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 --> 00:00:46,600 Learn more by visiting us online at http://clinicalcenter.nih.gov 11 00:00:46,600 --> 00:00:48,840 OUR SPEAKER TODAY IS DR. NATALIE 12 00:00:48,840 --> 00:00:52,720 SHAW, DR. SHAW IS A LASKER 13 00:00:52,720 --> 00:00:54,360 CLINICAL RESEARCH SCHOLAR AND 14 00:00:54,360 --> 00:00:57,720 HEAD OF PEDIATRIC 15 00:00:57,720 --> 00:00:59,200 NEUROENDOCRINOLOGY GROUP AT 16 00:00:59,200 --> 00:01:00,320 NATIONAL INSTITUTE OF 17 00:01:00,320 --> 00:01:01,320 ENVIRONMENTAL HEALTH SCIENCE. 18 00:01:01,320 --> 00:01:03,200 DR. SHAW EARNED HER MEDICAL 19 00:01:03,200 --> 00:01:04,840 DEGREE FROM SATE UNIVERSITY NEW 20 00:01:04,840 --> 00:01:06,440 YORK MEDICAL SCHOOL AT BUFFALO 21 00:01:06,440 --> 00:01:08,320 AND MASTERS IN MEDICAL SCIENCE 22 00:01:08,320 --> 00:01:10,240 FROM HARVARD MEDICAL SCHOOL. 23 00:01:10,240 --> 00:01:13,840 SHE COMPLETED HER PEDIATRIC 24 00:01:13,840 --> 00:01:15,720 RESIDENCY AT JOE PEDIATRIC 25 00:01:15,720 --> 00:01:17,680 HOSPITAL IN PITTSBURGH, AND 26 00:01:17,680 --> 00:01:20,440 CLINICAL RESEARCH FELLOWSHIP IN 27 00:01:20,440 --> 00:01:22,720 REPRODUCTIVE END KIN UNIT AT 28 00:01:22,720 --> 00:01:23,280 MASSACHUSETTS GENERAL. 29 00:01:23,280 --> 00:01:25,840 SHE SERVED ATTENDING 30 00:01:25,840 --> 00:01:26,840 PEDIATRICIAN AT BOSTON 31 00:01:26,840 --> 00:01:27,520 CHILDREN'S HOSPITAL UNTIL 32 00:01:27,520 --> 00:01:30,480 RECRUITED TO THE NIH IN 2015 AS 33 00:01:30,480 --> 00:01:33,000 A LASKER CLINICAL RESEARCH 34 00:01:33,000 --> 00:01:33,400 SCHOLAR. 35 00:01:33,400 --> 00:01:35,040 AS A INVESTIGATOR, IN THE 36 00:01:35,040 --> 00:01:37,920 CLINICAL RESEARCH BRANCH OF 37 00:01:37,920 --> 00:01:41,720 NIEHS DR. SHAW STUDIED 38 00:01:41,720 --> 00:01:42,640 ENVIRONMENTAL GENETIC CONTROL 39 00:01:42,640 --> 00:01:44,080 PUBERTAL DEVELOPMENT IF RESEARCH 40 00:01:44,080 --> 00:01:47,200 ON EFFECT OF OBESITY ON PUBERTAL 41 00:01:47,200 --> 00:01:49,280 INITIATION AND DEVELOPMENTAL 42 00:01:49,280 --> 00:01:52,760 TRAJECTORY OF CYCLES THAT FOLLOW 43 00:01:52,760 --> 00:01:54,720 MONARCHY TO ESTABLISHMENT OF 44 00:01:54,720 --> 00:01:58,520 CYCLES IN YOUNG ADULTHOOD. 45 00:01:58,520 --> 00:02:01,640 DR. SHAW CONDUCTS STUDIES IN 46 00:02:01,640 --> 00:02:05,720 HEALTHY PEDIATRIC VOLUNTEERS 47 00:02:05,720 --> 00:02:06,840 COMPLIMENTED IN PHENOTYPIC 48 00:02:06,840 --> 00:02:09,320 STUDIES IN PATIENTS WITH RARE 49 00:02:09,320 --> 00:02:12,200 FORMS OF HYPOGOD INISM AND 50 00:02:12,200 --> 00:02:13,680 PATIENT DERIVED NEURAL STEM 51 00:02:13,680 --> 00:02:14,520 CELLS CELLS. 52 00:02:14,520 --> 00:02:15,760 SHE RECEIVED NUMEROUS 53 00:02:15,760 --> 00:02:17,520 RECOMMENDATIONS TO INCLUDE 54 00:02:17,520 --> 00:02:18,520 ENDOCRINE SOCIETY EARLY 55 00:02:18,520 --> 00:02:19,840 INVESTIGATOR AWARD, AMERICAN 56 00:02:19,840 --> 00:02:23,320 SOCIETY FOR CLINICAL 57 00:02:23,320 --> 00:02:24,080 INVESTIGATION YON PHYSICIAN 58 00:02:24,080 --> 00:02:26,200 SCIENTIST AWARD AND WOMEN IN 59 00:02:26,200 --> 00:02:26,960 ENDOCRINOLOGY INVESTIGATOR 60 00:02:26,960 --> 00:02:27,320 AWARD. 61 00:02:27,320 --> 00:02:30,240 SHE IS BOARD CERTIFIED IN 62 00:02:30,240 --> 00:02:32,600 PEDIATRICS AS MEMBER OF THE 63 00:02:32,600 --> 00:02:35,120 ENDOR ENDOCRINE SOCIETY WOMEN IN 64 00:02:35,120 --> 00:02:36,560 ENDOCRINOLOGY AND PEDIATRIC 65 00:02:36,560 --> 00:02:37,160 ENDOCRINE SOCIETY. 66 00:02:37,160 --> 00:02:40,680 HER TALK TODAY IS ENTITLED SMCHD 67 00:02:40,680 --> 00:02:42,400 1 AND EPIGENETICS. 68 00:02:42,400 --> 00:02:45,320 LESSONS LEARNED FROM PATIENTS 69 00:02:45,320 --> 00:02:47,040 BORN WITHOUT A NOSE. 70 00:02:47,040 --> 00:02:47,880 WELCOME OUR SPEAKER, DR. SHAW. 71 00:02:47,880 --> 00:02:48,720 ALL RIGHT. 72 00:02:48,720 --> 00:02:50,480 THANK YOU AGAIN FOR THE 73 00:02:50,480 --> 00:02:55,560 INVITATION TO SPEAK TODAY. 74 00:02:55,560 --> 00:03:01,840 AS DR. BURR CASH FLO I'M A PEDIATRIC 75 00:03:01,840 --> 00:03:02,840 ENDOCRINOLOGIST AT NIEHS IN 76 00:03:02,840 --> 00:03:04,600 NORTH CAROLINA AND TODAY I WILL 77 00:03:04,600 --> 00:03:06,480 TELL YOU A RECENT EXCITING 78 00:03:06,480 --> 00:03:07,600 GENETIC DISCOVERY THAT STARTED 79 00:03:07,600 --> 00:03:11,480 OUT AS A SMALL SIDE PROJECT WHEN 80 00:03:11,480 --> 00:03:15,640 I WAS AN ENDOCRINOLOGY 81 00:03:15,640 --> 00:03:17,880 FELLOWSHIP BIBUT SINCE BLOSSOMED 82 00:03:17,880 --> 00:03:19,880 TO INTERNATIONAL COLLABORATIVE 83 00:03:19,880 --> 00:03:21,560 PROJECT CROSSING MULTIPLE 84 00:03:21,560 --> 00:03:22,400 MEDICAL DISCIPLINES. 85 00:03:22,400 --> 00:03:24,720 I HAVE NOTHING TO DISCLOSE. 86 00:03:24,720 --> 00:03:28,360 HERE IS TODAY'S LEARNING 87 00:03:28,360 --> 00:03:31,240 OBJECTIVE. 88 00:03:31,240 --> 00:03:33,760 AS WITH MANY MEDICAL 89 00:03:33,760 --> 00:03:36,520 DISCOVERIES, SORRY, I'M TRYING 90 00:03:36,520 --> 00:03:42,720 TO MOVE -- THE PICTURES. 91 00:03:42,720 --> 00:03:43,640 OKAY. 92 00:03:43,640 --> 00:03:46,360 AS WITH MANY MEDICAL DISCOVERIES 93 00:03:46,360 --> 00:03:47,760 THIS STARTED WITH AN INTRIGUING 94 00:03:47,760 --> 00:03:48,280 PATIENT. 