:14:32,920 --> 00:14:35,360 THE DBA PATIENT APPEARS TO HAVE 361 00:14:35,360 --> 00:14:37,000 A BLOCK SOMEWHERE AROUND HERE IN 362 00:14:37,000 --> 00:14:38,080 THE MATURATION SEQUENCE AND NOT 363 00:14:38,080 --> 00:14:40,000 AS MANY CELLS THAT COME DOWN TO 364 00:14:40,000 --> 00:14:41,120 4, BUT IMPORTANTLY IF YOU LOOK 365 00:14:41,120 --> 00:14:42,960 BY STAGE IN THE NUMBER OF CELLS 366 00:14:42,960 --> 00:14:44,960 WHICH IS SHOWN BELOW, THAT STAGE 367 00:14:44,960 --> 00:14:46,840 1 FOR THE EARLIER CELLS HAS MORE 368 00:14:46,840 --> 00:14:48,720 OR LESS THE SAME NUMBER OF CELLS 369 00:14:48,720 --> 00:14:50,520 BUT THERE'S A REAL BLOCK HERE, 370 00:14:50,520 --> 00:14:52,000 THERE'S A CUT OFF, WHERE IT 371 00:14:52,000 --> 00:14:53,880 SEEMS TO BE DEATH BETWEEN STAGE 372 00:14:53,880 --> 00:14:57,280 1 AND STAGE 2 WHICH WOULD BE 373 00:14:57,280 --> 00:14:58,200 AGAIN THE CFUE PROERYTHROPOIESIS 374 00:14:58,200 --> 00:15:00,640 LEVEL AND THOSE CELLS THAT 375 00:15:00,640 --> 00:15:01,800 MAKE-THAT ACTUALLY MAKE IT TO 376 00:15:01,800 --> 00:15:05,080 STAGE 2, THEY ACTUALLY EXPAND 377 00:15:05,080 --> 00:15:05,280 OKAY. 378 00:15:05,280 --> 00:15:06,880 AND SO THE TAKE HOME FROM THIS 379 00:15:06,880 --> 00:15:09,640 IS THAT THERE SEEMS TO BE A HIT 380 00:15:09,640 --> 00:15:10,360 THAT'S VERY EARLY IN 381 00:15:10,360 --> 00:15:12,080 ERYTHROPOIESIS BUT IF YOU MAKE 382 00:15:12,080 --> 00:15:14,400 IT PAST THAT HIT THAT EXPANSION 383 00:15:14,400 --> 00:15:16,120 OF RED CELLS IS OKAY. 384 00:15:16,120 --> 00:15:17,680 AND SO THEN WE ASKED THE 385 00:15:17,680 --> 00:15:19,200 QUESTION, WHAT'S HAPPENING AT 386 00:15:19,200 --> 00:15:21,480 THE HIT, WHAT'S HAPPENING AT 387 00:15:21,480 --> 00:15:23,400 STAGE 1, AND WHAT IS THE 388 00:15:23,400 --> 00:15:24,880 CHARACTERISTICS OF THE CELLS AS 389 00:15:24,880 --> 00:15:26,640 STAGE 3 AND 4, THOSE HAVE MADE 390 00:15:26,640 --> 00:15:26,880 IT. 391 00:15:26,880 --> 00:15:30,520 HOW ARE THEY DIFFERENT FROM 392 00:15:30,520 --> 00:15:32,160 THOSE THAT DON'T? 393 00:15:32,160 --> 00:15:35,360 SO AT STAGE 1, THERE IS MORE 394 00:15:35,360 --> 00:15:40,800 HEME CONTENT IN DBA CELLS AND 395 00:15:40,800 --> 00:15:42,160 THOSE THAT MAKE IT THAT MAKE IT 396 00:15:42,160 --> 00:15:46,400 TO STAGE 3 OR 4, HAVE LESS ALAS2 397 00:15:46,400 --> 00:15:49,040 EXPRESSION AND MAKE LESS HEME OR 398 00:15:49,040 --> 00:15:50,800 HAVE MORE FLVCR EXPRESSION, 399 00:15:50,800 --> 00:15:51,200 EXPORT OR HEME. 400 00:15:51,200 --> 00:15:52,800 SO THOSE CELLS THAT ARE ABLE TO 401 00:15:52,800 --> 00:15:55,880 MAKE IT THROUGH ARE ABLE TO 402 00:15:55,880 --> 00:15:59,600 CONTROL HEME IN A WAY THAT 403 00:15:59,600 --> 00:16:02,600 ALLOWS THAT TO HAPPEN. 404 00:16:02,600 --> 00:16:06,600 NOW YOU COULD DEL5 Q MDS, SAME 405 00:16:06,600 --> 00:16:09,160 THING, THOSE THAT ARE AT STAGE 1 406 00:16:09,160 --> 00:16:10,120 HAVE EXTRA HEME CONTENT. 407 00:16:10,120 --> 00:16:12,800 THOSE THAT MAKE IT THROUGH THE 408 00:16:12,800 --> 00:16:15,800 BLOCK ARE ABLE TO CONTROL THEIR 409 00:16:15,800 --> 00:16:17,400 HEME SYNTHESIS OR IN THIS 410 00:16:17,400 --> 00:16:18,560 SITUATION HAVE SKY HIGH EXPORT 411 00:16:18,560 --> 00:16:24,000 OF HEME TO CONTROL THEIR HEME 412 00:16:24,000 --> 00:16:24,240 AMOUNT. 413 00:16:24,240 --> 00:16:26,000 WE THEN LOOK AT THIS A DIFFERENT 414 00:16:26,000 --> 00:16:26,200 WAY. 415 00:16:26,200 --> 00:16:28,360 WE LOOK AT CELLS OVER TIME AND 416 00:16:28,360 --> 00:16:30,000 CULTURE AND WHEN IT STARTS AT 417 00:16:30,000 --> 00:16:32,280 DAY 0 AND LOOKS UP TO DAY 18 AS 418 00:16:32,280 --> 00:16:33,680 RED CELLS MATURE, AND THIS IS 419 00:16:33,680 --> 00:16:37,800 LOOKING NOW AT PROTEIN FOR ALIS 420 00:16:37,800 --> 00:16:40,720 2 AND PROTEIN FOR GLOBEIN AND 421 00:16:40,720 --> 00:16:42,280 WHAT'S IMPORTANT TO POINT OUT IS 422 00:16:42,280 --> 00:16:45,880 THAT ONLY RED CELLS MAKE ALAS2, 423 00:16:45,880 --> 00:16:48,280 AS A SPECIFIC ENZYME AND ONLY 424 00:16:48,280 --> 00:16:50,640 RED CELLS MAKE GLOBEIN, IF THERE 425 00:16:50,640 --> 00:16:52,000 WAS A MACROPHAGE CONTAMINATION, 426 00:16:52,000 --> 00:16:54,240 A WHITE CELL THAT LEFT IN YOUR 427 00:16:54,240 --> 00:16:55,880 CELL CULTURE, IT WOULDN'T 428 00:16:55,880 --> 00:16:56,920 CONTRIBUTE TO GLOBEIN, SO 429 00:16:56,920 --> 00:16:57,800 THEREFORE IF YOU TRACK 430 00:16:57,800 --> 00:17:08,680 INDIVIDUAL CULTURES IN TERMS OF 431 00:17:08,680 --> 00:17:09,760 WHETHER--IF YOU'RE TRACK BEING 432 00:17:09,760 --> 00:17:12,320 THOSE CELLS EVEN IF THAT CULTURE 433 00:17:12,320 --> 00:17:13,800 IS NOT ENTIRELY HOMOGEANIOUS. 434 00:17:13,800 --> 00:17:17,000 SO SHOWN HERE IN THIS EXAMPLE OF 435 00:17:17,000 --> 00:17:18,800 DAY 3, THERE'S ACTUALLY PLENTY 436 00:17:18,800 --> 00:17:20,040 OF HEME BEING MADE. 437 00:17:20,040 --> 00:17:21,800 IN THIS PARTICULAR EXAMPLE, 438 00:17:21,800 --> 00:17:23,520 THERE'S PROTEIN, SO MORE HEME 439 00:17:23,520 --> 00:17:24,960 SYNTHESIS IN THE DBA IN BLUE 440 00:17:24,960 --> 00:17:29,240 THAN THERE IS IN THE CONTROL 441 00:17:29,240 --> 00:17:31,120 THAT WAS 1 CONCUR EPTLY, 442 00:17:31,120 --> 00:17:33,800 HOWEVER, AS SHOWN BELOW, THERE'S 443 00:17:33,800 --> 00:17:37,000 A DELAY IN THE HEMOGLOBIN 444 00:17:37,000 --> 00:17:37,800 PRODUCTION. 445 00:17:37,800 --> 00:17:40,080 WOW, SO THAT THE DISCREPANCY 446 00:17:40,080 --> 00:17:41,520 HERE BETWEEN HEMOGLOBIN IS QUITE 447 00:17:41,520 --> 00:17:44,600 PRONOUNCED AND IF I'M LOOKING AT 448 00:17:44,600 --> 00:17:48,600 THE ASSAY OF THAT CULTURE ON DAY 449 00:17:48,600 --> 00:17:51,200 7 WHEN HEME APPEARS TO BE IN 450 00:17:51,200 --> 00:17:52,840 EXCESS OF GLOBEIN, HERE, THERE'S 451 00:17:52,840 --> 00:17:57,720 MORE HAD IN THE SAMPLE AND MORE 452 00:17:57,720 --> 00:17:58,000 APOPTOSIS. 453 00:17:58,000 --> 00:18:00,000 FOR THOSE SAIMENT EXPERIMENT 454 00:18:00,000 --> 00:18:03,720 WITH THE 5 HUGH, SAME RESULT, 455 00:18:03,720 --> 00:18:08,080 SLOW GLOBEIN PRODUCTION, DAY 7, 456 00:18:08,080 --> 00:18:13,800 INCREASE ROS, INCREASE 457 00:18:13,800 --> 00:18:14,040 APOPTOSIS. 458 00:18:14,040 --> 00:18:15,480 SO WE WENT ON AND SHOWED THAT 459 00:18:15,480 --> 00:18:17,560 HEME WAS CONTRIBUTING TO THIS BY 460 00:18:17,560 --> 00:18:20,960 TREATING THE CULTURE WITH THE 461 00:18:20,960 --> 00:18:23,080 ACETONE, WHICH INHIBITS THE 462 00:18:23,080 --> 00:18:25,600 SECOND STEP OF HEME SYNTHESIS 463 00:18:25,600 --> 00:18:27,800 AND SO THAT IN NORMAL, THERE'S A 464 00:18:27,800 --> 00:18:29,680 SLIGHT DECREASE IN THE NUMBER OF 465 00:18:29,680 --> 00:18:33,560 CELLS THAT ARE GENERATED BUT 466 00:18:33,560 --> 00:18:34,840 THERE THERE'S A DOUBLING OF 467 00:18:34,840 --> 00:18:37,600 CELLS YOU GET OUT OF DBA AND MDS 468 00:18:37,600 --> 00:18:42,960 CULTURES AND THE PRESENCE EVER 469 00:18:42,960 --> 00:18:44,040 SUCCINYLACETONE, SO THAT IT'S 470 00:18:44,040 --> 00:18:45,920 MORE CONSIST WENT THE AMOUNT OF 471 00:18:45,920 --> 00:18:51,160 GLOBEIN PRODUCED IN THESE 472 00:18:51,160 --> 00:18:51,600 CIRCUMSTANCES. 473 00:18:51,600 --> 00:18:53,480 SO FOR THE FIRST PART OF MY 474 00:18:53,480 --> 00:18:56,920 TALK, OUR DATA HAS SHOWN THAT 475 00:18:56,920 --> 00:18:59,280 THE CFE HEME, EXCEEDS USE IN 476 00:18:59,280 --> 00:19:02,520 HEMOGLOBIN OR EXPORT IN FLVCR 477 00:19:02,520 --> 00:19:05,280 AND BY EXTENSION, LIMITING HEME 478 00:19:05,280 --> 00:19:06,000 SYNTHESIS OR RESTRICTING IRON 479 00:19:06,000 --> 00:19:10,720 COULD BE A NOVEL WAY TO TREAT 480 00:19:10,720 --> 00:19:13,960 DBA AND DEL5 QSMAZE MDS AND 481 00:19:13,960 --> 00:19:16,200 SINCE DIVERSE MUTATIONS COULD 482 00:19:16,200 --> 00:19:18,960 IMPEE BRISK PROTEIN 483 00:19:18,960 --> 00:19:21,000 TRANSCRIPTION OR TRANSLATION, 484 00:19:21,000 --> 00:19:24,560 THIS MIGHT ADEQUATELY AFFECT THE 485 00:19:24,560 --> 00:19:27,800 PATIENTS WITHOUT DEL5 Q, BECAUSE 486 00:19:27,800 --> 00:19:33,040 THERE'S IBDICATIONS IN M, D--MDS 487 00:19:33,040 --> 00:19:35,480 PATIENTS THAT COULD HELP MORE 488 00:19:35,480 --> 00:19:38,760 CRITICALLY THAT HEME SYNTHESIS. 489 00:19:38,760 --> 00:19:39,800 SO WE CONVERTED THESE STUDIES TO 490 00:19:39,800 --> 00:19:41,440 LOOK AT A MOUSE MODEL TO GET 491 00:19:41,440 --> 00:19:43,080 MORE INSIGHT TO WHAT ACTUALLY 492 00:19:43,080 --> 00:19:48,440 HEME IS DOING AS RED CELLS 493 00:19:48,440 --> 00:19:48,680 MATURE. 