95 00:03:48,280 --> 00:03:50,320 THE WOMAN PRESENTED TO THE 96 00:03:50,320 --> 00:03:52,040 REPRODUCTIVE ENDOCRINE UNIT AT 97 00:03:52,040 --> 00:03:54,760 MASS GENERAL HOSPITAL WITH A 98 00:03:54,760 --> 00:03:58,200 CHIEF COMPLAINT OF AMENRRHEA OR 99 00:03:58,200 --> 00:03:59,840 ABSENT MEDICAL CYCLES. SHE WAS 100 00:03:59,840 --> 00:04:01,240 BORN WITH MULTIPLE DEFECTS 101 00:04:01,240 --> 00:04:03,360 INCLUDING ABOUT NORMAL EYES AND 102 00:04:03,360 --> 00:04:05,360 CONDITION CALLED ARYANIA OR 103 00:04:05,360 --> 00:04:06,720 COMPLETE ABSENCE OF EXTERNAL 104 00:04:06,720 --> 00:04:07,360 NOSE. 105 00:04:07,360 --> 00:04:09,440 SHE ALSO REPORTED SHE HAD SOME 106 00:04:09,440 --> 00:04:13,120 TYPE OF HYPOTHALAMIC PITUITARY 107 00:04:13,120 --> 00:04:18,440 -- SO DO THESE TWO CONDITIONS 108 00:04:18,440 --> 00:04:21,240 HAVE ANYTHING TO DO WITH 109 00:04:21,240 --> 00:04:25,280 (INAUDIBLE) LET'S STEM BACK A 110 00:04:25,280 --> 00:04:28,680 MOMENT AND REVIEW THE 111 00:04:28,680 --> 00:04:30,160 GONADOTROPIN RELEASING HORMONE 112 00:04:30,160 --> 00:04:33,240 AT BASE OF THE HYPOTHALAMUS. 113 00:04:33,240 --> 00:04:36,520 GNRH SIMULATES PITUITARY 114 00:04:36,520 --> 00:04:40,040 SECRETION WHICH THEN SIMULATE 115 00:04:40,040 --> 00:04:41,520 GOD INIAL SECRETION OF 116 00:04:41,520 --> 00:04:43,800 TESTOSTERONE AND EXTRA DIAL. 117 00:04:43,800 --> 00:04:46,360 BECAUSE REPRODUCTION CONSUMES SO 118 00:04:46,360 --> 00:04:47,240 MUCH ENERGY PARTICULARLY IN 119 00:04:47,240 --> 00:04:48,800 WOMEN, THE ACCESS IS TIGHTLY 120 00:04:48,800 --> 00:04:52,200 CONTROLLED BY THE HYPOTHALAMUS 121 00:04:52,200 --> 00:04:53,000 INTEGRATING INTERNAL AND 122 00:04:53,000 --> 00:04:54,000 EXTERNAL FACTORS. 123 00:04:54,000 --> 00:04:57,840 FOR EXAMPLE, METABOLIC MUST 124 00:04:57,840 --> 00:04:59,440 INDICATE ENERGY STORES FOR 125 00:04:59,440 --> 00:05:01,000 REPRODUCTION AND ENVIRONMENTAL 126 00:05:01,000 --> 00:05:05,560 QUEUES MUST INDICATE FAVORABLE 127 00:05:05,560 --> 00:05:06,360 ENVIRONMENTAL CONDITIONS. 128 00:05:06,360 --> 00:05:07,680 FOR REPRODUCTION. 129 00:05:07,680 --> 00:05:10,400 NOW, THE REPRODUCTIVE ACCESS HAS 130 00:05:10,400 --> 00:05:12,160 TWO VERY DISTINCT FEATURES. 131 00:05:12,160 --> 00:05:15,880 FIRST IS GNRH MUST BE RELEASED 132 00:05:15,880 --> 00:05:16,480 IN A PULSE FASHION AS SHOWN 133 00:05:16,480 --> 00:05:16,880 HERE. 134 00:05:16,880 --> 00:05:20,280 WE CAN'T MEASURE GNRH BECAUSE OF 135 00:05:20,280 --> 00:05:23,760 SHORT HALF LIFE BUT AS YOU CAN 136 00:05:23,760 --> 00:05:25,680 IT IS ALSO PULSE TILE AND IS A 137 00:05:25,680 --> 00:05:28,040 VALIDATED MARKER OF GNRH 138 00:05:28,040 --> 00:05:30,440 SECESSION THAT WE CAN MEASURE IN 139 00:05:30,440 --> 00:05:31,480 THE PERIPHERAL BLOOD. 140 00:05:31,480 --> 00:05:33,360 THE SECOND UNIQUE FEATURE IS 141 00:05:33,360 --> 00:05:35,680 THAT GNRH NEURONS DON'T STARTS 142 00:05:35,680 --> 00:05:36,960 OUT IN THE BRAIN. 143 00:05:36,960 --> 00:05:38,760 THEY MIGRATE THERE FROM THE 144 00:05:38,760 --> 00:05:40,200 PRIMITIVE NOSE. 145 00:05:40,200 --> 00:05:41,720 SO WHEN WE WALK YOU THROUGH THIS 146 00:05:41,720 --> 00:05:45,760 SLIDE, THIS IS A CARTOON SHOWING 147 00:05:45,760 --> 00:05:48,360 THE -- WE OFFER THE ROOT OF THE 148 00:05:48,360 --> 00:05:50,720 NASAL CAVITY WHICH IS SEPARATED 149 00:05:50,720 --> 00:05:53,600 BY THE BRAIN -- BY THE STRUCTURE 150 00:05:53,600 --> 00:05:54,680 CALLED THE CRIB FORM PLATE WHICH 151 00:05:54,680 --> 00:05:56,240 IS ALSO SHOWN HERE. 152 00:05:56,240 --> 00:05:57,800 VERY EARLY IN DEVELOPMENT THERE 153 00:05:57,800 --> 00:06:00,480 IS AN ECTO DERMAL THICKENING 154 00:06:00,480 --> 00:06:02,680 CALLED THE OLFACTORY PLATEAU 155 00:06:02,680 --> 00:06:04,760 WHICH GAVE RISE TO OLFACTORY 156 00:06:04,760 --> 00:06:07,560 NEURONS AND GNHR NEURONS AND 157 00:06:07,560 --> 00:06:09,160 OLFACTORY NEURONS EXTEND AXONS 158 00:06:09,160 --> 00:06:11,560 THROUGH THE TRIGGER FORM PLATE 159 00:06:11,560 --> 00:06:14,960 TO SYNAPSE ANNUAL OLFACTORY BULB 160 00:06:14,960 --> 00:06:17,080 AND THEY HITCH A RIDE ALONG THE 161 00:06:17,080 --> 00:06:18,960 AXONS TO MIGRATE TO BRAIN AND 162 00:06:18,960 --> 00:06:21,760 REACH FINAL DESTINATION IN THE 163 00:06:21,760 --> 00:06:22,120 HYPOTHALAMUS. 164 00:06:22,120 --> 00:06:25,360 AS THIS IS HAPPENING, THE CRIB 165 00:06:25,360 --> 00:06:29,560 REFORM PLATE IS MORE OF A SWAMPY 166 00:06:29,560 --> 00:06:39,600 MESENCHYME SO WHEN THE BONE -- 167 00:06:39,600 --> 00:06:41,720 YOU CAN SEE PERFORATIONS IN THE 168 00:06:41,720 --> 00:06:42,800 CRIB FORM PLATE WHICH ARE RELICS 169 00:06:42,800 --> 00:06:45,280 OF THE OLFACTORY NERVE PASSAGE. 170 00:06:45,280 --> 00:06:47,360 SO GIVEN THIS EMBRYOLOGY YOU 171 00:06:47,360 --> 00:06:49,600 MIGHT PREDICT A DEFECT IN 172 00:06:49,600 --> 00:06:51,040 OLFACTORY STRUCTURE WOULD ALSO 173 00:06:51,040 --> 00:06:54,280 LEAD TO GNRH DEFICIENCY AND YOU 174 00:06:54,280 --> 00:06:55,720 WOULD BE CORRECT THERE IS A A 175 00:06:55,720 --> 00:06:59,560 NAME FOR THAT, COMMON SYNDROME 176 00:06:59,560 --> 00:07:01,640 DEFINED BY COMBINATION OF 177 00:07:01,640 --> 00:07:03,760 OLFACTORY DEFECTS AND -- 178 00:07:03,760 --> 00:07:05,480 DEFECTS. 179 00:07:05,480 --> 00:07:08,240 THESE PATIENTS HAVE ABSENT OR 180 00:07:08,240 --> 00:07:12,040 HYPOPLASTIC OLFACTORY BULKS IN 181 00:07:12,040 --> 00:07:15,120 MRI AND ABSENT SENSE OF SMELL 182 00:07:15,120 --> 00:07:22,520 DIAGNOSED BY A SIMPLE -- SHOWN 183 00:07:22,520 --> 00:07:22,720 HERE. 184 00:07:22,720 --> 00:07:26,120 THE MALES ARE BORN WITH 185 00:07:26,120 --> 00:07:27,560 MICROPENIS AND ORPHANISM AND 186 00:07:27,560 --> 00:07:28,760 BOTH SEXES HAVE ABSENT PUBERTY 187 00:07:28,760 --> 00:07:30,000 AND INFERTILITY. 188 00:07:30,000 --> 00:07:31,920 IF YOU WERE TO DO A FREQUENT 189 00:07:31,920 --> 00:07:35,280 BLOOD SAMPLING STUDY ON SUE 190 00:07:35,280 --> 00:07:37,080 ADJUDICATION YOU WOULD SEE 191 00:07:37,080 --> 00:07:39,040 EXTREMELY SLOW LH LEVELS HERE 192 00:07:39,040 --> 00:07:42,320 SHOWN ON THE BOTTOM IN CONTRAST 193 00:07:42,320 --> 00:07:49,920 TO THIS -- PATTERN IN PATIENTS 194 00:07:49,920 --> 00:07:51,720 HAVE UNCOVERED MUTATIONS IN MORE 195 00:07:51,720 --> 00:07:53,640 THAN 30 GENES THAT HAVE PROVIDED 196 00:07:53,640 --> 00:07:56,920 CRITICAL INSIGHT INTO 197 00:07:56,920 --> 00:07:58,680 REPRODUCTIVE PHYSIOLOGY IN 198 00:07:58,680 --> 00:07:59,160 HEALTH AND DISEASE. 199 00:07:59,160 --> 00:08:02,200 FOR EXAMPLE, SNPS IN MANY OF 200 00:08:02,200 --> 00:08:04,120 THESE GENES HAVE SHOWN UP IN 201 00:08:04,120 --> 00:08:06,760 GENOME WIDE ASSOCIATION STUDIES 202 00:08:06,760 --> 00:08:10,000 FOR AGENT MONARCHY. 203 00:08:10,000 --> 00:08:12,000 GETTING BACK TO OUR PATIENT SHE 204 00:08:12,000 --> 00:08:17,000 SIMPLY THEN HAVE EXTREME FORM OF 205 00:08:17,000 --> 00:08:18,000 KALLMANN SYNDROME. 206 00:08:18,000 --> 00:08:19,760 LET ME TELL YOU MORE ABOUT THE 207 00:08:19,760 --> 00:08:20,280 PRESENTATION. 208 00:08:20,280 --> 00:08:27,080 SHE WAS BORN WITH ARYANIA O OR 209 00:08:27,080 --> 00:08:30,320 SMALL EYES, CATARACTS AND A HOLE 210 00:08:30,320 --> 00:08:33,520 IN THE IRIS, AT AGE 16 SHE HAD 211 00:08:33,520 --> 00:08:35,040 NO BREAST DEVELOPMENT, NO 212 00:08:35,040 --> 00:08:36,680 MENSTRUAL PERIODS AND HAD SMALL 213 00:08:36,680 --> 00:08:38,920 OVARIES AND UTERUS ON THE 214 00:08:38,920 --> 00:08:39,240 ULTRASOUND. 215 00:08:39,240 --> 00:08:42,560 FREQUENT BLOOD SAMPLING 216 00:08:42,560 --> 00:08:44,880 DEMONSTRATED HYPOGONADSISM WITH 217 00:08:44,880 --> 00:08:47,760 UNDETECTABLE LH AND FSH LEVEL 218 00:08:47,760 --> 00:08:51,320 BUS REMAINDER OF HER 219 00:08:51,320 --> 00:08:52,800 HYPOTHALAMIC PITUITARY AXIS WAS 220 00:08:52,800 --> 00:08:53,480 INTACT. 221 00:08:53,480 --> 00:08:55,640 AT AGE 23 SHE HAD A FORMAL 222 00:08:55,640 --> 00:08:58,000 OLFACTORY EVALUATION WHICH 223 00:08:58,000 --> 00:09:00,320 REVEALED COMPLETE INOSINEIA AND 224 00:09:00,320 --> 00:09:02,240 ABSENCE OF OLFACTORY BULBS ON 225 00:09:02,240 --> 00:09:02,640 MRI. 226 00:09:02,640 --> 00:09:04,920 THIS PATIENT WAS ALSO STUDIED IN 227 00:09:04,920 --> 00:09:07,160 COLLABORATION WITH JANICE LEE AT 228 00:09:07,160 --> 00:09:07,520 NIDCR. 229 00:09:07,520 --> 00:09:09,840 AND SHE CAME TO THE NIH CLINICAL 230 00:09:09,840 --> 00:09:12,280 CENTER TO UNDERGO SOMETHING 231 00:09:12,280 --> 00:09:13,840 CALLED THE CONE BEAM CT WHICH 232 00:09:13,840 --> 00:09:15,400 PROVIDES HIGH RESOLUTION IMAGES 233 00:09:15,400 --> 00:09:17,200 OF THE HEART TISSUE STRUCTURES 234 00:09:17,200 --> 00:09:19,680 OF THE FACE AND JAW. 235 00:09:19,680 --> 00:09:22,320 THIS IS SHOWING ENDOCRINE 236 00:09:22,320 --> 00:09:23,720 CONTROL PATIENT AND YOU CAN SEE 237 00:09:23,720 --> 00:09:28,320 THE WELL AIR ATED SINUSES AND 238 00:09:28,320 --> 00:09:29,760 SIGH ANY ORANGE ARROWS POINTING 239 00:09:29,760 --> 00:09:40,640 TO PERFORATION -- PLATE. 240 00:09:40,640 --> 00:09:45,080 HERE YOU CAN SEE -- FACTORY 241 00:09:45,080 --> 00:09:46,280 NEVER GNRH NERVE PASSAGE. 242 00:09:46,280 --> 00:09:47,520 GENERAL IT CAN TESTING WAS 243 00:09:47,520 --> 00:09:48,880 NEGATIVE FOR ALL KNOWN COMMON 244 00:09:48,880 --> 00:09:52,560 GENES. 245 00:09:52,560 --> 00:09:54,440 SO AS I DUG INTO THE LITERATURE 246 00:09:54,440 --> 00:09:56,200 BIT MORE TO TRY TO UNDERSTAND 247 00:09:56,200 --> 00:09:58,200 WHAT THIS PATIENT MIGHT HAVE, I 248 00:09:58,200 --> 00:10:00,000 REALIZED THAT SHE HAD A LITTLE 249 00:10:00,000 --> 00:10:04,040 KNOWN SYNDROME CALLED BOSMA 250 00:10:04,040 --> 00:10:05,400 ARYANIA MICROON THAT WILL EMEA 251 00:10:05,400 --> 00:10:05,760 SYNDROME. 252 00:10:05,760 --> 00:10:10,320 THIS WAS DELINEATED BAY 253 00:10:10,320 --> 00:10:11,720 PHYSICIAN AT THE NIH WHAT USED 254 00:10:11,720 --> 00:10:12,880 TO BE CALLED THE NATIONAL 255 00:10:12,880 --> 00:10:14,400 INSTITUTE OF DENTAL RESEARCH. 256 00:10:14,400 --> 00:10:16,560 AND HE IDENTIFIED THIS SYNDROME 257 00:10:16,560 --> 00:10:21,560 AND CAME ACROSS TWO MALE 258 00:10:21,560 --> 00:10:25,160 PATIENTS WITH A TRIAD OF ARYANIA 259 00:10:25,160 --> 00:10:28,720 OCULAR DEFECT AND HYPOGONADSISM, 260 00:10:28,720 --> 00:10:30,640 ARYANIA IS EXCEEDINGLY RARE WITH 261 00:10:30,640 --> 00:10:32,320 FEWER THAN HUNDRED CASES OVER 262 00:10:32,320 --> 00:10:34,240 THE PAST CENTURY. 263 00:10:34,240 --> 00:10:35,800 THE ETIOLOGY WAS UNKNOWN. 264 00:10:35,800 --> 00:10:37,720 BUT THE PRESENCE OF FAMILIAL 265 00:10:37,720 --> 00:10:40,000 CASES AND THE ABSENCE OF ANY 266 00:10:40,000 --> 00:10:42,720 UNIFYING ENVIRONMENTAL EXPOSURE, 267 00:10:42,720 --> 00:10:44,080 SUGGESTED THAT THERE MAY BE A 268 00:10:44,080 --> 00:10:46,960 GENETIC CAUSE. 269 00:10:46,960 --> 00:10:49,720 SO RECOGNIZING THIS TO BE THE AN 270 00:10:49,720 --> 00:10:50,960 EXCITING OPPORTUNITY FOR GENE 271 00:10:50,960 --> 00:10:52,640 DISCOVERY AND DEFINE THE 272 00:10:52,640 --> 00:10:54,320 REPRODUCTIVE PHENOTYPE FURTHER, 273 00:10:54,320 --> 00:10:56,320 I PARTNERED WITH THIS PATIENT 274 00:10:56,320 --> 00:10:58,880 WHO IS A VERY BRIGHT WOMAN AND 275 00:10:58,880 --> 00:11:03,560 ACTUALLY VERY CURIOUS ABOUT HER 276 00:11:03,560 --> 00:11:09,480 CONDITION ADDITIONAL CASES. 