494 00:19:48,680 --> 00:19:50,800 SO FLVCR HAVE MICE, HAVE ANEMIA 495 00:19:50,800 --> 00:19:53,200 THAT'S SIMILAR TO DBA AND 5 Q 496 00:19:53,200 --> 00:20:01,000 MDS PATIENTS AND THERE IS A LOW 497 00:20:01,000 --> 00:20:02,720 [INDISCERNIBLE], LOW HEMOGLOBIN 498 00:20:02,720 --> 00:20:04,480 AND HIGH DEL5 Q, AND SO CELLS 499 00:20:04,480 --> 00:20:06,000 DIE AT THE SAME STAGE BETWEEN 500 00:20:06,000 --> 00:20:08,800 THE EARLIER STAGES OF 501 00:20:08,800 --> 00:20:10,040 DIFFERENTIATION AS SHOWN HERE. 502 00:20:10,040 --> 00:20:13,400 AT THE TIME THERE CFUE OR 503 00:20:13,400 --> 00:20:14,480 PROERYTHROPOIESIS AND THESE 504 00:20:14,480 --> 00:20:15,480 EARLY CELLS HAVE HEME CONTHAT HE 505 00:20:15,480 --> 00:20:18,320 WENT'S HIGH AND A HIGH SET OF 506 00:20:18,320 --> 00:20:20,200 PLASMIC ROS, SO A MODEL THAT'S 507 00:20:20,200 --> 00:20:21,520 VERY SIMILAR AND WHAT'S ACTUALLY 508 00:20:21,520 --> 00:20:24,120 REALLY COOL ABOUT THIS MODEL IS 509 00:20:24,120 --> 00:20:25,720 JUST NOTHING ELSE WRONG WITH 510 00:20:25,720 --> 00:20:27,000 THESE CELLS, IN OTHER WORDS 511 00:20:27,000 --> 00:20:28,040 WHITE CELL PRODUCTION IS FINE, 512 00:20:28,040 --> 00:20:30,440 EVERYTHING IS FINE ABOUT THEM, 513 00:20:30,440 --> 00:20:34,800 DIFFERENTIATION OTHERWISE IS 514 00:20:34,800 --> 00:20:36,200 FINE, BUT THEY CAN'T CONTROL 515 00:20:36,200 --> 00:20:38,240 HEME WHEN CELLS GET TO THE STAGE 516 00:20:38,240 --> 00:20:45,160 OF CFUE, SO THEREFORE WE CAN ASK 517 00:20:45,160 --> 00:20:45,800 REALLY WHAT IS HEME DOING, THAT 518 00:20:45,800 --> 00:20:46,400 MAKES WHAT WOULD BE A NORMAL 519 00:20:46,400 --> 00:20:46,920 CELL NO LONGER ABLE TO 520 00:20:46,920 --> 00:20:48,840 DIFFERENTIATE AND TO DIE LEADING 521 00:20:48,840 --> 00:20:49,760 TO INEFFECTIVE ERYTHROPOIESIS. 522 00:20:49,760 --> 00:20:51,600 SO HERE IS EXACTLY THE SAME 523 00:20:51,600 --> 00:20:54,240 SLIDE I SHOWED BEFORE, STARTING 524 00:20:54,240 --> 00:20:56,680 THE PROCESS, THE RECEPTOR GOING 525 00:20:56,680 --> 00:20:59,800 UP, TURNING ON THE RECEPTOR, 526 00:20:59,800 --> 00:21:01,880 CD71, NOW THEREYA A MARKER IN 527 00:21:01,880 --> 00:21:05,400 MOUSE FOR THE SAME PROTEINS 528 00:21:05,400 --> 00:21:07,480 WHICH IS TER119, WITH WITH WHEN 529 00:21:07,480 --> 00:21:09,160 THE IRON ENTERS, THE THEY ARE 530 00:21:09,160 --> 00:21:11,240 APIECED AND BINDS AND INDUCED 531 00:21:11,240 --> 00:21:13,200 TRANSLATION, THAT STARTS HEME 532 00:21:13,200 --> 00:21:15,000 SYNTHESIS, HEME SYNTHESIS 533 00:21:15,000 --> 00:21:16,240 INDUCES GLOBEIN AND 534 00:21:16,240 --> 00:21:19,920 TRANSCRIPTION AND TRANSLATION BY 535 00:21:19,920 --> 00:21:21,320 INHIBITING RECEPTORS AND IT 536 00:21:21,320 --> 00:21:27,320 INITIATES AND THERE'S TIME WHERE 537 00:21:27,320 --> 00:21:28,960 IT EXCEEDS GLOBEIN AND WHEN THEY 538 00:21:28,960 --> 00:21:30,440 ARE RELEASED THE CELLS DIE. 539 00:21:30,440 --> 00:21:31,800 SO WE DID SINGLE CELL 540 00:21:31,800 --> 00:21:33,200 EXPERIMENTS BECAUSE WHAT WE WANT 541 00:21:33,200 --> 00:21:37,760 TO IS AN INDIVIDUAL CELL REACHES 542 00:21:37,760 --> 00:21:44,000 THIS OBSTACLE FOR ITSELF HAS TO 543 00:21:44,000 --> 00:21:45,400 DO WITH HEME EXACTLY WHEN'S 544 00:21:45,400 --> 00:21:46,880 GOING ON AT THE SINGLE CELL 545 00:21:46,880 --> 00:21:48,080 LEVEL, NOT AT THE POPULATION 546 00:21:48,080 --> 00:21:49,960 LEVEL BUT THE INDIVIDUAL CELL 547 00:21:49,960 --> 00:21:51,200 THAT HITS OBSTACLES, NEEDS TO 548 00:21:51,200 --> 00:21:52,520 OVERCOME THEM IN ORDER TO BE A 549 00:21:52,520 --> 00:21:55,960 SUCCESSFUL IN THE 550 00:21:55,960 --> 00:21:56,760 DIFFERENTIATION SEQUENCE. 551 00:21:56,760 --> 00:22:00,720 AND SO WE IDENTIFIED 552 00:22:00,720 --> 00:22:01,440 IDENTIFIERINGS THROUGH FLOW 553 00:22:01,440 --> 00:22:04,000 CYTOMETRY AND ISOLATED SIGNAL 554 00:22:04,000 --> 00:22:10,120 CELLS AND QUANTITATED THE CELL 555 00:22:10,120 --> 00:22:12,200 SURFACE MARKERS, AND LINK THE 556 00:22:12,200 --> 00:22:14,240 PROTEIN EXPRESSION OF A SINGLE 557 00:22:14,240 --> 00:22:17,000 CELL TO HIS RNASEQ. 558 00:22:17,000 --> 00:22:20,760 FIRST I JUST SHOWED THE METHODS 559 00:22:20,760 --> 00:22:22,000 RELIABLE. 560 00:22:22,000 --> 00:22:23,840 HEMOGLOBIN, BETTER CHAIN, ALPHA 561 00:22:23,840 --> 00:22:25,280 CHAIN COME FROM DIFFERENT 562 00:22:25,280 --> 00:22:26,720 CHROMOSOMES BUT THEY'RE 563 00:22:26,720 --> 00:22:28,000 EXCLUSIVELY CO REGULATED SO WE 564 00:22:28,000 --> 00:22:29,320 WANT IN ANY SINGLE CELL ANALYSIS 565 00:22:29,320 --> 00:22:31,800 TO HAVE AN ABSOLUTE CORRELATION 566 00:22:31,800 --> 00:22:32,840 BETWEEN HEMOGLOBIN BETA CHAIN 567 00:22:32,840 --> 00:22:35,000 AND ALPHA CHAIN AND THIS IS AN 568 00:22:35,000 --> 00:22:35,760 MAIRKSZING CORRELATION ACROSS 569 00:22:35,760 --> 00:22:42,880 OUR NORMAL AND OUR ABNORMAL 570 00:22:42,880 --> 00:22:43,200 MICE. 571 00:22:43,200 --> 00:22:49,280 ON THE OTHER HAND THE PROTEIN, 572 00:22:49,280 --> 00:23:00,000 TER119, WITH THE MESSAGE WITH 573 00:23:00,000 --> 00:23:01,320 THE GLYCA, CORRELATION, SO 574 00:23:01,320 --> 00:23:03,480 THERE'S NOT A PERFECT 575 00:23:03,480 --> 00:23:04,680 CORRELATION BETWEEN MESSAGE AND 576 00:23:04,680 --> 00:23:05,600 PROTEIN EXPRESSION AND THE SAME 577 00:23:05,600 --> 00:23:09,320 THING CAN BE SEPARATE, CD71 AND 578 00:23:09,320 --> 00:23:10,760 THE GENE TRANSFER RECEPTOR THAT 579 00:23:10,760 --> 00:23:11,960 IS WELL CORRELATED SO THIS 580 00:23:11,960 --> 00:23:13,080 LOOKED PRETTY GOOD AND LOOKED 581 00:23:13,080 --> 00:23:14,320 LIKE WE EXPECTED THE METHODS IN 582 00:23:14,320 --> 00:23:18,800 THE SINGLE CELL LEVEL WERE 583 00:23:18,800 --> 00:23:20,800 ACTUALLY REPORTING OUT FROM WHAT 584 00:23:20,800 --> 00:23:21,760 1 WOULD ANTICIPATE. 585 00:23:21,760 --> 00:23:24,200 SO WHEN PEOPLE GENTLEMENLY DO 586 00:23:24,200 --> 00:23:28,720 EXPERIMENTS AND LOOK AT 587 00:23:28,720 --> 00:23:29,960 ERYTHROPOIESIS DIFFERENTIATION 588 00:23:29,960 --> 00:23:32,160 USING TER119 AS A PROMOTER, THEY 589 00:23:32,160 --> 00:23:34,600 CALL IT TERLOW OR HIGH AND MAKE 590 00:23:34,600 --> 00:23:36,320 IT THE BIENINARY DIFFERENTIATION 591 00:23:36,320 --> 00:23:38,560 BETWEEN A MATURE AND LESS MATURE 592 00:23:38,560 --> 00:23:40,560 CELL BUT INSEDENTARY WE LOOKED 593 00:23:40,560 --> 00:23:43,600 AT UNBIAS ANALYSIS AND LOOKED AT 594 00:23:43,600 --> 00:23:45,200 WHERE CELLS DISTRIBUTED BASED ON 595 00:23:45,200 --> 00:23:47,400 THE INTENSITY OF THE TER119 596 00:23:47,400 --> 00:23:49,720 EXPRESSION AND WHEN YOU DO THAT 597 00:23:49,720 --> 00:23:50,880 IT DIVIDES INTO 4 DIFFERENT 598 00:23:50,880 --> 00:23:53,200 STAGES WHICH WE CALL ABCD AND 599 00:23:53,200 --> 00:23:54,200 WHAT BECAME IMPORTANT, THIS 600 00:23:54,200 --> 00:23:56,840 ALLOWED US TO LOOK AT THE VERY 601 00:23:56,840 --> 00:23:58,640 EARLY EVENTS, STAGE A WITHOUT 602 00:23:58,640 --> 00:24:00,160 BEING IMPACTED BY WHAT HAPPENS 603 00:24:00,160 --> 00:24:02,200 LATER, IF YOU'RE GOING TO MAKE 604 00:24:02,200 --> 00:24:04,040 AN ARBITRARY CUT AND INCLUDE ALL 605 00:24:04,040 --> 00:24:07,160 THOSE CELLS IN YOUR ANALYSIS. 606 00:24:07,160 --> 00:24:09,400 AND SO WHEN 1 LOOKS AT NORMAL 607 00:24:09,400 --> 00:24:13,120 MICE, SEPARATE A, B, C, D, BASED 608 00:24:13,120 --> 00:24:14,040 ON THIS UNBIASED CUT OFF AND 609 00:24:14,040 --> 00:24:16,000 THEN WE USE THE IDENTICAL CUT 610 00:24:16,000 --> 00:24:19,400 OFFS IN OUR DELETED MICE, A, B, 611 00:24:19,400 --> 00:24:20,600 C, D, AND THE FIRST AND VERY 612 00:24:20,600 --> 00:24:22,200 NICE THING WE FOUND IS THAT 613 00:24:22,200 --> 00:24:23,600 AMONG THE A-CELL, THE VERY 614 00:24:23,600 --> 00:24:26,240 EARLIER CELLS, HALF THE CELLS 615 00:24:26,240 --> 00:24:27,560 WERE DYING OR DEAD. 616 00:24:27,560 --> 00:24:31,200 SO IT CONFIRMED THAT INDEED, 617 00:24:31,200 --> 00:24:33,240 DEATH WAS OCCURRING AT THIS 618 00:24:33,240 --> 00:24:38,200 EARLY STAGE, AT THE STAGE OF 619 00:24:38,200 --> 00:24:38,880 ERYTHROPOIESIS DIFFERENTIATION. 620 00:24:38,880 --> 00:24:41,720 WE THEN WERE ABLE TO SAY, WELL 621 00:24:41,720 --> 00:24:43,400 WHAT HAPPENS TO THESE CELLS AT A 622 00:24:43,400 --> 00:24:44,920 SINGLE CELL LEVEL ACROSS THE 623 00:24:44,920 --> 00:24:45,840 DIFFERENTIATION SEQUENCE AND 624 00:24:45,840 --> 00:24:47,480 THERE WERE REALLY SURPRISING 625 00:24:47,480 --> 00:24:47,720 RESULTS. 626 00:24:47,720 --> 00:24:52,800 AND I WILL SHOW YOU A FEW OF 627 00:24:52,800 --> 00:24:54,480 THOSE. 628 00:24:54,480 --> 00:24:57,920 SO HERE IS A PLOT OF THE 629 00:24:57,920 --> 00:24:59,000 EXPRESSION OF RIBOSOMAL 630 00:24:59,000 --> 00:25:02,600 PROTEINS, WHICH WAS THE MOST 631 00:25:02,600 --> 00:25:05,840 DIFFERENTIALLY EXPRESSED BETWEEN 632 00:25:05,840 --> 00:25:06,960 THE FLVCR-DELETED AND THE 633 00:25:06,960 --> 00:25:07,280 CONTROL MICE. 634 00:25:07,280 --> 00:25:11,160 AND IN THE EARLY STAGES A AND B, 635 00:25:11,160 --> 00:25:14,040 THE EXCESS AMOUNTS OF THE RIKE O 636 00:25:14,040 --> 00:25:15,760 STUDIES OF MULTIPLE ENDOCRINAL 637 00:25:15,760 --> 00:25:17,200 CONTENTS BEING DELETED IN THE 638 00:25:17,200 --> 00:25:17,480 MOUSE. 639 00:25:17,480 --> 00:25:19,000 WE THOUGHT THIS IS CRAZY. 640 00:25:19,000 --> 00:25:20,840 WHAT DOES THIS MEAN IN BUT 641 00:25:20,840 --> 00:25:23,000 REMEMBER THIS HEME EXCESS, 642 00:25:23,000 --> 00:25:24,000 YOU'RE MAKING, NORMALLY THIS IS 643 00:25:24,000 --> 00:25:26,200 HEME, YOU'RE MAKING GLOBEIN, 644 00:25:26,200 --> 00:25:27,080 HEME'S MAKING GLOBEIN HAPPEN, 645 00:25:27,080 --> 00:25:29,720 HEME MUST BE MAKING RNA PROTEINS 646 00:25:29,720 --> 00:25:32,000 BEING MADE SO THERE ARE ACTUALLY 647 00:25:32,000 --> 00:25:34,080 RIBOSOMES TO TRANSLATE THE HEME, 648 00:25:34,080 --> 00:25:35,480 TO TRANSLATE THE HEME MESSAGE 649 00:25:35,480 --> 00:25:37,680 THAT YOU NEED TO MAKE TO 650 00:25:37,680 --> 00:25:38,280 TRANSLATE THE GLOBEIN MESSAGE 651 00:25:38,280 --> 00:25:39,800 THAT YOU NEED TO MAKE GLOBEIN 652 00:25:39,800 --> 00:25:41,960 AND IT MAKES SENSE THAT HEME 653 00:25:41,960 --> 00:25:43,600 BESIDES DURPING ON GLOBEIN 654 00:25:43,600 --> 00:25:44,280 TRANSCRIPTION AND TRANSLATION IS 655 00:25:44,280 --> 00:25:46,480 TURNING ON THE REST OF A 656 00:25:46,480 --> 00:25:48,200 PROCESSES THAT ARE NECESSARY TO 657 00:25:48,200 --> 00:25:49,400 MAKE QUICK GLOBEIN TO HAVE IT 658 00:25:49,400 --> 00:25:51,880 MATCH THE HEME THAT IS PRESENT. 659 00:25:51,880 --> 00:25:53,800 BUT WE WANTED TO TEST THIS 660 00:25:53,800 --> 00:25:54,200 DIRECTLY. 661 00:25:54,200 --> 00:26:03,600 SO WE TOOK HUMAN CELLS, THAT ARE 662 00:26:03,600 --> 00:26:05,920 CD36 POSITIVE, GLYCO HEME AND 663 00:26:05,920 --> 00:26:07,600 TIMES THEM BY 0 WHICH GAVE 664 00:26:07,600 --> 00:26:11,400 THEMA, AND BYPASS--WHICH IS THE 665 00:26:11,400 --> 00:26:12,440 RATE OF SYNTHESIS. 666 00:26:12,440 --> 00:26:13,880 SO TIMES 0, YOU BYPASS, THE 667 00:26:13,880 --> 00:26:15,720 STEP, AND YOU IMMEDIATELY START 668 00:26:15,720 --> 00:26:17,440 HEME SYNTHESIS AND THEN SAID 669 00:26:17,440 --> 00:26:21,160 WHAT HAPPENS 15 MINUTES LATER? 670 00:26:21,160 --> 00:26:22,400 FIFTEEN MINUTES AFTER HEME 671 00:26:22,400 --> 00:26:27,360 SYNTHESIS IS INDUCED IN ERYTH 672 00:26:27,360 --> 00:26:30,720 ROADWAY CELLS AND MOST OF THEM 673 00:26:30,720 --> 00:26:32,840 GO UP, THEY GO INITIALLY AND GO 674 00:26:32,840 --> 00:26:33,760 UP BY PROTEIN TO FOLLOW. 675 00:26:33,760 --> 00:26:38,240 SO THIS,A PEERED TO BE A VERY 676 00:26:38,240 --> 00:26:39,120 PHYSIOLOGIC PROCESS IN NORMAL 677 00:26:39,120 --> 00:26:41,000 CELLS AND WE CAN CONFIRM THAT AS 678 00:26:41,000 --> 00:26:43,000 WELL BY LOOKING AT RIBOSOMAL RNA 679 00:26:43,000 --> 00:26:44,840 WHICH IS A SURROGATE FOR THE 680 00:26:44,840 --> 00:26:45,480 AMOUNT OF RIEB SO STUDIES OF 681 00:26:45,480 --> 00:26:46,800 MULTIPLE ENDOCRINES THAT ARE 682 00:26:46,800 --> 00:26:49,160 PRESENT AND WITHIN 30 MINUTES OF 683 00:26:49,160 --> 00:26:51,600 ADDING ALA, THERE WAS EXTRA 684 00:26:51,600 --> 00:26:53,880 RIBOSOMAL RNA OR EXTRA RIBOSOMES 685 00:26:53,880 --> 00:27:00,880 PRESENT IN THESE EARLY CELLS. 686 00:27:00,880 --> 00:27:02,520 A SECOND SORT OF SURPRISE IS 687 00:27:02,520 --> 00:27:08,000 THAT HEME TURNS OFF GATAC1. 688 00:27:08,000 --> 00:27:09,760 SO HERE'S GATO1 AND THE SAME 689 00:27:09,760 --> 00:27:15,600 AMOUNT OR LESSER CELLS, A 690 00:27:15,600 --> 00:27:24,120 CLUSTER, A GATAC1--THE GATAC1 691 00:27:24,120 --> 00:27:26,120 TARGET AND EXPRESSION GOES UP AS 692 00:27:26,120 --> 00:27:29,000 DIFFERENTIATION HAPPENS BUT HERE 693 00:27:29,000 --> 00:27:29,560 IT DOESN'T. 694 00:27:29,560 --> 00:27:31,880 THIS IS SHOWN HERE BETTER IN THE 695 00:27:31,880 --> 00:27:33,520 CLUSTER EXPRESSION ANALYSIS, AND 696 00:27:33,520 --> 00:27:36,840 A, MORE OR LESS THE SAME, BUT IN 697 00:27:36,840 --> 00:27:40,360 B, C, D, THE LATER STAGES HIGH 698 00:27:40,360 --> 00:27:41,680 EXPRESSION OF TARGETS, WHILE 699 00:27:41,680 --> 00:27:43,320 THERE ISN'T HIGH EXPRESSION AND 700 00:27:43,320 --> 00:27:48,360 THAT THAT IS HEME EXCESSED FOR 701 00:27:48,360 --> 00:27:49,960 FLVCR DELETED AND IF WE CAN SHOW 702 00:27:49,960 --> 00:27:53,400 THAT AGAIN BY GOING TO THE LATER 703 00:27:53,400 --> 00:27:57,000 CELLS, THE FOREIGN A AND CD36 704 00:27:57,000 --> 00:28:00,280 POSITIVE CELLS SO THE MIDDLE OF 705 00:28:00,280 --> 00:28:02,480 THE CELL DIFFERENTIATION AND 706 00:28:02,480 --> 00:28:08,080 WITHIN 15 MINUTES OF ADDING ALA 707 00:28:08,080 --> 00:28:09,400 TO THE CD36-GLYA, IN THE CELLS 708 00:28:09,400 --> 00:28:15,480 THERE IS A DECREASE OF MRNA IN 709 00:28:15,480 --> 00:28:16,200 GATAC1. 710 00:28:16,200 --> 00:28:18,280 ONE MORE METHOD, WE WENT TO K526 711 00:28:18,280 --> 00:28:20,800 CELLS WHICH IS A CELL LINE 712 00:28:20,800 --> 00:28:24,320 THAT'S ERYTH ROADWAY IN ITS 713 00:28:24,320 --> 00:28:26,240 EXPRESSION AND 5562 CELLS AT 714 00:28:26,240 --> 00:28:33,920 TIME 0 HAVE RELATIVELY 1 IN THIS 715 00:28:33,920 --> 00:28:36,160 CONTENT, OF HEME CONTENT AND THE 716 00:28:36,160 --> 00:28:37,920 AMOUNT OF HEME CONTENT AFTER 717 00:28:37,920 --> 00:28:40,200 ADDING ALA IS HIGH, 15 MINUTES 718 00:28:40,200 --> 00:28:40,440 LATER. 719 00:28:40,440 --> 00:28:42,920 HOWEVER, IF YOU ADD ALA WHICH 720 00:28:42,920 --> 00:28:44,880 BYPASSES THE FIRST STEP, BUT 721 00:28:44,880 --> 00:28:46,360 THEN BLOCKS THE SECOND STEP, 722 00:28:46,360 --> 00:28:47,200 THIS DOESN'T HAPPEN. 723 00:28:47,200 --> 00:28:49,320 THERE'S NO INCREASE OF HEME 724 00:28:49,320 --> 00:28:49,800 CONTENT. 725 00:28:49,800 --> 00:28:52,800 AND SIMILARLY, IF 1 ADDING ALA, 726 00:28:52,800 --> 00:28:56,240 LOTS OF ROS, BUT IF YOU BLOCK 727 00:28:56,240 --> 00:28:59,320 HEME SYNTHESIS IMMEDIATELY AFTER 728 00:28:59,320 --> 00:29:01,520 STARTING IT, WITH THE ACETONE 729 00:29:01,520 --> 00:29:02,920 BEING ADDED AT THE SAME TIME 730 00:29:02,920 --> 00:29:05,240 THERE, 'S NO INCREASE IN ROS, 731 00:29:05,240 --> 00:29:06,560 AND THEREFORE THE SAME THING IS 732 00:29:06,560 --> 00:29:08,320 TRUE WITH HEME PROTEIN, GOES 733 00:29:08,320 --> 00:29:10,200 DOWN WITH ALA AND DOESN'T CHANGE 734 00:29:10,200 --> 00:29:12,360 WITH ALA IS ACETONE AND GLOBEIN 735 00:29:12,360 --> 00:29:14,120 AND OTHER PROTEIN TARPGET OF 736 00:29:14,120 --> 00:29:17,200 COURSE HEME GOES UP WITH ALA AND 737 00:29:17,200 --> 00:29:19,880 GOES DOWN OR DOESN'T CHANGE WITH 738 00:29:19,880 --> 00:29:22,440 ALA AND THE ACETONE. 739 00:29:22,440 --> 00:29:28,480 SO A NICE CONCEPT OF CAUSE AND 740 00:29:28,480 --> 00:29:30,320 EFFECT BEING SHOWN HERE THAT 741 00:29:30,320 --> 00:29:31,400 HEME IS INDUCING THESE CHANGES 742 00:29:31,400 --> 00:29:35,320 THAT WE'VE BEEN OBSERVING IN THE 743 00:29:35,320 --> 00:29:36,920 PRIOR EXPERIMENTATION. 744 00:29:36,920 --> 00:29:39,760 SO WHAT DOES THIS MEAN? 745 00:29:39,760 --> 00:29:43,040 SORT OF IN A SUMMARY, THAT THIS 746 00:29:43,040 --> 00:29:47,680 IS HOW I THINK ABOUT RED CELL 747 00:29:47,680 --> 00:29:49,840 DIFFERENTIATION. 748 00:29:49,840 --> 00:29:51,520 THAT GATAC1 UPREGULATES ALAS 749 00:29:51,520 --> 00:29:53,240 TRANSCRIPTION, IT STARTS THE 750 00:29:53,240 --> 00:29:56,400 PROCESS, HEME PROCESS STARTS AND 751 00:29:56,400 --> 00:30:02,800 UPREGULATES GLOBEIN SYNTH TIS 752 00:30:02,800 --> 00:30:05,680 AND AT THIS STAGE A BUSY TIME OF 753 00:30:05,680 --> 00:30:07,880 ERYTHROPOIESIS WHILE THIS IS 754 00:30:07,880 --> 00:30:08,800 HAPPENING. 755 00:30:08,800 --> 00:30:11,440 THE CELLS ARE DETECTED FROM THE 756 00:30:11,440 --> 00:30:13,040 COMA HEME AND THE GLOBEIN 757 00:30:13,040 --> 00:30:14,200 DISCONNECT BECAUSE OF THE 758 00:30:14,200 --> 00:30:16,560 PROCESS HERE THAT EXPORTS THAT 759 00:30:16,560 --> 00:30:16,800 HEME. 760 00:30:16,800 --> 00:30:18,440 BUT THEN UNDER NORMAL 761 00:30:18,440 --> 00:30:22,240 CIRCUMSTANCES, THIS RED CELLS 762 00:30:22,240 --> 00:30:24,320 MATURE, THE FLVCR CELL SURFACE 763 00:30:24,320 --> 00:30:26,200 EXPRESSION ACTUALLY DECREASES, 764 00:30:26,200 --> 00:30:28,160 IT'S ONLY 60ICIZEED AT STAGE AND 765 00:30:28,160 --> 00:30:30,200 NO RNA PRESENT AT ANY STAGE 766 00:30:30,200 --> 00:30:31,800 AFTER THAT AND WITH CELL 767 00:30:31,800 --> 00:30:33,560 DIVISION, THE AMOUNT OF THE CELL 768 00:30:33,560 --> 00:30:35,600 SURFACE DECREASES AND SO THE 769 00:30:35,600 --> 00:30:39,120 LATER CELLS ARE MORE SENSITIVE 770 00:30:39,120 --> 00:30:41,880 TO EXCESS HEME AND THIS IS 771 00:30:41,880 --> 00:30:44,000 ACTUALLY DOWN REGULATED AND 772 00:30:44,000 --> 00:30:45,480 ERYTHROPOIESIS CAN TORMINATE 773 00:30:45,480 --> 00:30:45,760 NORMALLY. 774 00:30:45,760 --> 00:30:48,240 SO THERE'S ACTUALLY A GATAC1 CO 775 00:30:48,240 --> 00:30:51,960 REGULATION IN FEEDBACK LOOP THAT 776 00:30:51,960 --> 00:30:53,200 ALLOWS NORMAL DIFFERENTIATION 777 00:30:53,200 --> 00:30:54,480 WITH THIS KINETIC VERY FAST 778 00:30:54,480 --> 00:30:57,960 PROCESS TO BE SO STRICTLY 779 00:30:57,960 --> 00:30:58,480 CONTROLLED. 780 00:30:58,480 --> 00:31:00,800 AND WHAT HAPPENS WHEN HEME 781 00:31:00,800 --> 00:31:02,920 ACCUMULATES EARLY BECAUSE OF 782 00:31:02,920 --> 00:31:06,440 ABNORMALITIES THAT ARE PRESENT 783 00:31:06,440 --> 00:31:13,800 IN DBA AND IN THE POETICESIS, IT 784 00:31:13,800 --> 00:31:17,920 PRACTICE MATURELY DOWN REGULATES 785 00:31:17,920 --> 00:31:19,800 GATAC1 AND ERYTH ROUGH ATOM 786 00:31:19,800 --> 00:31:23,040 PORESIS TERMINATES THE EFFECTIVE 787 00:31:23,040 --> 00:31:23,400 POETICESIS. 788 00:31:23,400 --> 00:31:28,640 SO 2, GENETIC MUTATIONS IN THIS, 789 00:31:28,640 --> 00:31:30,600 RESULTS IN SLOW TRANSCRIPTION OR 790 00:31:30,600 --> 00:31:34,320 TRANSLATION AND DBA TRANSLATION 791 00:31:34,320 --> 00:31:37,280 IS SLOWED BECAUSE THIS IS A 792 00:31:37,280 --> 00:31:39,400 RIBOSOMAL ABNORMALITY AND HEME 793 00:31:39,400 --> 00:31:41,080 EXCLUDES GLOBEIN. 