277 00:11:09,480 --> 00:11:10,360 THE NEWSPAPER CLIPPING FROM MORE 278 00:11:10,360 --> 00:11:13,080 THAN 20 YEARS AGO DESCRIBING A 279 00:11:13,080 --> 00:11:13,960 LO DAHLMAN WITH THE SAME EXACT 280 00:11:13,960 --> 00:11:17,400 SYNDROME. 281 00:11:17,400 --> 00:11:18,840 HE HAD UNDERGONE REPRODUCTIVE 282 00:11:18,840 --> 00:11:21,000 SURGERY AS WELL BUT THROUGH A 283 00:11:21,000 --> 00:11:22,320 BIT ROLL GOOGLE DETECTIVE WORK, 284 00:11:22,320 --> 00:11:24,840 I WAS ABLE TO LOCATE THIS MAN 285 00:11:24,840 --> 00:11:27,520 WHO HAD SINCE PASSED AWAY BUT 286 00:11:27,520 --> 00:11:30,920 WHO WAS A LAWYER IN ALBANY AND 287 00:11:30,920 --> 00:11:33,560 WHO WAS ACTUALLY ONE OF THE 288 00:11:33,560 --> 00:11:34,920 FIRST OF THE TWO PATIENTS 289 00:11:34,920 --> 00:11:37,120 ORIGINALLY DESCRIBED BY DR. 290 00:11:37,120 --> 00:11:41,840 JAMES BOSMA. 291 00:11:41,840 --> 00:11:45,680 BY NETWORKING WITH COLLEAGUES IN 292 00:11:45,680 --> 00:11:46,720 ENDOCRINOLOGY AND GENETICS AND 293 00:11:46,720 --> 00:11:48,120 CONTACTING AUTHORS OF PREVIOUS 294 00:11:48,120 --> 00:11:49,480 CASE REPORTS AND FINALLY BY 295 00:11:49,480 --> 00:11:52,280 ENLISTING THE HELP OF A MOTHER 296 00:11:52,280 --> 00:11:54,520 WHO RUNS AN ARYANIA FACEBOOK 297 00:11:54,520 --> 00:11:56,800 PAGE I WAS ABLE TO AMASS A 298 00:11:56,800 --> 00:12:00,480 COHORT OF PATIENTS OF ALMOST 45 299 00:12:00,480 --> 00:12:03,680 CASES AND 51 FAMILY MEMBERS. 300 00:12:03,680 --> 00:12:05,680 IN COLLABORATION WITH COLLEAGUES 301 00:12:05,680 --> 00:12:08,320 AT MASS GENERAL WE NEXT 302 00:12:08,320 --> 00:12:12,600 PERFORMED WHOLE EXOME OR WHOLE 303 00:12:12,600 --> 00:12:14,800 GENOME SEQUENCING THE FIRST 21 304 00:12:14,800 --> 00:12:16,800 PATIENTS AND WHEN YOU LOOK FOR 305 00:12:16,800 --> 00:12:18,720 ULTRA RARE NON-SYNONYMOUS 306 00:12:18,720 --> 00:12:21,440 VARIANTS WE FOUND 84% CASES HAD 307 00:12:21,440 --> 00:12:25,760 MUTATION IN THE SAME GENE SMCHD 308 00:12:25,760 --> 00:12:25,920 1. 309 00:12:25,920 --> 00:12:27,880 ALL HETEROZYGOUS MISSENSE 310 00:12:27,880 --> 00:12:30,160 MUTATIONS RESTRICTED TO AXONS 311 00:12:30,160 --> 00:12:33,840 3-13 OF THIS LARGE 48 EXON 312 00:12:33,840 --> 00:12:35,800 PROTEIN -- GENE. 313 00:12:35,800 --> 00:12:37,200 WE THEN PERFORM SOMETHING CALLED 314 00:12:37,200 --> 00:12:39,200 GENE BASED RARE VARIANT BURDEN 315 00:12:39,200 --> 00:12:41,360 TESTING WHERE WE LOOK AT THE 316 00:12:41,360 --> 00:12:43,360 FREQUENCY OF ALL RARE 317 00:12:43,360 --> 00:12:44,880 NON-SYNONYMOUS VARIANTS AND 318 00:12:44,880 --> 00:12:47,560 EVERY -- IN THE GENOME IN OUR 319 00:12:47,560 --> 00:12:50,120 COHORT COMPARED WITH 60,000 320 00:12:50,120 --> 00:12:52,480 CONTROLS IN WHAT USED TO BE 321 00:12:52,480 --> 00:12:55,040 CALLED EXACT IS NOW REFERRED TO 322 00:12:55,040 --> 00:12:56,360 AS NOMAD BUT AS YOU CAN SEE IN 323 00:12:56,360 --> 00:12:58,840 THIS MANHATTAN PLOT THERE WAS 324 00:12:58,840 --> 00:13:01,000 ONLY ONE GENE ON CHROMOSOME 18 325 00:13:01,000 --> 00:13:02,720 HERE TO ACHIEVE GENOME WIDE 326 00:13:02,720 --> 00:13:05,600 SIGNIFICANCE AND THAT AGAIN WAS 327 00:13:05,600 --> 00:13:06,320 SMCHD 1. 328 00:13:06,320 --> 00:13:10,000 SO WE HAD VERY STRONG 329 00:13:10,000 --> 00:13:12,960 STATISTICAL EVIDENCE FOR ROLE OF 330 00:13:12,960 --> 00:13:14,680 SMCHD 1 BUT ONE SLIGHT PROBLEM. 331 00:13:14,680 --> 00:13:18,080 AND THAT WAS THAT MUTATIONS IN 332 00:13:18,080 --> 00:13:19,520 SMCHD 1 MANY SOME CASES 333 00:13:19,520 --> 00:13:21,840 IDENTICAL TO THOSE FOUND IN OUR 334 00:13:21,840 --> 00:13:23,160 COHORT, HAD ALREADY BEEN SHOWN 335 00:13:23,160 --> 00:13:27,240 TO CAUSE A COMPLETELY DIFFERENT 336 00:13:27,240 --> 00:13:29,960 DISORDER, FASTIO SCAUP LOW 337 00:13:29,960 --> 00:13:31,600 MUSCULAR DYSTROPHY TYPE 2. 338 00:13:31,600 --> 00:13:33,680 A DEGENERATIVE MUSCLE DISORDER 339 00:13:33,680 --> 00:13:36,000 WITH AVERAGE ONSET IN THE 20s. 340 00:13:36,000 --> 00:13:38,520 AND IT CAUSES SLOWLY PROGRESSIVE 341 00:13:38,520 --> 00:13:40,000 WEAKNESS THAT PREDOMINANTLY 342 00:13:40,000 --> 00:13:42,120 AFFECTS THE FACE, SHOULDER 343 00:13:42,120 --> 00:13:43,560 GIRDLE AND UPPER ARMS. 344 00:13:43,560 --> 00:13:45,320 THIS IS THE PATIENT DEMON 345 00:13:45,320 --> 00:13:48,120 INVESTIGATING SOMETHING CALLED 346 00:13:48,120 --> 00:13:49,120 WING SCAPULA WHERE THE MUSCLES 347 00:13:49,120 --> 00:13:52,920 THAT NORMALLY ANCHOR THE SCAPULA 348 00:13:52,920 --> 00:13:55,040 TO THE RIB CAGE ARE WEAKENED 349 00:13:55,040 --> 00:13:55,920 LEADING TO THIS WINGED 350 00:13:55,920 --> 00:13:56,960 APPEARANCE. 351 00:13:56,960 --> 00:13:59,120 SO IT IS NOT UNHEARD OF FOR 352 00:13:59,120 --> 00:14:03,560 MUTATIONS IN ONE PART OF PROTEIN 353 00:14:03,560 --> 00:14:05,120 IN ONE CONDITION AND ANOTHER 354 00:14:05,120 --> 00:14:06,960 PART OF THE PROTEIN TO CAUSE A 355 00:14:06,960 --> 00:14:08,480 DIFFERENT CONDITION BE BUT AS I 356 00:14:08,480 --> 00:14:12,560 SHOW YOU FOR FSHD 2 AND ARYANIA 357 00:14:12,560 --> 00:14:14,720 THAT IS NOT THE CASE. 358 00:14:14,720 --> 00:14:15,800 THIS SLIDE SHOUGHS THE DOMAINS 359 00:14:15,800 --> 00:14:20,960 OF SMCHD 1 PROTEIN, THE 48 EXONS 360 00:14:20,960 --> 00:14:22,280 HERE AND THE DIFFERENT MUTATION 361 00:14:22,280 --> 00:14:24,320 TYPES THAT HAVE BEEN IDENTIFIED 362 00:14:24,320 --> 00:14:24,960 IN THE PATIENTS. 363 00:14:24,960 --> 00:14:32,120 SO IN RED IS THE DELETION 364 00:14:32,120 --> 00:14:33,320 NON-SENSE AND SPLICE SITE AND 365 00:14:33,320 --> 00:14:35,440 BLACK IS MISSENSE. 366 00:14:35,440 --> 00:14:40,160 THE SMCHD 1 MUTATION HAS A 367 00:14:40,160 --> 00:14:42,640 UBIQUITIN LIKE DOMAIN AND FSC 368 00:14:42,640 --> 00:14:43,920 TERMINAL A HINGE DOMAIN. 369 00:14:43,920 --> 00:14:48,120 YOU CAN SEE IN FHSD 2 MUTATIONS 370 00:14:48,120 --> 00:14:50,840 ARE EITHER LOSS OF FUNCTION OR 371 00:14:50,840 --> 00:14:51,880 MISSENSE MUTATIONS. 372 00:14:51,880 --> 00:14:53,960 LOSS OF FUNCTION MUTATION SPAN 373 00:14:53,960 --> 00:14:55,600 THE ENTIRE GENE WHEREAS THE 374 00:14:55,600 --> 00:14:57,800 MISSENSE MUTATIONS SEND TO BE 375 00:14:57,800 --> 00:14:59,040 CLUSTERED IN THIS END TERMINAL 376 00:14:59,040 --> 00:14:59,960 REGION. 377 00:14:59,960 --> 00:15:02,280 THAT IS ALSO THE CASE FOR THE 378 00:15:02,280 --> 00:15:04,680 ARYANIA MUTATIONS, THESE 379 00:15:04,680 --> 00:15:07,920 MUTATIONS ARE ENTIRELY MISSENSE 380 00:15:07,920 --> 00:15:10,600 AND THEY ARE RESTRICTED TO EXONS 381 00:15:10,600 --> 00:15:13,480 3-13. 382 00:15:13,480 --> 00:15:16,000 AT LEAST WE IDENTIFIED FIVE 383 00:15:16,000 --> 00:15:17,480 MUTATIONS THAT ARE IDENTICAL IN 384 00:15:17,480 --> 00:15:20,160 THESE TWO CONDITIONS. 385 00:15:20,160 --> 00:15:24,560 SO YOU MIGHT BE ASKING YOURSELF 386 00:15:24,560 --> 00:15:30,120 IS THIS ONE DISEASE OR TWO? 387 00:15:30,120 --> 00:15:33,000 THAT IS DO PATIENTS WITH ARYANIA 388 00:15:33,000 --> 00:15:35,840 GO ON TO DEVELOP MUSCULAR 389 00:15:35,840 --> 00:15:36,480 DYSTROPHY? 390 00:15:36,480 --> 00:15:37,920 THIS HAS BEEN A DIFFICULT 391 00:15:37,920 --> 00:15:39,720 QUESTION TO ANSWER BECAUSE MANY 392 00:15:39,720 --> 00:15:43,880 OF THE CASE REPORTS ARE IN 393 00:15:43,880 --> 00:15:44,840 INCIDENCE FIRST MADE AND 394 00:15:44,840 --> 00:15:46,080 PATIENTS MAYBE LOSS TO 395 00:15:46,080 --> 00:15:46,560 FOLLOW-UP. 396 00:15:46,560 --> 00:15:48,440 THE SECOND IS THE FACIAL 397 00:15:48,440 --> 00:15:50,880 WEAKNESS THAT IS CHARACTERISTIC 398 00:15:50,880 --> 00:15:55,000 OF FHSD TYPE 2 MULTIPLE 399 00:15:55,000 --> 00:15:56,720 RECONSTRUCTIVE SURGERIES. 400 00:15:56,720 --> 00:15:58,920 WHAT ABOUT PATIENTS WITH 401 00:15:58,920 --> 00:16:00,320 MUSCULAR DYSTROPHY, SUS L 402 00:16:00,320 --> 00:16:01,880 DEFECTS IN SMELL AND 403 00:16:01,880 --> 00:16:02,760 REPRODUCTION? 404 00:16:02,760 --> 00:16:05,440 LET ME TELL YOU MORE ABOUT SMCHD 405 00:16:05,440 --> 00:16:11,920 1 BIOLOGY AND FSHD 2. 406 00:16:11,920 --> 00:16:15,280 SO FSHD 1 STANDS STRUCTURAL 407 00:16:15,280 --> 00:16:17,320 MAINTENANCE OF CHROMOSOME HINGE 408 00:16:17,320 --> 00:16:18,720 DOMAIN CONTAINING PROTEIN ONE, 409 00:16:18,720 --> 00:16:20,760 THE MAJOR FUNCTION IS SERVING AS 410 00:16:20,760 --> 00:16:22,240 A EPIGENERAL IT CAN REPRESSER. 411 00:16:22,240 --> 00:16:24,880 AS A REMINDER EPIGENETICS REFERS 412 00:16:24,880 --> 00:16:27,400 TO CHANGES IN GENE EXPRESSION 413 00:16:27,400 --> 00:16:29,000 WITHOUT CHANGES IN DNA SEQUENCE. 414 00:16:29,000 --> 00:16:31,120 AND YOU CAN THINK OF THIS AS 415 00:16:31,120 --> 00:16:32,920 GIVING OUR GENOME SOME AGILITY. 416 00:16:32,920 --> 00:16:36,240 THAT IS THE ABILITY TO FINE TUNE 417 00:16:36,240 --> 00:16:38,240 GENE EXPRESSION IN SPACE OVER 418 00:16:38,240 --> 00:16:40,720 TIME AND IN RESPONSE TO 419 00:16:40,720 --> 00:16:41,880 ENVIRONMENTAL QUEUES. 420 00:16:41,880 --> 00:16:44,120 GENES ARE TURNED OFF BY ONE OF 421 00:16:44,120 --> 00:16:45,800 THREE MAJOR MECHANISMS. 422 00:16:45,800 --> 00:16:52,200 THE FIRST IS DNA METHYLATION, 423 00:16:52,200 --> 00:16:54,200 SECOND IS BY POST TRANSLATIONAL 424 00:16:54,200 --> 00:16:55,720 MODIFICATION OF HISTONE PROTEIN 425 00:16:55,720 --> 00:16:57,400 TAILS AND THE THIRD IS THROUGH 426 00:16:57,400 --> 00:17:00,320 RNA BASED MECHANISMS SUCH AS 427 00:17:00,320 --> 00:17:03,320 LONG RNs. 428 00:17:03,320 --> 00:17:05,600 SMCHD 1 REPRESSES GENES ON THE 429 00:17:05,600 --> 00:17:08,320 INACTIVE X CHROMOSOME, IT ALSO 430 00:17:08,320 --> 00:17:11,320 SILENCES A SUBSET OF AUTOSOMAL 431 00:17:11,320 --> 00:17:12,720 GENES PARTICULARLY THOSE THAT 432 00:17:12,720 --> 00:17:17,000 ARE IMPRINTED OR MONOLITHICALLY 433 00:17:17,000 --> 00:17:18,320 EXPRESSED FOR OTHER REASONS AND 434 00:17:18,320 --> 00:17:20,920 GENES THAT ARE HIDDEN WITHIN 435 00:17:20,920 --> 00:17:22,760 REPETITIVE DNA SEQUENCES. 436 00:17:22,760 --> 00:17:24,120 IT DOES THIS THROUGH ALL OF 437 00:17:24,120 --> 00:17:25,920 THOSE MECHANISMS THAT I SPOKE 438 00:17:25,920 --> 00:17:30,600 ABOUT BY STABILIZING DNA 439 00:17:30,600 --> 00:17:32,600 METHYLATION REPRESSSIVE HISSTONE 440 00:17:32,600 --> 00:17:35,360 MARKS AND NON-CODING RNEs, IT 441 00:17:35,360 --> 00:17:36,520 IS THOUGHT TO CONTRIBUTE TO 442 00:17:36,520 --> 00:17:38,600 HIGHER ORDER CHROMOSOMAL 443 00:17:38,600 --> 00:17:41,360 STRUCTURE SUCH AS TAGS. 444 00:17:41,360 --> 00:17:45,120 SO HOW DO THESE MUTATIONS LEAD 445 00:17:45,120 --> 00:17:48,000 TO MUSCULAR DYSTROPHY? 446 00:17:48,000 --> 00:17:51,320 IN FHDH 2 MUTATIONS ARE NULL OR 447 00:17:51,320 --> 00:17:54,720 MISSENSE MUTATION MUTATIONS IN SMCHD 1. 