794 00:31:41,080 --> 00:31:43,160 THE EXCESS HEME CAUSES INCREASED 795 00:31:43,160 --> 00:31:44,800 ROS IN CELL DEATH, IT INCREASES 796 00:31:44,800 --> 00:31:45,760 RIBOSOMAL PROTEINS O STUDIES OF 797 00:31:45,760 --> 00:31:49,200 MULTIPLE ENDOCRINAL PROTEIN 798 00:31:49,200 --> 00:31:51,640 SIPGHT SIS, BUT AMPLIFY ANY 799 00:31:51,640 --> 00:31:56,000 PROTEIN BALANCE AT PRESENT AND 800 00:31:56,000 --> 00:31:57,720 DECREASES TRANSCRIPTION OF GENES 801 00:31:57,720 --> 00:32:00,680 AND MITOSIS, I WILL SHOW THIS IN 802 00:32:00,680 --> 00:32:02,760 A MINUTE CAUSING SIGNIFYITOSEIS 803 00:32:02,760 --> 00:32:06,520 AND RESULTS IN THE EARLY AND 804 00:32:06,520 --> 00:32:07,120 PREMATURE DIFFERENTIATION 805 00:32:07,120 --> 00:32:08,840 TERMINATION PROGRAM. 806 00:32:08,840 --> 00:32:10,960 SO TOGETHER THESE MULTIPLE WAYS 807 00:32:10,960 --> 00:32:13,960 ASSURES THE CELLS DON'T MAKE IT 808 00:32:13,960 --> 00:32:17,000 IF THEY'RE ABNORMAL, IF THERE'S 809 00:32:17,000 --> 00:32:24,520 EXCESS HEME TO THE GLOBEIN AT 810 00:32:24,520 --> 00:32:25,040 THIS JUNCTURE. 811 00:32:25,040 --> 00:32:33,480 SO SHOWING SHOWING ANOTHER WAY, EXCESS 812 00:32:33,480 --> 00:32:40,640 HEME WILL LEAD TO AND TERMINAL 1 813 00:32:40,640 --> 00:32:45,400 AND IMPLYING AND BY IMPAIRING IT 814 00:32:45,400 --> 00:32:48,920 OR OUR WAY THAT 1 WOULD ACTUALLY 815 00:32:48,920 --> 00:32:51,400 IMPROVE THE PROCESS ALLOWING 816 00:32:51,400 --> 00:32:53,720 ABNORMAL CELLS TO DIFFERENTIATE 817 00:32:53,720 --> 00:32:54,920 NOR COMPLETELY, AND WHAT'S 818 00:32:54,920 --> 00:32:55,760 INCREEINGING ABOUT THIS IDEA AS 819 00:32:55,760 --> 00:32:58,800 A WAY TO GO IN TERMS OF THAT 820 00:32:58,800 --> 00:33:00,320 THERAPEUTIC TARGET, IS THAT ALL 821 00:33:00,320 --> 00:33:06,120 THE THERAPIES THAT HAVE BEEN 822 00:33:06,120 --> 00:33:08,000 APPROVED OR STUDIED SO FAR IS A 823 00:33:08,000 --> 00:33:11,440 NUMBER OF THESE CELLS AND SO 824 00:33:11,440 --> 00:33:16,760 THIS WOULD INCLUDE PREDINISONE, 825 00:33:16,760 --> 00:33:18,120 MDS AND OTHER HYPER METHALATING 826 00:33:18,120 --> 00:33:18,800 AREAS IN THE CLINICAL TRIALS 827 00:33:18,800 --> 00:33:21,600 THAT ARE UNDER WAY, AND SO IF 828 00:33:21,600 --> 00:33:25,800 THERE WERE TO BE ERYTHROID CELLS 829 00:33:25,800 --> 00:33:29,440 AND WHEN ADDED TO SOMETHING FOR 830 00:33:29,440 --> 00:33:30,640 GENERATEDIS, THESE SHOULD BE 831 00:33:30,640 --> 00:33:31,920 SINNER GESTIC AND THESE ARE 832 00:33:31,920 --> 00:33:34,120 AVENUES WE ARE PURSUING IN TERMS 833 00:33:34,120 --> 00:33:35,000 OF THE THERAPEUTIC EXTENSION OF 834 00:33:35,000 --> 00:33:39,200 SOME OF THESE BASIC INSIGHTS. 835 00:33:39,200 --> 00:33:43,200 SO TO CONTINUE ON A LITTLE BIT. 836 00:33:43,200 --> 00:33:45,560 THERE IS AN ANIMAL MODEL OF DBA, 837 00:33:45,560 --> 00:33:50,120 IT'S NOT A GREAT ANIMAL MODEL OF 838 00:33:50,120 --> 00:33:53,920 DBA, AND INDEED THE FINDINGS ARE 839 00:33:53,920 --> 00:33:56,040 SIMILAR TO FLVCR, AND CONFIRM A 840 00:33:56,040 --> 00:33:58,440 ROLE FOR HEME TOXICITIES INVIVO. 841 00:33:58,440 --> 00:33:59,840 AS WELL AS ADDITIONAL PATHWAYS 842 00:33:59,840 --> 00:34:03,240 THAT WE'RE ABLE TO PICK UP THAT 843 00:34:03,240 --> 00:34:04,360 PICKED UP ON P53 ACTIVATION 844 00:34:04,360 --> 00:34:07,600 WHICH IS ANOTHER PIECE OF THE 845 00:34:07,600 --> 00:34:10,520 DBA PHYSIOLOGY AND IN LARGE PART 846 00:34:10,520 --> 00:34:11,880 BECAUSE OF THE ABNORMAL 847 00:34:11,880 --> 00:34:16,840 RIBOSOMAL STRUCTURE AND HOW 848 00:34:16,840 --> 00:34:18,280 THAT--HOW INCOMPLETE RIBOSOMES 849 00:34:18,280 --> 00:34:19,840 AND EXCESS RIBOSOMA PROTEINS 850 00:34:19,840 --> 00:34:21,920 WHEN YOU'RE LACKING A PARTICULAR 851 00:34:21,920 --> 00:34:25,560 RIBOSOME COMPONENT CAN ACTIVATE 852 00:34:25,560 --> 00:34:25,920 A CELL. 853 00:34:25,920 --> 00:34:30,400 SO AS SHOWN HERE, THOUGH, 854 00:34:30,400 --> 00:34:34,360 CONTROLS AGAIN HAVE LITTLE 855 00:34:34,360 --> 00:34:38,600 RIBOSOME ACCESS IN A AND B, BUT 856 00:34:38,600 --> 00:34:42,400 FLVCR DELETED ANIMALS AND RPL11 857 00:34:42,400 --> 00:34:44,160 HETEROZYGOUS ANIMALS ALL HAVE 858 00:34:44,160 --> 00:34:46,120 EXTRA RIBOSOMAL PROTEIN GENES 859 00:34:46,120 --> 00:34:49,400 AND THE EARLIER STAGES OF 860 00:34:49,400 --> 00:34:49,760 DIFFERENTIATION. 861 00:34:49,760 --> 00:34:52,040 SO THEY HAVE HEME EXCESS MAKING 862 00:34:52,040 --> 00:34:55,600 MORE RIBOSOMES TO ALLOW FOR MORE 863 00:34:55,600 --> 00:34:59,320 GLOBEIN TO BE MADE. 864 00:34:59,320 --> 00:35:01,360 GATAC1 IS SIMILAR, AND THE VERY 865 00:35:01,360 --> 00:35:04,040 EARLIER STAGES OF EACH OF THESE 866 00:35:04,040 --> 00:35:06,280 MICE MODELS, BUT IN THE CONTROL, 867 00:35:06,280 --> 00:35:10,880 IT INCREASES AND GATA 1 868 00:35:10,880 --> 00:35:12,240 INCREASES TREMENDOUSLY IN BCD 869 00:35:12,240 --> 00:35:14,800 AND THAT DOESN'T HAPPEN IN THE 870 00:35:14,800 --> 00:35:20,480 FLVCR MOUSE OR IN THE 871 00:35:20,480 --> 00:35:22,440 HETEROZYGOTE MOUSE AND I SHOWED 872 00:35:22,440 --> 00:35:24,080 THAT INTERESTINGLY THERE IS A 873 00:35:24,080 --> 00:35:24,960 LATE DIFFERENTIATION OF THE 874 00:35:24,960 --> 00:35:30,120 DELETED MOUSE AND THE RPLE 875 00:35:30,120 --> 00:35:31,760 HETEROZYGOTE MOUSE FOR THE 876 00:35:31,760 --> 00:35:34,280 SPINDLE PATHWAY SO THERE APPEARS 877 00:35:34,280 --> 00:35:35,720 TO BE SOME IMPAIRMENT OF 878 00:35:35,720 --> 00:35:38,400 DIVISION WHICH MAY BE 1 OF THE 879 00:35:38,400 --> 00:35:39,160 PATHWAYS LEADING TO 880 00:35:39,160 --> 00:35:43,000 MACROSIGNIFYITOSEIS IN ALL OF 881 00:35:43,000 --> 00:35:56,400 THESE SITUATIONS. 882 00:35:56,400 --> 00:36:02,800 SO LASTLY, WITH A SOMEWHAT 883 00:36:02,800 --> 00:36:03,920 DIFFERENT STUDY IN SINGLE CELL 884 00:36:03,920 --> 00:36:04,400 MOUSE. 885 00:36:04,400 --> 00:36:06,560 SOPHISTICATEDY WE STARTED WITH 886 00:36:06,560 --> 00:36:08,200 LINEAGE NEGATIVE MARROW CELLS 887 00:36:08,200 --> 00:36:13,000 AND USED BAR CODING AND 888 00:36:13,000 --> 00:36:15,920 SIGHT-SEEN WITH ANTIBODIES WITH 889 00:36:15,920 --> 00:36:20,920 A LINK TO A GENETIC CODE IN THE 890 00:36:20,920 --> 00:36:23,000 SAME SITUATION AND WE QUANTITATE 891 00:36:23,000 --> 00:36:24,520 AGAIN CELL SURFACE PROTEINS AT 892 00:36:24,520 --> 00:36:27,880 THE SAME TIME AS WE LOOK AT 893 00:36:27,880 --> 00:36:36,840 TOTAL TRANSCRIPT OHMS AND USED 894 00:36:36,840 --> 00:36:39,080 TECHNOLOGY WITH 10 X GENOMICS. 895 00:36:39,080 --> 00:36:40,520 SO WE ANALYZED CELLS DIRECTLY 896 00:36:40,520 --> 00:36:44,960 AFTER 3 AND 6 DAYS OF 897 00:36:44,960 --> 00:36:45,800 ERYTHROIDEXPANSION CULTURE AND 898 00:36:45,800 --> 00:36:46,800 THIS GAVE US THE OPPORTUNITY TO 899 00:36:46,800 --> 00:36:48,040 LOOK AT ABNORMALITIES NOARMAL 900 00:36:48,040 --> 00:36:50,800 CELLS HOW THEY DIFFERENTIATED AS 901 00:36:50,800 --> 00:36:51,920 WELL BY KEEPING THEM ALIVE 902 00:36:51,920 --> 00:36:54,840 DURING A CULTURE DIFFERENTIATION 903 00:36:54,840 --> 00:36:55,080 TIMING. 904 00:36:55,080 --> 00:37:00,800 WE ASSESS TRANSCRIPT OHMS WITH 905 00:37:00,800 --> 00:37:01,440 PSEUDOTIME ANALYSIS WHICH IS A 906 00:37:01,440 --> 00:37:03,120 DIFFERENT WAY TO LOOK AT IT OVER 907 00:37:03,120 --> 00:37:05,120 TIME AND DO THIS WE NEEDED A 908 00:37:05,120 --> 00:37:07,720 NOVEL APPROACH CAN WHICH 909 00:37:07,720 --> 00:37:08,800 INCLUDED COMPLETELY PROCESSED 910 00:37:08,800 --> 00:37:11,600 MRNAs AND THESE ARE HIGHLY 911 00:37:11,600 --> 00:37:15,160 PREVALENT AND AND THIS ALLOWED 912 00:37:15,160 --> 00:37:17,600 US TO USE PSEUDOTIME AND ALIGN 913 00:37:17,600 --> 00:37:18,640 WITH DIFFERENT EXPERIMENTS AND 914 00:37:18,640 --> 00:37:22,200 THIS WE EARNED IS A LARGE NUMBER 915 00:37:22,200 --> 00:37:30,160 OF MRNAs WITH CHAIN INTRONS 916 00:37:30,160 --> 00:37:33,600 AND BESIDES--WITH ALL OF THESE 917 00:37:33,600 --> 00:37:34,680 INSIGHTS INTO REGULATORY 918 00:37:34,680 --> 00:37:36,160 DIFFERENTIATION. 919 00:37:36,160 --> 00:37:38,320 WHEN YOU TRACK BY PSEUDOTIME YOU 920 00:37:38,320 --> 00:37:41,080 LOOK AT THE PRESENCE OF A 921 00:37:41,080 --> 00:37:46,560 MESSAGE AND HOW IT PREDATES, HOW 922 00:37:46,560 --> 00:37:48,800 IT SPLICES THE MESSAGE AS A WAY 923 00:37:48,800 --> 00:37:51,600 OF ORYEBTS TIME AND FOR EVERY 924 00:37:51,600 --> 00:37:53,960 CELL LINEAGE, IT DOES A 925 00:37:53,960 --> 00:37:55,160 WOBDERFUL JOB BUT IN 926 00:37:55,160 --> 00:37:57,400 ERYTHROPOIESIS IT PREDICTS A 927 00:37:57,400 --> 00:37:59,280 BACKWARDS DIFFERENTIATION, WHERE 928 00:37:59,280 --> 00:38:00,040 YOU GET LATENT ERYTHROPOIESIS 929 00:38:00,040 --> 00:38:02,080 BECAUSE OF THE AMOUNT OF 930 00:38:02,080 --> 00:38:03,720 RETAINED INTRONS THAT OCCUR ON 931 00:38:03,720 --> 00:38:07,000 ROUTE AND COMMONLY OFFER ANOTHER 932 00:38:07,000 --> 00:38:08,480 INSIGHT TO THE REGULATIONS THAT 933 00:38:08,480 --> 00:38:21,600 OCCUR DURING THAT PROCESS. 934 00:38:21,600 --> 00:38:38,080 AND FIND THE CELL TYPES AS SHOWN 935 00:38:38,080 --> 00:38:40,840 HERE AND USING ANTIBODY AND 936 00:38:40,840 --> 00:38:41,600 SOMETHING VERY INTERESTING 937 00:38:41,600 --> 00:38:47,000 EMERGED SO SHOWN HERE AT THE 938 00:38:47,000 --> 00:38:48,880 LEFT OF ALL OUR CELLS, ALONG 939 00:38:48,880 --> 00:38:52,360 WITH THOSE FROM PATIENTS WITH 940 00:38:52,360 --> 00:38:55,240 DBA, AND PATIENTS WITH MDS AND 941 00:38:55,240 --> 00:38:58,880 THEY FOLLOW 2 TRAJECTORIES SO 942 00:38:58,880 --> 00:38:59,840 NORMAL CELLS WHO DIFFERENTIATE 943 00:38:59,840 --> 00:39:00,840 NORMALLY WILL GO LIKE THIS AND 944 00:39:00,840 --> 00:39:04,600 COME BACK AND EVENTUALLY DIE 945 00:39:04,600 --> 00:39:06,040 WHEN THEY NUCLEATE AND THEIR 946 00:39:06,040 --> 00:39:09,280 EFFECT, BUT IN DBA AND DEL5 Q 947 00:39:09,280 --> 00:39:11,080 AND IN NORMAL ALSO, THERE'S A 948 00:39:11,080 --> 00:39:12,880 ULTRA TRANSLATIONAL RESEARCH 949 00:39:12,880 --> 00:39:14,600 SCRECT GEE, WHICH DIVERGE RIGHT 950 00:39:14,600 --> 00:39:18,800 AT THE CFU STAGE, AND EARLY 951 00:39:18,800 --> 00:39:20,120 STAGE AND I'LL SHOW YOU THIS IN 952 00:39:20,120 --> 00:39:21,800 THE NEXT SLIDE. 953 00:39:21,800 --> 00:39:24,800 CELLS OF ALL TYPES CONTROL DBA 954 00:39:24,800 --> 00:39:26,480 AND MDS BUT IN DIFFERENT 955 00:39:26,480 --> 00:39:28,640 FREQUENCIES IT'S HARD TO SEE THE 956 00:39:28,640 --> 00:39:30,400 YELLOW AND ORANGE BUT IN 957 00:39:30,400 --> 00:39:31,120 DIFFERENT FREQUENCIES FOLLOW 958 00:39:31,120 --> 00:39:38,240 THIS PATHWAY AS WELL AS THIS 959 00:39:38,240 --> 00:39:38,600 PATHWAY. 960 00:39:38,600 --> 00:39:42,800 AT WHAT EXACTLY HAPPENED TO THE 961 00:39:42,800 --> 00:39:44,920 SINGLE STAGE AND WHAT 962 00:39:44,920 --> 00:39:45,560 CHARACTERISTICS RIGHT AT THIS 963 00:39:45,560 --> 00:39:50,760 BRANCH OF THE CELLS THAT FOLLOW 964 00:39:50,760 --> 00:39:57,600 TRAJECTORY A AND THESE ARE 965 00:39:57,600 --> 00:39:58,000 PERSISTENT TO DIE. 966 00:39:58,000 --> 00:40:09,040 ANOTHER THING I WANT TO POINT 967 00:40:09,040 --> 00:40:10,200 OUT--THE AGENT OF IRON TO GET 968 00:40:10,200 --> 00:40:11,360 INTO CELLS AND HEME TO BE MADE 969 00:40:11,360 --> 00:40:14,120 AND SO THERE WAS GUIDANCE FROM 970 00:40:14,120 --> 00:40:16,240 JUST WHERE THIS EXISTED THAT IT 971 00:40:16,240 --> 00:40:17,840 WAS LIKELY THAT THIS DIVISION AS 972 00:40:17,840 --> 00:40:21,080 I WILL SHOW YOU THIS DIVISION IN 973 00:40:21,080 --> 00:40:22,720 BOTH NORMAL AND ABNORMAL CELLS 974 00:40:22,720 --> 00:40:25,400 RELATES TO HOW THEY HANDLE HEME 975 00:40:25,400 --> 00:40:27,600 AT THIS CRITICAL STAGE 976 00:40:27,600 --> 00:40:28,000 DIFFERENTIATION. 977 00:40:28,000 --> 00:40:31,200 SO IF 1 LOOKS AND DOT PLOTS THE 978 00:40:31,200 --> 00:40:31,800 DIFFERENTIALLY EXPRESSED GENES 979 00:40:31,800 --> 00:40:34,240 AT JUST THOSE THAT ARE GOING 980 00:40:34,240 --> 00:40:35,600 TOWARDS TRAJECTORY A VERSUS 981 00:40:35,600 --> 00:40:38,400 THOSE THAT ARE GOING TOWARDS 982 00:40:38,400 --> 00:40:39,600 TRAJECTORY B,OR CLINICAL 983 00:40:39,600 --> 00:40:42,600 GENETICSINALLY IN SOME CELL 984 00:40:42,600 --> 00:40:44,640 CYCLEOXIDATIVE METABOLISM GENES 985 00:40:44,640 --> 00:40:45,840 ARE MOST UPREGULATED AND THOSE 986 00:40:45,840 --> 00:40:48,040 ON THE HEALTHY PATHWAY AND 987 00:40:48,040 --> 00:40:49,600 TRAJECTORY VIA THE SICK PATHWAY, 988 00:40:49,600 --> 00:40:51,200 THERE'S A RAPID DECREASE IN 989 00:40:51,200 --> 00:40:52,240 CELLULAR GENE EXPRESSIONS OF 990 00:40:52,240 --> 00:40:54,400 PURE GENES AS CELLS ARE DYING 991 00:40:54,400 --> 00:40:56,440 THEY'RE EXPRESSED AND THIS IS 992 00:40:56,440 --> 00:40:58,440 SEEN AS MITOCHONDRIAL GENES 993 00:40:58,440 --> 00:40:59,600 BEING EXPRESSED RELATIVE TO THE 994 00:40:59,600 --> 00:41:00,920 NUMBER OF CELLULAR GENES 995 00:41:00,920 --> 00:41:05,960 EXPRESSED AND HIGHER EXPRESSION 996 00:41:05,960 --> 00:41:07,760 OF A NUMBER OF OF GENES THAT ARE 997 00:41:07,760 --> 00:41:09,400 CORRELATED WITH THEM AND IS ALSO 998 00:41:09,400 --> 00:41:12,560 HIGHER EXPRESSION OF GENES THAT 999 00:41:12,560 --> 00:41:14,120 CORRELATE WITH HEME EXCESS SUCH 1000 00:41:14,120 --> 00:41:17,880 AS RIBOSOMAL PROTEINS SUCH AS 1001 00:41:17,880 --> 00:41:19,880 GENES AND GLOBEIN GENES ARE THE 1002 00:41:19,880 --> 00:41:21,880 MOST HIGHLY EXPRESSED GENES AS 1003 00:41:21,880 --> 00:41:23,560 THE BEGINNING OF THIS PATHWAY. 1004 00:41:23,560 --> 00:41:24,720 SO AS A THIRD SUMMARY BY 1005 00:41:24,720 --> 00:41:26,600 STUDYING THE FATE OF SINGLE 1006 00:41:26,600 --> 00:41:30,120 CELLS, FROM DBA AND DEL5 Q MDS 1007 00:41:30,120 --> 00:41:32,120 PATIENTS AND CONTROLS WE CAN 1008 00:41:32,120 --> 00:41:34,400 FIND A HEALTHY AND 1009 00:41:34,400 --> 00:41:35,800 UNHEALTHYICIDE O TIME 1010 00:41:35,800 --> 00:41:36,080 TRAJECTORY. 1011 00:41:36,080 --> 00:41:40,200 THESE DIVERGE RIGHT AFTER THE 1012 00:41:40,200 --> 00:41:41,760 EPIREGULATION AND HEME 1013 00:41:41,760 --> 00:41:42,000 SYNTHESIS. 1014 00:41:42,000 --> 00:41:42,800 ALTHOUGH DIFFERENTIATION STOPS 1015 00:41:42,800 --> 00:41:44,440 AND CELL DEATH OCCURS MUCH LATER 1016 00:41:44,440 --> 00:41:46,400 AND THAT'S ACTUALLY IMPORTANT 1017 00:41:46,400 --> 00:41:47,680 BECAUSE THE SUGGESTS A WINDOW OF 1018 00:41:47,680 --> 00:41:48,840 TIME BETWEEN WHEN CELLS ARE 1019 00:41:48,840 --> 00:41:51,040 BEGINNING TO MAKE A DECISION 1020 00:41:51,040 --> 00:41:53,960 ABOUT THEIR FATE AND WHEN THAT'S 1021 00:41:53,960 --> 00:41:56,800 ACIVATED AT THE PROTEIN LEVEL 1022 00:41:56,800 --> 00:41:58,560 AND CELL LEVEL AND SO 1023 00:41:58,560 --> 00:42:03,000 POTENTIALLY A TIME THAT 1 COULD 1024 00:42:03,000 --> 00:42:03,840 INTERVENE THERAPEUTICALLY WITH 1025 00:42:03,840 --> 00:42:04,080 SUCCESS. 1026 00:42:04,080 --> 00:42:06,160 THE CELLS THAT ARE DESTINED TO 1027 00:42:06,160 --> 00:42:08,960 DIE ARE IN HEME REGULATED GENES 1028 00:42:08,960 --> 00:42:11,040 AND IN CONTRAST SURVIVING CELLS 1029 00:42:11,040 --> 00:42:13,400 DOWN REGULATE HEEM SYNTHESIS 1030 00:42:13,400 --> 00:42:14,600 WHILE UPREGULATING 1031 00:42:14,600 --> 00:42:17,000 PHOSPHORYLATION AND OTHER GENES 1032 00:42:17,000 --> 00:42:18,000 PROCESS, PERHAPS SUGGESTING 1033 00:42:18,000 --> 00:42:20,120 TARGETS FOR THERAPY THAT COULD 1034 00:42:20,120 --> 00:42:27,080 MODULATE THIS CHOICE AND THEN 1035 00:42:27,080 --> 00:42:28,800 QUITE UNEXPECTEDLY BETWEEN 1036 00:42:28,800 --> 00:42:32,120 19-31% OF CONTROL CELLS FOLLOW 1037 00:42:32,120 --> 00:42:32,800 THE UNHEALTHY TRAJECTORY, 1038 00:42:32,800 --> 00:42:37,520 IMPLYING THAT HEME IS ALSO A 1039 00:42:37,520 --> 00:42:42,000 RHEOSTAT THAT REGS REGS--REGULATES 1040 00:42:42,000 --> 00:42:42,560 RED CELL PRODUCTION. 1041 00:42:42,560 --> 00:42:43,200 AND WHEN YOU THINK ABOUT IT, 1042 00:42:43,200 --> 00:42:44,960 THEY HAVE TO BE READY TO MAKE SO 1043 00:42:44,960 --> 00:42:47,040 MUCH MORE IN A SITUATION OF 1044 00:42:47,040 --> 00:42:52,920 ACUTE ANEMIA AND THEN CELLS ARE 1045 00:42:52,920 --> 00:42:54,600 STAGED AT THE CFUE LEVEL AND 1046 00:42:54,600 --> 00:42:57,960 THEN CAN MATURE LATER SO UNDER 1047 00:42:57,960 --> 00:42:58,680 NORMAL CONDITIONS THEY ARE 1048 00:42:58,680 --> 00:43:00,440 STAGED AND THEN WE PROTECT THEM 1049 00:43:00,440 --> 00:43:03,280 AND HAVE THEM MATURE AND EXPAND 1050 00:43:03,280 --> 00:43:04,520 VERY QUICKLY BECAUSE THEY'RE ALL 1051 00:43:04,520 --> 00:43:06,360 LEARNING PART WAY ALONG THE 1052 00:43:06,360 --> 00:43:08,040 DIFFERENTIATION SEQUENCE AND SO 1053 00:43:08,040 --> 00:43:08,720 PHYSIOLOGICALLY IN ORDER TO 1054 00:43:08,720 --> 00:43:10,520 ALLOW US TO HAVE QUICK RESPONSE 1055 00:43:10,520 --> 00:43:12,400 TO ANEMIA, WE SACRIFICE A LOT OF 1056 00:43:12,400 --> 00:43:13,920 CELLS AT THE STAGE 1057 00:43:13,920 --> 00:43:15,680 DIFFERENTIATION TO HAVE THEM IN 1058 00:43:15,680 --> 00:43:18,120 RESERVE FOR THESE IMPORTANT 1059 00:43:18,120 --> 00:43:20,760 CIRCUMSTANCES THAT 1 NEEDS TO 1060 00:43:20,760 --> 00:43:21,200 RESPOND TO. 1061 00:43:21,200 --> 00:43:24,600 AND I GUESS A SECOND MESSAGE OF 1062 00:43:24,600 --> 00:43:26,960 THAT IS THAT MANY OF US OVER OUR 1063 00:43:26,960 --> 00:43:28,560 LIFETIMES HAVE THOUGHT ABOUT 1064 00:43:28,560 --> 00:43:30,400 DIFFERENTIATION OR SPECIFIC RED 1065 00:43:30,400 --> 00:43:31,560 CELL DIFFERENTIATION, AS THIS 1066 00:43:31,560 --> 00:43:33,000 STAGE AND THAT STAGE, AND THIS 1067 00:43:33,000 --> 00:43:37,400 STAGE IS THOSE CELLS STOP AND AS 1068 00:43:37,400 --> 00:43:41,240 A POPULATION ARE ACTUALLY 1069 00:43:41,240 --> 00:43:42,360 DIFFERENTIATABLE IN A DISCREET 1070 00:43:42,360 --> 00:43:44,880 WAY BUT ACTUALLY IT'S A VERY 1071 00:43:44,880 --> 00:43:48,280 DINETTIC PROCESS, A VERY FAST 1072 00:43:48,280 --> 00:43:49,120 DINETTIC PROCESS AND INDIVIDUAL 1073 00:43:49,120 --> 00:43:50,920 CELLS HAVE TO MAKE QUICK 1074 00:43:50,920 --> 00:43:53,360 DECISIONS AND THE PROCESS 1075 00:43:53,360 --> 00:43:54,280 OVERALL PHYSIOLOGICALLY ALLOWS 1076 00:43:54,280 --> 00:43:56,200 FOR LOTS OF CELLS TO DIE AND BE 1077 00:43:56,200 --> 00:43:58,600 EFFECTIVE AND TO ASSURE THAT RED 1078 00:43:58,600 --> 00:43:59,760 CELL DIFFERENTIATION IS NOT ONLY 1079 00:43:59,760 --> 00:44:03,600 EFFECTIVE BUT CAN EXPAND 5-10 1080 00:44:03,600 --> 00:44:04,200 FOLD. 1081 00:44:04,200 --> 00:44:09,600 AND THEN LASTLY THE DEL5 Q MDS 1082 00:44:09,600 --> 00:44:14,520 UNLIKE DBA AS NORMAL MUTATION 1083 00:44:14,520 --> 00:44:17,360 MEANING AND PRESENTATION FOR 1084 00:44:17,360 --> 00:44:19,840 MDS, ONLY ABOUT 50-75% OF CELLS 1085 00:44:19,840 --> 00:44:22,000 CONTAIN THE LACK OF CHROMOSOME 1086 00:44:22,000 --> 00:44:22,320 52. 