448 00:17:54,720 --> 00:17:58,120 WHAT HAPPENS IS THAT THE LOSS OF 449 00:17:58,120 --> 00:18:00,040 REPRESSSIVE ACTIVITY HAS A MAJOR 450 00:18:00,040 --> 00:18:02,560 EFFECTNA IS SPECIFIC LOCUST ON 451 00:18:02,560 --> 00:18:06,160 CHROMOSOME 4 AND THAT IS CALLED 452 00:18:06,160 --> 00:18:08,720 THE D 4Z 4 REPEAT ARRAY. 453 00:18:08,720 --> 00:18:10,480 LOSS OF REPRESSSIVE ACTIVITY 454 00:18:10,480 --> 00:18:13,320 LEADS TO ECTOPIC EXPRESSION OF 455 00:18:13,320 --> 00:18:15,120 DOUBLE HOMEOBOX OR TRANSCRIPTS 456 00:18:15,120 --> 00:18:16,520 FACTOR. 457 00:18:16,520 --> 00:18:21,560 VERY EARLY IN DEVELOPMENT, DUX 4 458 00:18:21,560 --> 00:18:24,120 IS TURNED ON AN PLAYS A ROLE IN 459 00:18:24,120 --> 00:18:25,120 ZYGOTIC GENOME ACTIVATION. 460 00:18:25,120 --> 00:18:27,760 THIS IS WHEN THE GENOME OF THE 461 00:18:27,760 --> 00:18:30,320 ZYGOTE OR EMBRYO IS FIRST 462 00:18:30,320 --> 00:18:31,520 ACTIVATED AND MATERNAL mRNA 463 00:18:31,520 --> 00:18:36,000 AND PROTEINS FROM THE OO SITE 464 00:18:36,000 --> 00:18:37,080 ARE CLEARED. 465 00:18:37,080 --> 00:18:38,280 AT SOME POINT IN DEVELOPMENT 466 00:18:38,280 --> 00:18:42,000 THIS REGION BECOMES METHYLATED 467 00:18:42,000 --> 00:18:44,040 SMCHD 1 BINDS AND STABILIZES ITS 468 00:18:44,040 --> 00:18:45,920 IN DUX 4 IS TURNED OFF. 469 00:18:45,920 --> 00:18:49,120 BUT FOR REASONS THAT AREN'T 470 00:18:49,120 --> 00:18:51,640 ENTIRELY CLEAR, IN FHSD 2 471 00:18:51,640 --> 00:18:53,080 SOMETHING GOES WRONG IN MUSCLE 472 00:18:53,080 --> 00:18:53,560 CELLS. 473 00:18:53,560 --> 00:18:57,320 SO THERE'S LOSS OF METHYLATION, 474 00:18:57,320 --> 00:19:00,440 LOSS OF SMCHD 1 REPRESSSIVE 475 00:19:00,440 --> 00:19:02,400 ACTIVITY AND DUX 4 IS PRODUCED 476 00:19:02,400 --> 00:19:04,600 LEADING TO MUSCLE CELL 477 00:19:04,600 --> 00:19:07,520 APOPTOSIS. 478 00:19:07,520 --> 00:19:09,560 SO BACK TO OUR QUESTIONS IS THIS 479 00:19:09,560 --> 00:19:11,320 ONE DISEASE OR TWO? 480 00:19:11,320 --> 00:19:14,680 WE HAVE STUDIED A COHORT OF FHSD 481 00:19:14,680 --> 00:19:16,920 2 PATIENTS AND FOUND NO CRANIAL 482 00:19:16,920 --> 00:19:18,520 FACIAL OLFACTORY OR REPRODUCTIVE 483 00:19:18,520 --> 00:19:20,000 PHENOTYPES. 484 00:19:20,000 --> 00:19:22,240 WE HAVE ALSO STUDIED A COHORT OF 485 00:19:22,240 --> 00:19:23,920 ARYANNA PATIENTS AT THE CLINICAL 486 00:19:23,920 --> 00:19:28,240 CENTER, IN COLLABORATION WITH 487 00:19:28,240 --> 00:19:29,760 CLARKSTON DONOVAN'S GROUP AND 488 00:19:29,760 --> 00:19:33,000 PERFORMED DEEP NEURAL MUSCULAR 489 00:19:33,000 --> 00:19:34,600 PHENOTYPING STUDIES INCLUDING 490 00:19:34,600 --> 00:19:35,960 MUSCLE US TRAY SOUND AND MUSCLE 491 00:19:35,960 --> 00:19:39,680 MRI AND FOUND NO EVIDENCE OF 492 00:19:39,680 --> 00:19:40,120 FHSD 2. 493 00:19:40,120 --> 00:19:41,920 SO DOES THAT MEAN ARYANIA 494 00:19:41,920 --> 00:19:44,240 PATIENTS DON'T MAKE ANY DUX 495 00:19:44,240 --> 00:19:44,600 SCORE? 496 00:19:44,600 --> 00:19:46,120 THAT IS -- THAT QUESTION HAS 497 00:19:46,120 --> 00:19:48,560 BEEN DIFFICULT TO ANSWER. 498 00:19:48,560 --> 00:19:52,800 THE REASON IS THAT DUX 4 EVEN IN 499 00:19:52,800 --> 00:19:54,360 FSHD 2 MUSCLE IS PRESENT AT LOW 500 00:19:54,360 --> 00:19:57,240 LEVELS AT BOTH THE mRNA AND 501 00:19:57,240 --> 00:19:57,720 PROTEIN LEVEL. 502 00:19:57,720 --> 00:20:00,120 BUT THERE'S TWO POTENTIAL WORK 503 00:20:00,120 --> 00:20:00,720 AROUNDS. 504 00:20:00,720 --> 00:20:04,240 THE FIRST IS THAT WE CAN TREAT 505 00:20:04,240 --> 00:20:06,200 PATIENT FIBROBLASTS WITH TWO 506 00:20:06,200 --> 00:20:08,840 DRUGS THAT RELIEVE EPIGENETIC 507 00:20:08,840 --> 00:20:10,120 REPRESSION TO ASK WHETHER 508 00:20:10,120 --> 00:20:13,120 REPRESSION OF DUX 4 IS DIFFERENT 509 00:20:13,120 --> 00:20:15,880 IN PATIENTS WITH ARYANIA, IN 510 00:20:15,880 --> 00:20:17,760 HEALTHY VOLUNTEERS AND IN 511 00:20:17,760 --> 00:20:23,440 PATIENTS WITH FHSD 2, THAT WAS 512 00:20:23,440 --> 00:20:25,760 THE FIRST -- AND HERE YOU CAN 513 00:20:25,760 --> 00:20:28,880 SEE DUX 4 LEVELS IN HUMAN DERMAL 514 00:20:28,880 --> 00:20:30,120 FIBROBLASTS IN EACH OF THE THREE 515 00:20:30,120 --> 00:20:30,360 GROUPS. 516 00:20:30,360 --> 00:20:32,720 YOU CAN SEE ARYANIA PATIENTS 517 00:20:32,720 --> 00:20:35,000 MAKE SOME DUX 4 BUT IT TENDS TO 518 00:20:35,000 --> 00:20:37,600 BE MUCH LESS THAN THE AMOUNT WE 519 00:20:37,600 --> 00:20:41,760 SEE IN PATIENTS WITH FHSD 2. 520 00:20:41,760 --> 00:20:46,720 WE ALSO OBTAINED MUSCLE BIOPSIES 521 00:20:46,720 --> 00:20:49,160 AND DIFFERENTIATED MYOBLASTS 522 00:20:49,160 --> 00:20:50,520 INTO MYOTUBES BECAUSE THIS IS 523 00:20:50,520 --> 00:20:52,640 KNOWN TO INCREASE DUX 4 524 00:20:52,640 --> 00:20:55,640 EXPRESSION AND HERE YOU CAN SEE 525 00:20:55,640 --> 00:20:59,320 THIS SOMETHING DUX 4 IS HIGHLY 526 00:20:59,320 --> 00:21:02,360 EXPRESSED IN MYOSITES FROM FHSD 527 00:21:02,360 --> 00:21:04,720 2 AND LESS SO IN PATIENTS WITH 528 00:21:04,720 --> 00:21:05,200 ARYANIA. 529 00:21:05,200 --> 00:21:07,120 THIS TELLS US THAT THE CONTROL 530 00:21:07,120 --> 00:21:09,520 OF DUX 4 EXPRESSION IS VERY 531 00:21:09,520 --> 00:21:12,280 DIFFERENT IN THESE TWO 532 00:21:12,280 --> 00:21:12,840 CONDITIONS. 533 00:21:12,840 --> 00:21:14,720 AND PATIENTS WITH ARYANIA MAYBE 534 00:21:14,720 --> 00:21:17,480 PROTECTED FROM DEVELOPING FHSD 535 00:21:17,480 --> 00:21:17,760 2. 536 00:21:17,760 --> 00:21:19,440 WHAT DO WE KNOW SO FAR? 537 00:21:19,440 --> 00:21:22,520 THE FIRST IS THAT ARYANIA 538 00:21:22,520 --> 00:21:26,840 APPEARS TO BE EXTREME FORM OF KA 539 00:21:26,840 --> 00:21:27,360 WILLLMANN SYNDROME BUT NO 540 00:21:27,360 --> 00:21:29,200 GENETIC OVERLAP. 541 00:21:29,200 --> 00:21:31,760 WE SEE INSTEAD STATISTICAL 542 00:21:31,760 --> 00:21:35,080 EVIDENCE MISSENSE MUTATIONS IN 543 00:21:35,080 --> 00:21:37,000 SMCHD 1 ARE PRIMARY EARLY DRIVER 544 00:21:37,000 --> 00:21:37,920 OF EARLY ARYANIA. 545 00:21:37,920 --> 00:21:39,680 LOSS OF FUNCTION AND MISSENSE 546 00:21:39,680 --> 00:21:44,440 MUTATIONS IN SMCHD 1 CAUSE FHSD 547 00:21:44,440 --> 00:21:47,680 2 THE DUX 4 MUSCLE TO BE TOXIN 548 00:21:47,680 --> 00:21:50,080 BUT ARYANIA IN 2 DESPITE 549 00:21:50,080 --> 00:21:51,520 MUTATIONAL OVERLAP APPEARED TO 550 00:21:51,520 --> 00:21:52,760 BE MUTUALLY EXCLUSIVE 551 00:21:52,760 --> 00:21:55,720 CONDITIONS. 552 00:21:55,720 --> 00:21:57,760 BEFORE WE JUMP HEAD FIRST ON TO 553 00:21:57,760 --> 00:22:00,760 THE SMCHD 1 BAND WAGON, I NEED 554 00:22:00,760 --> 00:22:02,440 TO SHARE SOME MORE INFORMATION 555 00:22:02,440 --> 00:22:04,640 WITH YOU ABOUT THESE PATIENTS 556 00:22:04,640 --> 00:22:06,600 THAT TELLS US THAT THE GENETIC 557 00:22:06,600 --> 00:22:08,200 ARCHITECTURE OF THESE CONDITIONS 558 00:22:08,200 --> 00:22:11,280 IS ACTUALLY MUCH MORE COMPLEX. 559 00:22:11,280 --> 00:22:13,680 SO THE TRUTH IS THAT NOT 560 00:22:13,680 --> 00:22:15,080 EVERYONE WITH A MISSENSE 561 00:22:15,080 --> 00:22:18,200 MUTATION IN SMCHD 1 HAS ABNORMAL 562 00:22:18,200 --> 00:22:20,200 PHENOTYPE AND THIS IS SOMETHING 563 00:22:20,200 --> 00:22:22,760 CALLED INCOMPLETE PENETRANTS. 564 00:22:22,760 --> 00:22:24,960 HERE YOU CAN SEE ONE MULTIPLEX 565 00:22:24,960 --> 00:22:27,600 FAMILY WHERE THE MOTHER HAS A 566 00:22:27,600 --> 00:22:30,840 MUTATION IN SMCHD 1 YET HAS NO 567 00:22:30,840 --> 00:22:33,400 CRANIAL FACIAL OR REPRODUCTIVE 568 00:22:33,400 --> 00:22:33,920 PHENOTYPE. 569 00:22:33,920 --> 00:22:37,080 SHE PASSED THE VARIANT ON TO TWO 570 00:22:37,080 --> 00:22:39,880 OF HER CHILDREN, THE DAUGHTER 571 00:22:39,880 --> 00:22:42,720 HAS COMPLETE ARYANIA AND HYPOGOD 572 00:22:42,720 --> 00:22:45,400 INISM AND HER SON HAS NASAL 573 00:22:45,400 --> 00:22:48,520 HYPOPLASIA AND HYPOGONADSISM. 574 00:22:48,520 --> 00:22:50,360 SO THIS ALSO DEMONSTRATES 575 00:22:50,360 --> 00:22:52,240 SOMETHING CALLED VARIABLE 576 00:22:52,240 --> 00:22:53,440 EXPRESSIVITY WHERE INDIVIDUALS 577 00:22:53,440 --> 00:22:55,760 WHERE THE SAME MUTATION EXHIBIT 578 00:22:55,760 --> 00:22:57,240 DIFFERENCES IN DISEASE SEVERITY. 579 00:22:57,240 --> 00:22:59,320 AND YOU CAN SEE THAT WITH THIS 580 00:22:59,320 --> 00:23:02,360 SECOND PEDIGREE AS WELL. 581 00:23:02,360 --> 00:23:06,880 SO THIS FATHER WITH NASAL 582 00:23:06,880 --> 00:23:08,720 HYPOPLASIA AND INOSINEIA PASSED 583 00:23:08,720 --> 00:23:10,520 TO ONE DAUGHTER WITH COMPLETE 584 00:23:10,520 --> 00:23:13,520 ARYANIA AND HYPOGOD INISM AND TO 585 00:23:13,520 --> 00:23:14,520 ANOTHER DAUGHTER SHOWN HERE WHO 586 00:23:14,520 --> 00:23:18,720 HAS NASAL HYPOPLASIA. 587 00:23:18,720 --> 00:23:21,040 SO TO SUMMARIZE IN ARYANIA WE 588 00:23:21,040 --> 00:23:23,960 SEE VARIABLE EXPRESSIVITY, 589 00:23:23,960 --> 00:23:25,520 INCOMPLETE PENETRANTS AND 590 00:23:25,520 --> 00:23:28,000 SOMETHING CALLED PEIO TROUGH PI, 591 00:23:28,000 --> 00:23:29,320 INDIVIDUALS WITH THE SAME 592 00:23:29,320 --> 00:23:31,680 MUTATION EXHIBIT COMPLETELY 593 00:23:31,680 --> 00:23:33,120 DISTINCT DISEASES. 594 00:23:33,120 --> 00:23:39,920 AND WHAT THIS TELLS US IS 595 00:23:39,920 --> 00:23:41,720 MUTATIONS ARE NECESSARY BUT NOT 596 00:23:41,720 --> 00:23:45,560 SUFFICIENT TO CAUSE ARHINIA OR 597 00:23:45,560 --> 00:23:47,400 FSHD 2 AND POINTS TO A POTENTIAL 598 00:23:47,400 --> 00:23:49,200 ROLE FOR GENETIC MODIFIERS 599 00:23:49,200 --> 00:23:54,320 ENVIRONMENTAL EXPOSURES OR TO 600 00:23:54,320 --> 00:23:55,920 RANDOMNESS DURING DEVELOPMENT. 601 00:23:55,920 --> 00:23:58,320 SO HOW CAN WE IDENTIFY 602 00:23:58,320 --> 00:23:59,720 MODIFIERS 603 00:23:59,720 --> 00:24:00,240 SO WE WERE 604 00:24:00,240 --> 00:24:02,240 TALKING HOW TO IDENTIFY THESE 605 00:24:02,240 --> 00:24:03,200 GENETIC OR ENVIRONMENTAL 606 00:24:03,200 --> 00:24:04,920 MODIFIERS. 607 00:24:04,920 --> 00:24:05,440 S WHAT WE WERE HOPING TO DO IS A 608 00:24:05,440 --> 00:24:07,360 FSHD VERSUS ARHINIA GWAS STUDY 609 00:24:07,360 --> 00:24:09,120 BUT NOT SURE TO HAVE ENOUGH 610 00:24:09,120 --> 00:24:12,880 PATIENTS TO HAVE STATISTICAL 611 00:24:12,880 --> 00:24:15,040 POWER. 612 00:24:15,040 --> 00:24:20,680 WE NEED TO OPEN A WET LAB. 613 00:24:20,680 --> 00:24:22,320 THAT POSE AS SLIDE PROBLEM SINCE 614 00:24:22,320 --> 00:24:24,440 I HAVEN'T USED A PIPETTE SINCE 615 00:24:24,440 --> 00:24:26,720 MY COLLEGE DAYS BUT FORTUNATELY 616 00:24:26,720 --> 00:24:28,960 MY SCIENTIFIC DIRECTOR DARRELL 617 00:24:28,960 --> 00:24:31,040 ZELDON WAS SUPPORTIVE OF MY 618 00:24:31,040 --> 00:24:32,280 PURSUING THIS WORK AND GAVE ME 619 00:24:32,280 --> 00:24:34,800 THE RESOURCES AND PERSONNEL TO 620 00:24:34,800 --> 00:24:36,080 OPEN A WET LAB SEVERAL YEARS 621 00:24:36,080 --> 00:24:38,720 AGO. 622 00:24:38,720 --> 00:24:41,440 IF WE ARE GOING TO MODEL THESE 623 00:24:41,440 --> 00:24:42,760 MUTATION, THERE'S TWO CRITICAL 624 00:24:42,760 --> 00:24:44,240 QUESTIONS TO ANSWER. 