1087 00:44:22,320 --> 00:44:25,360 AND YET THE NORMAL CELLS DON'T 1088 00:44:25,360 --> 00:44:27,400 DIFFERENTIATE AT ALL, WELL, AND 1089 00:44:27,400 --> 00:44:28,760 SO BECAUSE PRESENTATIONS WITH 1090 00:44:28,760 --> 00:44:30,400 ANEMIA, SO TRACKING THEIR 1091 00:44:30,400 --> 00:44:32,480 TRAJECTORY SHOULD PROVIDE NEW 1092 00:44:32,480 --> 00:44:34,640 INSIGHTS INTO WHY THEY CAN'T 1093 00:44:34,640 --> 00:44:35,600 COMPENSATE, WHY THEY CAN'T 1094 00:44:35,600 --> 00:44:37,160 EXPAND AND WHY THEY CAPTAIN 1095 00:44:37,160 --> 00:44:38,400 PREVENT ANEMIA AND THIS IS 1096 00:44:38,400 --> 00:44:39,440 SOMETHING ELSE THAT WE'RE 1097 00:44:39,440 --> 00:44:41,080 ACTUALLY ABLE TO DO BECAUSE 1 1098 00:44:41,080 --> 00:44:43,160 CAN LOOK AT THE AMOUNT OF 1099 00:44:43,160 --> 00:44:46,120 EXPRESSION OF THOSE GENES THAT 1100 00:44:46,120 --> 00:44:50,080 ARE ON CHROMOSOME 5 Q AND 1101 00:44:50,080 --> 00:44:51,120 SEPARATE CELLS IN THE 1102 00:44:51,120 --> 00:44:51,920 [INDISCERNIBLE] OF THE PATIENT 1103 00:44:51,920 --> 00:44:54,600 THAT COMES TO THE MDS, INTO 1104 00:44:54,600 --> 00:44:56,000 THOSE CELLS THAT HAVE PRESERVED 1105 00:44:56,000 --> 00:44:58,640 5 Q AND THOSE THAT DON'T. 1106 00:44:58,640 --> 00:45:01,000 AND INTERESTINGLY, THE 5 Q 1107 00:45:01,000 --> 00:45:02,480 PRESERVED CELLS, THE 5 + CELLS 1108 00:45:02,480 --> 00:45:06,560 FROM AN MDS CELLS WITH DEL5 Q 1109 00:45:06,560 --> 00:45:10,600 MDS, ON A HEALTHY TRAJECTORY, 1110 00:45:10,600 --> 00:45:15,560 THEY ARE PRETTY NORMAL AND THOSE 1111 00:45:15,560 --> 00:45:16,840 TRANSCRIPTOMES ARE DIFFERENT 1112 00:45:16,840 --> 00:45:19,000 FROM THE STUDIES THAT ARE DONE 1113 00:45:19,000 --> 00:45:19,760 SIMULTANEOUSLY AND THIS 1114 00:45:19,760 --> 00:45:21,080 IBDICATES THAT THE M, DID S 1115 00:45:21,080 --> 00:45:26,600 PATIENT CELLS WITH THE DEL5 1116 00:45:26,600 --> 00:45:28,200 DELETION DIRECTLY OR INDIRECTLY, 1117 00:45:28,200 --> 00:45:29,400 PERHAPS BECAUSE OF THE 1118 00:45:29,400 --> 00:45:30,200 ENVIRONMENT AROUND THEM, BE 1119 00:45:30,200 --> 00:45:34,600 IMPAIRED AND SO THE EFFECTIVE 1120 00:45:34,600 --> 00:45:35,840 DIFFERENTIATION OF NEIGHBORING 1121 00:45:35,840 --> 00:45:38,080 GENETICALLY NORMAL CELLS ARE 1122 00:45:38,080 --> 00:45:39,640 IMPAIRED IN AN ENVIRONMENT WHEN 1123 00:45:39,640 --> 00:45:43,200 DEL5 Q IS PRESENT AND 1124 00:45:43,200 --> 00:45:43,640 INTERESTINGLYOXIDATIVE 1125 00:45:43,640 --> 00:45:44,640 PHOSPHORYLATION RNA SPICE AND 1126 00:45:44,640 --> 00:45:47,080 CELL CYCLE CHECK POINTS ARE ON A 1127 00:45:47,080 --> 00:45:48,880 RIBOSOME PRODUCTION, A HEF1 1128 00:45:48,880 --> 00:45:49,720 SIGNALING PATHWAYS ARE ALTERED 1129 00:45:49,720 --> 00:45:52,600 AND THESE ARE ALL SIMILAR 1130 00:45:52,600 --> 00:45:53,920 FINDINGS TO PATIENTS 5 Q MINUS 1131 00:45:53,920 --> 00:45:55,840 CELLS AND TO THE CELLS FROM DBA 1132 00:45:55,840 --> 00:45:59,960 PATIENTS AND TO THE CELLS FROM 1133 00:45:59,960 --> 00:46:01,600 THE MICE WITH DBA AND CELLS FROM 1134 00:46:01,600 --> 00:46:05,520 THE MICE WITH LACKING FLVCR, 1135 00:46:05,520 --> 00:46:07,760 SUGGESTING THAT I TOO HAVE HUMAN 1136 00:46:07,760 --> 00:46:09,160 DAMAGE EVEN THOAR THEY'RE 1137 00:46:09,160 --> 00:46:09,880 GENETICALLY NORMAL. 1138 00:46:09,880 --> 00:46:12,840 SO THAT'S ACTUALLY LED TO NEW 1139 00:46:12,840 --> 00:46:14,040 QUESTIONS THAT WE'RE ADDRESSING 1140 00:46:14,040 --> 00:46:15,440 IN THE LABORATORY TO TRY TO 1141 00:46:15,440 --> 00:46:17,480 FIGURE OUT WHAT THAT EFFECT 1142 00:46:17,480 --> 00:46:21,760 MIGHT BE AND HOW COULD IT RELATE 1143 00:46:21,760 --> 00:46:22,160 TO HEME. 1144 00:46:22,160 --> 00:46:24,000 SO KIND OF AS A LAST SUMMARY, I 1145 00:46:24,000 --> 00:46:27,320 WANT TO GET BACK TO CLINICAL 1146 00:46:27,320 --> 00:46:27,520 WORK. 1147 00:46:27,520 --> 00:46:29,160 AND I KNOW THAT I TALKED EARLIER 1148 00:46:29,160 --> 00:46:31,800 TO A NUMBER OF FOLK THAT HAVE 1149 00:46:31,800 --> 00:46:33,760 HAD THEIR SECOND YOOER 1150 00:46:33,760 --> 00:46:34,600 EMATTOLOGY IN SECOND YEAR 1151 00:46:34,600 --> 00:46:36,920 MEDICAL SCHOOL AND HAVE BASIC 1152 00:46:36,920 --> 00:46:39,400 HEMATOLOGY TRAINING AND KNOW 1153 00:46:39,400 --> 00:46:44,000 THAT A NORMAL RED CELL IS IN A 1154 00:46:44,000 --> 00:46:45,600 NUCLEUS SACK, IT DOESN'T CONTAIN 1155 00:46:45,600 --> 00:46:48,880 A LOT, IT'S 95% OF ITS PROTEIN 1156 00:46:48,880 --> 00:46:50,640 CONTENT AND RED CELL IS NORMAL 1157 00:46:50,640 --> 00:46:53,760 ACIDIC AND WHICH IS SIZE AND 1158 00:46:53,760 --> 00:46:57,040 CELL VOLUME IS BETWEEN 80 AND A 1159 00:46:57,040 --> 00:46:57,520 HUNDRED. 1160 00:46:57,520 --> 00:47:00,520 AND OF COURSE AS MCHCSO THE 1161 00:47:00,520 --> 00:47:01,800 CONCENTRATION IS NORMAL BECAUSE 1162 00:47:01,800 --> 00:47:04,240 THAT'S WHAT'S REQUIRED FOR THE 1163 00:47:04,240 --> 00:47:06,320 EFFECTIVE IRON DELIVERY AND FOR 1164 00:47:06,320 --> 00:47:06,800 CO2 EXCHANGE. 1165 00:47:06,800 --> 00:47:09,720 AND WE ALL KNOW TO LOOK AT 1166 00:47:09,720 --> 00:47:11,600 MICROSITTIC RED CELLS AS SMALLER 1167 00:47:11,600 --> 00:47:14,440 THAT LYMPHOCYTES SHOWN HERE AND 1168 00:47:14,440 --> 00:47:16,560 HYPOCHROMIC MEANING HAVING LESS 1169 00:47:16,560 --> 00:47:18,640 HEMOGLOBIN, AND WE CAN USE THAT 1170 00:47:18,640 --> 00:47:22,200 AS A CRITERIA FOR LOOKING AT RNA 1171 00:47:22,200 --> 00:47:25,200 SUFFICIENCY OR FOR REALLY FAILED 1172 00:47:25,200 --> 00:47:27,440 HEME PRODUCTION SUCH AS HEME 1173 00:47:27,440 --> 00:47:32,200 SUFFICIENCY OR FAILED PRODUCTION 1174 00:47:32,200 --> 00:47:33,800 SUCH AS THALACEMIA, AND IT MAKES 1175 00:47:33,800 --> 00:47:34,680 ITSELF SMALLER BECAUSE IT'S 1176 00:47:34,680 --> 00:47:45,280 REALLY IMPORTANT TO TRY TO 1177 00:47:45,280 --> 00:47:47,400 PRESERVE AND FOR THE RED CELL 1178 00:47:47,400 --> 00:47:48,880 EXCHANGE FOR CO2. 1179 00:47:48,880 --> 00:47:51,800 AND CELLS TRY WELL WITH WELL 1180 00:47:51,800 --> 00:47:52,600 UNDERSTOOD MOLECULAR MECHANISMS 1181 00:47:52,600 --> 00:47:55,480 TO CONTROL A NORMAL MCHC BAY 1182 00:47:55,480 --> 00:47:57,560 MAKING NORMAL CELLS IN A SMALLER 1183 00:47:57,560 --> 00:47:58,320 SITUATION OF IRON DEFICIENCY. 1184 00:47:58,320 --> 00:47:59,920 SO I WANT TO LEAVE YOU WITH THE 1185 00:47:59,920 --> 00:48:01,200 THOUGHT THAT THE ANEMIA ARE THAT 1186 00:48:01,200 --> 00:48:03,000 ARE EFFECTIVE THAT I TALKED 1187 00:48:03,000 --> 00:48:04,960 ABOUT ARE MACROSITTIC, SO THE 1188 00:48:04,960 --> 00:48:09,080 MCV IS LARGER THAN 100. 1189 00:48:09,080 --> 00:48:12,400 AND SO IF THERE'S AN ABUNDANCE 1190 00:48:12,400 --> 00:48:14,200 OF EXCESS HEME ARE CELLS MADE 1191 00:48:14,200 --> 00:48:15,000 LARGER IN COMPENSATION IN ORDER 1192 00:48:15,000 --> 00:48:20,680 TO TRY TO MAINTAIN A CELL SYSTEM 1193 00:48:20,680 --> 00:48:22,720 THAT'S OPTIMUM FOR DELIVERY AND 1194 00:48:22,720 --> 00:48:23,520 SO THE CELLULAR AND EXTRA 1195 00:48:23,520 --> 00:48:26,360 CELLULAR TO MAKE SURE THIS 1196 00:48:26,360 --> 00:48:28,120 HAPPENS, SO THAT ON A LARGER 1197 00:48:28,120 --> 00:48:30,440 SCALE, NOT THAT SCALE OF HEME 1198 00:48:30,440 --> 00:48:31,280 DIFFERENTIATION AND CELLS, BUT 1199 00:48:31,280 --> 00:48:33,560 THE CELLS IN THE ORGANISMS OF A 1200 00:48:33,560 --> 00:48:35,080 PERSON OR OF A MOUSE OR OF A CAT 1201 00:48:35,080 --> 00:48:37,840 OR WHATEVER YOU'RE TALKING 1202 00:48:37,840 --> 00:48:42,600 ABOUT, THAT THERE'S OPTIMAL OF 1203 00:48:42,600 --> 00:48:48,560 AND OPTIMAL EXCHANGE OF CO2. 1204 00:48:48,560 --> 00:48:51,640 THE HEME ARE IN BALANCE AS THESE 1205 00:48:51,640 --> 00:48:52,040 CELLS WERE MADE. 1206 00:48:52,040 --> 00:48:53,480 SO WITH THAT I WOULD LIKE TO 1207 00:48:53,480 --> 00:48:55,400 THANK THE BULK OF MY LAB, THIS 1208 00:48:55,400 --> 00:49:06,080 IS ACTUALLY AN OLD PICTURE, 1209 00:49:06,080 --> 00:49:07,160 GENERATED OVER MANY YEARS, FOR 1210 00:49:07,160 --> 00:49:09,800 THOSE OF YOU THAT DON'T KNOW THE 1211 00:49:09,800 --> 00:49:11,960 WEST COAST IN SEATTLE WE'RE HALF 1212 00:49:11,960 --> 00:49:13,480 DAY TRIP FROM 3 DIFFERENT 1213 00:49:13,480 --> 00:49:15,440 NATIONAL PARKS, AND THIS IS 1 OF 1214 00:49:15,440 --> 00:49:15,680 THEM. 1215 00:49:15,680 --> 00:49:21,600 AND I LIKE TO PARTICULARLY THANK 1216 00:49:21,600 --> 00:49:24,440 ZATO YANG WHO DID THE FIRST WORK 1217 00:49:24,440 --> 00:49:27,440 THAT I SHOWED YOU ON THE SCIENCE 1218 00:49:27,440 --> 00:49:31,960 TRANSLATIONAL MEDICINE PAPER TO 1219 00:49:31,960 --> 00:49:32,760 RAYMOND DOTY, AND IS RESPONSIBLE 1220 00:49:32,760 --> 00:49:37,320 FOR ALL THE MOUSE AND MODEL 1221 00:49:37,320 --> 00:49:38,040 CITIZEN LEAKULAR WORK. 1222 00:49:38,040 --> 00:49:44,040 LOOKING FORWARD TO COLLABORATORS 1223 00:49:44,040 --> 00:49:46,800 CHRIS LOSTED, AND INFORMATICS, 1224 00:49:46,800 --> 00:49:48,920 PERSON THAT IS BEEN A SUPER 1225 00:49:48,920 --> 00:49:54,600 COLLABORATORS IN TERMS OF SINGLE 1226 00:49:54,600 --> 00:49:55,880 CELL ANALYSIS AND 1227 00:49:55,880 --> 00:49:56,600 [INDISCERNIBLE] WHO IS WORKING 1228 00:49:56,600 --> 00:49:58,800 THE ISSUE OF WHY ARE NORMAL 1229 00:49:58,800 --> 00:50:01,720 CELLS ABNORMAL IN THE 1230 00:50:01,720 --> 00:50:03,360 ENVIRONMENT OF DEL5 Q MDS, AND 1231 00:50:03,360 --> 00:50:07,200 OF COURSE OUR MANY SUPPORTS FROM 1232 00:50:07,200 --> 00:50:09,480 R01s FROM THE NIH FOR OVER 30 1233 00:50:09,480 --> 00:50:11,520 YEARS AND FROM OTHER FOUNDATIONS 1234 00:50:11,520 --> 00:50:13,000 AND THANK YOU VERY MUCH FOR YOUR 1235 00:50:13,000 --> 00:50:13,280 ATTENTION. 1236 00:50:13,280 --> 00:50:18,400 I'M HAPPY TO ANSWER QUESTIONS. 1237 00:50:18,400 --> 00:50:21,080 >> THANKS, JAN. 1238 00:50:21,080 --> 00:50:21,320 SORRY. 1239 00:50:21,320 --> 00:50:23,560 AS ALWAYS, VERY CLEAR AND 1240 00:50:23,560 --> 00:50:24,800 STIMULATING, SO THERE IS A 1241 00:50:24,800 --> 00:50:25,560 QUESTION FROM [INDISCERNIBLE] 1242 00:50:25,560 --> 00:50:28,000 WHO IS A YOUNG INVESTIGATOR AT 1243 00:50:28,000 --> 00:50:28,920 NHLBI AND CARDIOLOGYST BUT WE 1244 00:50:28,920 --> 00:50:32,000 STILL TALK TO HIM AND HE WANTS 1245 00:50:32,000 --> 00:50:33,320 TO KNOW,--THANKS FOR SHARING 1246 00:50:33,320 --> 00:50:35,200 THESE INTERESTING DATA ARE THERE 1247 00:50:35,200 --> 00:50:36,600 INSTANCES OF SOMATIC MUTATIONS 1248 00:50:36,600 --> 00:50:39,160 ANOTHER THAN 5 Q MINUS THAT 1249 00:50:39,160 --> 00:50:41,440 AFFECT THE FUNCTIONS 1250 00:50:41,440 --> 00:50:42,240 SPECIFICALLY IN ERYTHROPOIESIS 1251 00:50:42,240 --> 00:50:44,280 AND HAVE CLINICAL IMPLICATIONS? 1252 00:50:44,280 --> 00:50:47,240 >> YEAH, THE ANSWER IS PROBABLY 1253 00:50:47,240 --> 00:50:49,480 YES, BUT THE MORE IMPORTANT 1254 00:50:49,480 --> 00:50:51,520 ANSWER TO ME IN TERMS OF 1255 00:50:51,520 --> 00:50:52,920 THINKING OF ERYTHROPOIESIS IS 1256 00:50:52,920 --> 00:50:55,760 THAT ANYTHING THAT IMPAIRS 1257 00:50:55,760 --> 00:50:56,720 PROTEIN BEING MADE WHETHER IT'S 1258 00:50:56,720 --> 00:50:58,760 THE RIEB SO STUDIES OF MULTIPLE 1259 00:50:58,760 --> 00:51:00,040 ENDOCRINE THAT'S IMPAIRED OR 1260 00:51:00,040 --> 00:51:05,000 DIRECT IMPAIRMENT OF PROTEIN 1261 00:51:05,000 --> 00:51:05,720 TRANSCRIPTION AND TRANSLATION 1262 00:51:05,720 --> 00:51:07,800 FOR SOME OTHER MECHANISM WILL 1263 00:51:07,800 --> 00:51:10,000 MAKE FOR ADEQUATE HEME BECAUSE 1264 00:51:10,000 --> 00:51:11,200 IT REQUIRES SUCH LITTLE PROTEIN 1265 00:51:11,200 --> 00:51:13,760 TO GET THERE BECAUSE IT'S AN 1266 00:51:13,760 --> 00:51:14,800 ENZYME ATDIC PROCESS, AND YOU 1267 00:51:14,800 --> 00:51:16,800 HAVE PLENTY OF HEAT, BUT IS 1268 00:51:16,800 --> 00:51:18,320 GOING TO IMPAIR THE AMOUNT OF 1269 00:51:18,320 --> 00:51:21,800 DPLOABIN THAT YOU NEED, BECAUSE 1270 00:51:21,800 --> 00:51:23,640 YOU NEED OODLES OF GLOBEIN TO 1271 00:51:23,640 --> 00:51:25,560 BALANCE THE HEME BECAUSE IT'S A 1272 00:51:25,560 --> 00:51:27,200 PROTEIN SO YOU HAVE TO HAVE 1273 00:51:27,200 --> 00:51:28,000 EFFICIENT PROTEIN PRODUCTION, SO 1274 00:51:28,000 --> 00:51:31,160 I THINK OF IT MORE GENERICALLY 1275 00:51:31,160 --> 00:51:32,560 THAT MANY THINGS WILL DO THIS 1276 00:51:32,560 --> 00:51:38,120 AND THIS LIKELY IS A REASON 1277 00:51:38,120 --> 00:51:39,800 CONTRIBUTING TO ANEMIA OR 1278 00:51:39,800 --> 00:51:43,600 COMPONENT OF ANEMIA IN LOTS OF 1279 00:51:43,600 --> 00:51:44,720 DIFFERENT SETTINGS. 1280 00:51:44,720 --> 00:51:46,800 THERE IS MORE OBVIOUS 1S SUCH AS 1281 00:51:46,800 --> 00:51:50,240 LOOKING AT THE ANEMIA IN 1282 00:51:50,240 --> 00:51:51,600 THALCEMIA WHICH IS PROBABLY 1283 00:51:51,600 --> 00:51:53,560 DOUBLE, IT HAS INEFFECTIVE 1284 00:51:53,560 --> 00:51:54,880 ERYTHROPOIESIS BECAUSE OF THE 1285 00:51:54,880 --> 00:51:58,040 FACT YOU DON'T MAYBE GLOBEIN 1286 00:51:58,040 --> 00:51:59,720 BECAUSE OR YOU DON'T MAKE IT 1287 00:51:59,720 --> 00:52:02,360 WELL BECAUSE OF THE DISCONNECT 1288 00:52:02,360 --> 00:52:03,960 WITH THE GLOBEIN CHAINS SO 1289 00:52:03,960 --> 00:52:05,160 THERE'S EXCESS OF GLOBEIN LIKE 1290 00:52:05,160 --> 00:52:07,960 WHAT I TALKED ABOUT AND THEN 1291 00:52:07,960 --> 00:52:09,880 THERE'S SECOND SUCCESS LATER 1292 00:52:09,880 --> 00:52:11,520 BECAUSE THEY'RE MISMATCHED AND 1293 00:52:11,520 --> 00:52:12,400 INEFFECTIVENESS AT THAT STAGE. 1294 00:52:12,400 --> 00:52:15,240 SO I THINK OF ANEMIA AS BEING SO 1295 00:52:15,240 --> 00:52:17,960 MANY THINGS YOU HAVE TO 1296 00:52:17,960 --> 00:52:21,040 ACCOMPLISH TO HAVE A CELL COME 1297 00:52:21,040 --> 00:52:22,880 UP ABRIEWPTLY AND SUBTLE CHANGE 1298 00:52:22,880 --> 00:52:25,480 CANS MAKE FOR SUBTLE ANEMIAS AND 1299 00:52:25,480 --> 00:52:27,080 DIRECT CHANGES PARTICULARLY IN 1300 00:52:27,080 --> 00:52:28,200 TRANSCRIPTION AND TRANSLATION, 1301 00:52:28,200 --> 00:52:30,720 AT EVERY LEVEL AND RIBOSOMAL 1302 00:52:30,720 --> 00:52:32,480 ABNORMALITY CAN MAKE FOR THIS 1303 00:52:32,480 --> 00:52:34,600 DISCONNECT THAT I SPOKE ABOUT. 1304 00:52:34,600 --> 00:52:34,960 >> GREAT. 1305 00:52:34,960 --> 00:52:37,320 THANKS, JAN, NOW THERE'S A 1306 00:52:37,320 --> 00:52:38,080 QUESTION FROM CAROLYN 1307 00:52:38,080 --> 00:52:40,000 [INDISCERNIBLE] WHO I'M SURE YOU 1308 00:52:40,000 --> 00:52:47,160 ALSO KNOW, IS HEME REGULATION 1309 00:52:47,160 --> 00:52:50,320 ALL 32 BLOCK 1--IMPROVE THE 1310 00:52:50,320 --> 00:52:51,480 ANEMIA IN DBA? 1311 00:52:51,480 --> 00:52:53,880 >> SO THE ANSWER ISIO AND YES. 1312 00:52:53,880 --> 00:52:55,600 IN THAT ORDER. 1313 00:52:55,600 --> 00:52:57,280 IT'S NOT EXCLUSIVELY THROUGH 1314 00:52:57,280 --> 00:52:58,800 BACHR 1 AND THERE'S INDEPENDENT 1315 00:52:58,800 --> 00:53:01,400 FAGHT WAYS AND LOOKEDDA THE THEM 1316 00:53:01,400 --> 00:53:03,800 INDEPENDENTLY AND BOTH ARE 1317 00:53:03,800 --> 00:53:04,320 PROMOTIONAL IMPORTANT. 1318 00:53:04,320 --> 00:53:08,320 WHEN I SHOWED YOU THE GATAC1 1319 00:53:08,320 --> 00:53:12,880 TARGETS, I USED 1 THAT HAD BEEN 1320 00:53:12,880 --> 00:53:14,520 VALIDATED AS TARGETS IN MULTIPLE 1321 00:53:14,520 --> 00:53:16,640 SO WE KNEW FOR SURE THEY WERE IN 1322 00:53:16,640 --> 00:53:18,040 MULTIPLE TARGETS THEY CAME FROM 1323 00:53:18,040 --> 00:53:19,560 THE [INDISCERNIBLE] DATA SETS 1324 00:53:19,560 --> 00:53:22,360 AND SO THEY'VE BEEN PRETTY CLEAN 1325 00:53:22,360 --> 00:53:27,600 AT LOOKING AT BOTH BACH1 AND 1326 00:53:27,600 --> 00:53:28,360 BACH1 INDEPENDENT TARGETS. 1327 00:53:28,360 --> 00:53:30,800 IN TERMS OF WHETHER REDUCING 1328 00:53:30,800 --> 00:53:33,200 IRON CAN HELP, YES, WE'VE DONE 1329 00:53:33,200 --> 00:53:37,600 THAT IN OUR ANIMAL MODELS AND 1330 00:53:37,600 --> 00:53:40,400 REDUCING WAYS OF IRON YOU CAN 1331 00:53:40,400 --> 00:53:43,040 REDUCE HEMEAT O POETICESIS 1332 00:53:43,040 --> 00:53:45,200 THROUGH THE DBA MODEL AND YOU 1333 00:53:45,200 --> 00:53:48,360 CAN IMPROVE HEMEAT O POETICESIS 1334 00:53:48,360 --> 00:53:49,320 OF THE FLVCR SUFFICIENT MOUSE 1335 00:53:49,320 --> 00:53:51,200 AND THE WAY YOU CAN DO THAT 1336 00:53:51,200 --> 00:53:52,600 LIMITING IRON IN THE DIET AND 1337 00:53:52,600 --> 00:53:58,480 BREED THE MICE TO A TRANSFERRED 1338 00:53:58,480 --> 00:53:59,520 RECEPTOR HAPLOID RECEPTOR MOUSE, 1339 00:53:59,520 --> 00:54:01,400 SO LESS GETS IN THE MODELS, CAN 1340 00:54:01,400 --> 00:54:03,760 YOU CAN IN ANIMALS, THAT WE HAVE 1341 00:54:03,760 --> 00:54:09,680 AS WELL AS CELLS TREAT THEM WITH 1342 00:54:09,680 --> 00:54:12,480 SUSASLE TONE, SO IT'S NOT A 1343 00:54:12,480 --> 00:54:14,280 USEFUL THING FOR THINKING ABOUT 1344 00:54:14,280 --> 00:54:16,000 HUMAN TRIAL, BUT I THINK THE 1345 00:54:16,000 --> 00:54:18,360 MOST INTERESTING EXPERIMENT IN 1346 00:54:18,360 --> 00:54:22,120 HUMANS IS ACTUALLY NOT WITH PER 1347 00:54:22,120 --> 00:54:23,440 SE WITH DBA, IN TERMS OF WHAT 1348 00:54:23,440 --> 00:54:26,760 THE ROLE OF HIRING MIGHT BE IN 1349 00:54:26,760 --> 00:54:31,000 LIMITING HEME SYNTHESIS, BUT 1350 00:54:31,000 --> 00:54:33,200 WITH A CONGENITAL PORFERIA, SO 1351 00:54:33,200 --> 00:54:34,960 YOU HAVE ACCUMULATION OF HEEM 1352 00:54:34,960 --> 00:54:37,800 LONG THE PATHWAY LEADING TO THE 1353 00:54:37,800 --> 00:54:38,760 MANIFESTATION SUPPORT, AND BY 1354 00:54:38,760 --> 00:54:41,360 MAKING A PATIENT WITH CEP IRON 1355 00:54:41,360 --> 00:54:43,680 DEFICIENT OR A MOUSE WITH CEP 1356 00:54:43,680 --> 00:54:45,920 IRON DEFICIENT AND YOU CAN 1357 00:54:45,920 --> 00:54:47,000 AMELIORATE THOSE SYMPTOMS SO WE 1358 00:54:47,000 --> 00:54:48,560 AND OTHER VS ACTUALLY PUBLISHED 1359 00:54:48,560 --> 00:54:50,800 ON THAT AS A THERAPEUTIC 1360 00:54:50,800 --> 00:54:57,640 APPROACH TO THIS UNUSUAL PORFER 1361 00:54:57,640 --> 00:54:58,040 IA. 1362 00:54:58,040 --> 00:55:00,920 WE DO PLAN STUDIES, OUR OWN 1363 00:55:00,920 --> 00:55:02,960 STUDIES WITH IRON ACCUMULATION, 1364 00:55:02,960 --> 00:55:04,840 GOT OFF TO SORT OF A ROCKY START 1365 00:55:04,840 --> 00:55:09,320 BECAUSE OF HAD SOME ISSUES WITH 1366 00:55:09,320 --> 00:55:10,000 COMPANY CONNECTION BUT--I DON'T 1367 00:55:10,000 --> 00:55:12,000 KNOW IF YOU WANT TO COMMENT HAS 1368 00:55:12,000 --> 00:55:15,480 TAKEN THIS UP AS A TRAUMA PEG 1369 00:55:15,480 --> 00:55:17,600 AND CERTAINLY GOING AHEAD, WE DO 1370 00:55:17,600 --> 00:55:19,880 PLAN TO THINK ABOUT KEYALATION, 1371 00:55:19,880 --> 00:55:22,720 AND PARTICULARLY KEYALATION THAT 1372 00:55:22,720 --> 00:55:28,600 COULD BE DONE WITH IRON 1373 00:55:28,600 --> 00:55:31,000 KEYLATERS THAT ARE KEY LATE 1374 00:55:31,000 --> 00:55:34,000 REGULARLY, SO IF 1 USED 1375 00:55:34,000 --> 00:55:40,040 [INDISCERNIBLE] THAT 1 CAN KEY 1376 00:55:40,040 --> 00:55:43,400 LATE ERYTHRIN TO THE CELLS MORE 1377 00:55:43,400 --> 00:55:45,240 EASILY, AND SO RED CELLS SHOULD 1378 00:55:45,240 --> 00:55:48,160 HAVE MORE IRON WITHOUT CAUSING A 1379 00:55:48,160 --> 00:55:49,280 VERY SIGNIFICANT IRON DEFICIENCY 1380 00:55:49,280 --> 00:55:55,320 MORE BROADLY, SO THAT IS A 1381 00:55:55,320 --> 00:55:56,760 THERAPEUTIC STRATEGY. 1382 00:55:56,760 --> 00:56:00,000 YEAH, AND WE DO HAVE A CLINICAL 1383 00:56:00,000 --> 00:56:01,400 TRIAL TRYING TO STRATEGY IN DBA 1384 00:56:01,400 --> 00:56:04,000 WITH A DRUG THAT'S VERY POTENT, 1385 00:56:04,000 --> 00:56:05,800 BUT UNFORTUNATELY ALSO DRAWS 1386 00:56:05,800 --> 00:56:07,400 PLATELET COUNTS UP SO WE'VE 1387 00:56:07,400 --> 00:56:09,280 ENDED UP NOT BEING ABLE TO GET 1388 00:56:09,280 --> 00:56:10,760 TO THE PROPER EFFECTIVE DOSES IN 1389 00:56:10,760 --> 00:56:13,400 THOSE PATIENTS BUT AN UNUSUAL 1390 00:56:13,400 --> 00:56:15,960 PATIENT WHO'S IN MOSAIC, WITH 1391 00:56:15,960 --> 00:56:17,360 THE CELL IS ACTIVE--OOPS SORRY 1392 00:56:17,360 --> 00:56:19,360 MY DOG IS VERY EXCITED ABOUT 1393 00:56:19,360 --> 00:56:21,320 THIS HAS RESPONDED VERY WELL. 1394 00:56:21,320 --> 00:56:24,400 WE HAVE 1 MORE QUESTION HERE. 1395 00:56:24,400 --> 00:56:25,760 SO [INDISCERNIBLE] SAYS, YOU 1396 00:56:25,760 --> 00:56:27,200 PROPOSED IRON RESTRICTION AS A 1397 00:56:27,200 --> 00:56:29,080 THERAPEUTIC APPROACH, HOW DO YOU 1398 00:56:29,080 --> 00:56:34,200 PROPOSE TO FINE BALANCE THIS IN 1399 00:56:34,200 --> 00:56:37,000 TERMS OF ADEQUATE RESTRICTION 1400 00:56:37,000 --> 00:56:37,440 AND IRON DEFICIENCY. 1401 00:56:37,440 --> 00:56:40,400 >> I KIND OF ANSWERED THAT, WE 1402 00:56:40,400 --> 00:56:41,440 ACCOMPLISHED THAT IN OUR 1403 00:56:41,440 --> 00:56:42,200 PATIENTS, THEY WERE NOT 1404 00:56:42,200 --> 00:56:43,160 SIGNIFICANTLY ANEMIC AT THE TIME 1405 00:56:43,160 --> 00:56:45,320 WE WERE ABLE TO IMPACT THIS, 1406 00:56:45,320 --> 00:56:49,840 THAT HAS DIFFERENT KINETICS 1407 00:56:49,840 --> 00:56:50,640 BECAUSE IT'S EXCESS REGULATION 1408 00:56:50,640 --> 00:56:53,920 OR YOU WANT TO MAKE IT MORMAL OR 1409 00:56:53,920 --> 00:56:54,520 EXCESS PRECURSOR ACCUMULATION 1410 00:56:54,520 --> 00:56:56,080 AND I THINK THE REAL ANSWER HERE 1411 00:56:56,080 --> 00:56:58,520 IS THAT YOU SHOULD BE ABLE TO 1412 00:56:58,520 --> 00:57:01,080 ACCOMPLISH THIS AT RELATIVELY 1413 00:57:01,080 --> 00:57:04,800 NORMAL OR JUST LOWER THAN NORMAL 1414 00:57:04,800 --> 00:57:07,240 IRON LEVELS BUT NOT 1S THAT ARE 1415 00:57:07,240 --> 00:57:09,240 LOW AND CAUSING IRON DEFICIENCY 1416 00:57:09,240 --> 00:57:12,680 ANEMIA FOR A VERY SEVERE 1417 00:57:12,680 --> 00:57:14,280 INEFFECTIVE ERYTHROPOIESIS SO 1418 00:57:14,280 --> 00:57:15,720 THE CLINICAL END POINT IS BETTER 1419 00:57:15,720 --> 00:57:19,320 AND IF 1 ADDS THAT 1 HAS EXTRA 1420 00:57:19,320 --> 00:57:22,040 ADVANTAGE OF KEYLATING CELLS 1421 00:57:22,040 --> 00:57:23,320 FREF RENTIALLY BY USING SMALL 1422 00:57:23,320 --> 00:57:28,400 MOLECULES THAT THAT MIGHT 1423 00:57:28,400 --> 00:57:29,640 BE--THAT MIGHT HELP THAT BALANCE 1424 00:57:29,640 --> 00:57:29,880 AS WELL. 1425 00:57:29,880 --> 00:57:31,520 SO I DON'T THINK WE'RE THAT 1426 00:57:31,520 --> 00:57:35,080 WORRIED ABOUT NEEDING TO DRIVE 1427 00:57:35,080 --> 00:57:37,600 IRON DEFICIENCY TO THAT KIND OF 1428 00:57:37,600 --> 00:57:41,160 LEVEL, TO THERE WOULD BE A 1429 00:57:41,160 --> 00:57:42,000 SYSTEMICALLY PROBLEM CAUSING 1430 00:57:42,000 --> 00:57:43,200 MORE ANEMIA IF IT'S NOT RIGHT OR 1431 00:57:43,200 --> 00:57:45,600 IF YOU HAVE IRON IN VERY, VERY 1432 00:57:45,600 --> 00:57:52,920 LOW LEVEL, THE NEXT THING IN AT 1433 00:57:52,920 --> 00:57:55,000 LEAST THE CLINICAL SETTING WE 1434 00:57:55,000 --> 00:57:56,640 DON'T SEE THAT FOR IRON 1435 00:57:56,640 --> 00:57:58,320 DEFICIENCY OTHER THAN IN LONG 1436 00:57:58,320 --> 00:57:59,360 DISTANCE RUNNERS AND ATHLETES OF 1437 00:57:59,360 --> 00:58:01,360 SOME DISTANCE OR ANOTHER, IN THE 1438 00:58:01,360 --> 00:58:03,480 CLINICAL CELTING THAT IMPACTS 1439 00:58:03,480 --> 00:58:06,520 MUSCLE OR IMPACTS ANYTHING ELSE. 1440 00:58:06,520 --> 00:58:09,440 >> SO JAN, CAN I ASK 1 LAST 1441 00:58:09,440 --> 00:58:09,720 QUESTION. 1442 00:58:09,720 --> 00:58:11,560 SO IN YOUR SITUATION WITH THE 1443 00:58:11,560 --> 00:58:13,320 HEALTHY VERSUS UNHEALTHY PATHWAY 1444 00:58:13,320 --> 00:58:14,560 AND YOUR HYPOTHESIS SO THAT MAY 1445 00:58:14,560 --> 00:58:16,440 BE THERE TO DO WHAT HAPPENS 1446 00:58:16,440 --> 00:58:18,040 UNDER STRESS WHEN HAVE YOU 1447 00:58:18,040 --> 00:58:19,320 INCREASED PROTECTION, HAVE YOU 1448 00:58:19,320 --> 00:58:20,720 LOOKED AT ANY SAMPLES OR DO YOU 1449 00:58:20,720 --> 00:58:24,360 PLAN TO FOR PATIENTS THAT ARE 1450 00:58:24,360 --> 00:58:27,480 ANIMAL MODEL THAT ARE SUDDENLY 1451 00:58:27,480 --> 00:58:29,760 PHLEBOTOMIZING AND IN THAT 1452 00:58:29,760 --> 00:58:30,800 SITUATION DEMAND OF 1453 00:58:30,800 --> 00:58:31,360 ERYTHROPOIESIS. 1454 00:58:31,360 --> 00:58:31,800 >> WE PLAN TO. 1455 00:58:31,800 --> 00:58:33,320 WE HAVEN'T DONE THAT. 1456 00:58:33,320 --> 00:58:37,720 AND WE COULD ACTUALLY LOOK AT 1457 00:58:37,720 --> 00:58:39,360 OTHER PLACES WE HAVE MORE 1458 00:58:39,360 --> 00:58:40,720 EFFECTIVE WITH POETICESIS AS 1459 00:58:40,720 --> 00:58:42,400 WELL AND THAT'S MORE OF A 1460 00:58:42,400 --> 00:58:46,160 CHRONIC STATE AND HAVEN'T REALLY 1461 00:58:46,160 --> 00:58:48,200 DECIDED WHETHER IS THERE'S 2 1462 00:58:48,200 --> 00:58:50,520 ISSUES AT PLAY HERE AND 1 IS 1463 00:58:50,520 --> 00:58:52,920 WHAT DO YOU CHRONICALLY? 1464 00:58:52,920 --> 00:58:55,240 SO WE'RE LOOKING AT CHRONIC 1465 00:58:55,240 --> 00:58:56,320 SITUATIONS, WHETHER DOLLAR'S 1466 00:58:56,320 --> 00:58:59,680 VERY EFFECTIVE PRODUCTION, BUT 1467 00:58:59,680 --> 00:59:00,600 PERIPHERAL DESTRUCTION, MIGHT BE 1468 00:59:00,600 --> 00:59:02,080 A MORE INTERESTING MODEL TO LOOK 1469 00:59:02,080 --> 00:59:05,480 AT BUT WE CAN ALSO CLEARLY 1470 00:59:05,480 --> 00:59:07,120 INDUCE HOMOLYSIS AND ANIMAL 1471 00:59:07,120 --> 00:59:08,720 MODELS AND THEN MANY MODEL 1472 00:59:08,720 --> 00:59:09,280 CITIZEN DALLASCOWBOYS.COM 1473 00:59:09,280 --> 00:59:09,880 DALLASCOWBOYS.COMITIES TO DO 1474 00:59:09,880 --> 00:59:12,320 THAT OR USE A HEMOLYTIC ANIMAL 1475 00:59:12,320 --> 00:59:15,640 MODEL AND THE ANSWER IS YES, AND 1476 00:59:15,640 --> 00:59:17,240 ALSO WE'RE INTERESTED IN HOW TO 1477 00:59:17,240 --> 00:59:19,320 MODULATE IT IN NORMAL CELLS AS A 1478 00:59:19,320 --> 00:59:22,680 FIRST START SINCE THERE'S 24% IS 1479 00:59:22,680 --> 00:59:24,960 A PRETTY BIG NUMBER IN OUR 1480 00:59:24,960 --> 00:59:27,520 CULTURE CONDITIONS THAT WE CAN 1481 00:59:27,520 --> 00:59:27,880 MODULATE. 1482 00:59:27,880 --> 00:59:28,160 >> GREAT. 1483 00:59:28,160 --> 00:59:29,960 >> I SHOULD SAY AN ANSWER TO 1484 00:59:29,960 --> 00:59:32,480 YOUR QUESTION, CINDY THAT THERE 1485 00:59:32,480 --> 00:59:38,880 IS MORE IN EFFECTIVENESS THAT 1486 00:59:38,880 --> 00:59:39,640 WE--THERE'S LESS INEFFECTIVENESS 1487 00:59:39,640 --> 00:59:40,880 AND CULTURE THAN THERE IS IN 1488 00:59:40,880 --> 00:59:43,120 REAL LIFE AND SO THAT ALSO GIVES 1489 00:59:43,120 --> 00:59:44,920 A WINDOW TO KIND OF MODULATE 1490 00:59:44,920 --> 00:59:45,280 THAT. 1491 00:59:45,280 --> 00:59:45,640 >> GREAT. 1492 00:59:45,640 --> 00:59:47,360 I THINK WE ARE RIGHT UP AT 1493 00:59:47,360 --> 00:59:47,720 1:00 O'CLOCK. 1494 00:59:47,720 --> 00:59:50,440 SO I WANT TO THANK EVERYONE FOR 1495 00:59:50,440 --> 00:59:50,720 LISTENING. 1496 00:59:50,720 --> 00:59:53,320 THANK JAN VERY MUCH FOR GIVING 1497 00:59:53,320 --> 00:59:54,120 THE TALK ANDIC TAKEN--THEYING 1498 00:59:54,120 --> 00:59:59,160 THE TIME TO SPEAK TO OUR MRST 1499 00:59:59,160 --> 01:00:02,720 STUDENTS TODAY AND RITA STEVENS 1500 01:00:02,720 --> 01:00:04,280 AND WILLIAMS HELPING FOR THE 1501 01:00:04,280 --> 01:00:05,240 AUDIO VISUAL, AND SO EVERYONE 1502 01:00:05,240 --> 01:00:06,960 HAVE A GREAT DAY AND THANK YOU 1503 01:00:06,960 --> 01:00:07,920 VERY MUCH. 1504 01:00:07,920 --> 01:00:08,440 >> BYE, EVERYONE. 1505 01:00:08,440 --> 01:01:33,560 THANK YOU