625 00:24:44,240 --> 00:24:46,560 THE FIRST IS WHAT DO MISSENSE 626 00:24:46,560 --> 00:24:48,200 MUTATIONS DO TO SMCHD 1 627 00:24:48,200 --> 00:24:49,760 FUNCTION? 628 00:24:49,760 --> 00:24:52,440 MISSENSE MUTATIONS ARE A BIT 629 00:24:52,440 --> 00:24:54,120 TRICKIER THAN NULL MUTATIONS 630 00:24:54,120 --> 00:24:55,640 THEY COULD MAKE PROTEIN LESS 631 00:24:55,640 --> 00:24:58,840 ACTIVE MORE ACTIVE OR EVEN 632 00:24:58,840 --> 00:25:00,200 GRANTED A NOVEL FUNCTION. 633 00:25:00,200 --> 00:25:01,960 THE SECOND IS WHAT CELL TYPES 634 00:25:01,960 --> 00:25:03,320 SHOULD WE STUDY? 635 00:25:03,320 --> 00:25:05,760 SO WE CAN START WITH A HUMAN 636 00:25:05,760 --> 00:25:07,760 EMBRYONIC STEM CELL, BECAUSE WE 637 00:25:07,760 --> 00:25:10,960 KNOW THIS DEFECTIVE OCCURS EARLY 638 00:25:10,960 --> 00:25:13,400 DURING EMBRYO GENESIS BUT THAT 639 00:25:13,400 --> 00:25:14,960 IS UNLIKELY TO BE THE SPECIFIC 640 00:25:14,960 --> 00:25:16,200 CELL TYPE AFFECTED OR WE WOULD 641 00:25:16,200 --> 00:25:18,000 SEE ADDITIONAL DEFECTS IN OTHER 642 00:25:18,000 --> 00:25:23,160 ORGAN SYSTEMS. 643 00:25:23,160 --> 00:25:25,440 SO TO ANSWER THE FIRST QUESTION, 644 00:25:25,440 --> 00:25:27,240 LET'S GO BACK FOR A MOMENT TO 645 00:25:27,240 --> 00:25:29,800 BASIC BIOLOGY OF THE SMCHD 1 646 00:25:29,800 --> 00:25:31,400 PROTEIN. 647 00:25:31,400 --> 00:25:33,920 AS I MENTIONED, IT IS A LARGE 648 00:25:33,920 --> 00:25:36,240 2005 AMINO ACID PROTEIN WITH 649 00:25:36,240 --> 00:25:38,880 SEVERAL DOMAINS IN THE END 650 00:25:38,880 --> 00:25:42,680 TERMINAL REGION AND SMCHD HINGE 651 00:25:42,680 --> 00:25:44,360 AT THE C TERMINAL REGION. 652 00:25:44,360 --> 00:25:47,920 IN THE CELL IT INTERESTS IN THIS 653 00:25:47,920 --> 00:25:49,080 INTERESTING BARBELL LIKE 654 00:25:49,080 --> 00:25:50,800 CONFIGURATION WITH DIMERIZATION 655 00:25:50,800 --> 00:25:54,040 OF BOTH N AND C TERMINALS. 656 00:25:54,040 --> 00:25:55,080 MY GROUP HAS BEEN VERY 657 00:25:55,080 --> 00:25:56,440 INTERESTED IN THE END TERMINAL 658 00:25:56,440 --> 00:25:58,800 REGION BECAUSE THAT'S WHERE THE 659 00:25:58,800 --> 00:26:00,600 MUTATIONS LIE AND IN 660 00:26:00,600 --> 00:26:03,120 COLLABORATION WITH LARS PETERSON 661 00:26:03,120 --> 00:26:07,640 AT NIEHS WE CRYSTALLIZE A 662 00:26:07,640 --> 00:26:09,560 PROTEIN CONSTRUCT ENCOMPASSING 663 00:26:09,560 --> 00:26:12,680 AMINO ACIDS 24 TO 580 WHICH 664 00:26:12,680 --> 00:26:14,680 DEMONSTRATES ATP DEPENDENT 665 00:26:14,680 --> 00:26:16,600 DIMERIZATION, THIS IS THE ATP 666 00:26:16,600 --> 00:26:19,640 MOLECULE SHOWN HERE. 667 00:26:19,640 --> 00:26:22,560 THE DIMER STABILIZED BY THIS 668 00:26:22,560 --> 00:26:24,160 SWATHING OF THIS STRAP IN PURPLE 669 00:26:24,160 --> 00:26:27,840 AND UBIQUITIN LIKE DOMAIN. 670 00:26:27,840 --> 00:26:28,640 SHOWN IN BEIGE. 671 00:26:28,640 --> 00:26:31,880 IF WE LOOK AT THE MUTATIONS THE 672 00:26:31,880 --> 00:26:34,760 ARHINIA MUTATIONS ARE IN PINK, 673 00:26:34,760 --> 00:26:36,840 FSHD IN BLUE AND MUTATION SEEN 674 00:26:36,840 --> 00:26:38,160 IN BOTH CONDITIONS ARE IN 675 00:26:38,160 --> 00:26:39,160 ORANGE. 676 00:26:39,160 --> 00:26:41,680 AND YOU CAN SEE THAT THE 677 00:26:41,680 --> 00:26:43,720 MUTATIONS IN BOTH CONDITIONS ARE 678 00:26:43,720 --> 00:26:46,040 CLUSTERED AROUND THE ATP ACE 679 00:26:46,040 --> 00:26:47,200 DOMAIN OR AT THE DIMER 680 00:26:47,200 --> 00:26:49,120 INTERFACE. 681 00:26:49,120 --> 00:26:51,440 HOW DO THESE MUTATIONS AFFECT 682 00:26:51,440 --> 00:26:53,000 PROTEIN FUNCTION? 683 00:26:53,000 --> 00:26:54,240 WE LOOKED AT TWO DIFFERENT 684 00:26:54,240 --> 00:26:58,320 FUNCTIONS ATP ACE ACTIVITY AND 685 00:26:58,320 --> 00:26:59,240 DIMERIZATION. 686 00:26:59,240 --> 00:27:01,640 SHOWN HERE ARE ALL OF THE 687 00:27:01,640 --> 00:27:03,560 ARHINIA MUTANT CONSTRUCTS, WITH 688 00:27:03,560 --> 00:27:06,600 THE EXCEPTION OF E 147A WHICH IS 689 00:27:06,600 --> 00:27:10,720 OUR POSITIVE CONTROL WHICH NOW 690 00:27:10,720 --> 00:27:13,080 ABOLISHES ATP ACE ACTIVITY. 691 00:27:13,080 --> 00:27:14,360 AND THREE FSHD MUTATIONS. 692 00:27:14,360 --> 00:27:16,960 AND YOU CAN SEE THAT THERE IS -- 693 00:27:16,960 --> 00:27:20,280 THERE ARE VERY MINOR EFFECTS ON 694 00:27:20,280 --> 00:27:21,840 ATPASE ACTIVITY AND THERE 695 00:27:21,840 --> 00:27:24,120 DOESN'T SEEM TO BE AN EFFECT IN 696 00:27:24,120 --> 00:27:25,800 ONE DIRECTION AND ONE CONDITION 697 00:27:25,800 --> 00:27:27,080 AND OPPOSITE EFFECT IN THE OTHER 698 00:27:27,080 --> 00:27:30,360 CONDITION. 699 00:27:30,360 --> 00:27:31,720 CONTRAST THAT WITH DIMERIZATION 700 00:27:31,720 --> 00:27:33,600 WITH WHERE IN ARHINIA WE SEE NO 701 00:27:33,600 --> 00:27:36,720 EFFECT ON DIMERIZATION, WHEREAS 702 00:27:36,720 --> 00:27:39,400 ALL THREE FSHD 2 MUTATIONS LED 703 00:27:39,400 --> 00:27:42,960 TO LOSS OF DIMERIZATION. 704 00:27:42,960 --> 00:27:46,520 WHAT ABOUT LESSONS FROM SMCHD 1 705 00:27:46,520 --> 00:27:48,040 MUTANT ANIMAL MODELS? 706 00:27:48,040 --> 00:27:51,720 IN COLLABORATION WITH MIKO AND 707 00:27:51,720 --> 00:27:56,160 ERICA DAVIS FORMERLY AT DUKE WE 708 00:27:56,160 --> 00:27:59,240 SUPPRESSED SMCHD 1 IN ZEBRAFISH 709 00:27:59,240 --> 00:28:01,400 AND SAW CHANGES IN FACIAL 710 00:28:01,400 --> 00:28:02,640 CARTILAGE CAVITY AND SMALLER 711 00:28:02,640 --> 00:28:05,640 EYES. HOWEVER A COMPETING GROUP 712 00:28:05,640 --> 00:28:07,640 OVEREXPRESSED SMCHD 1 TO MODEL A 713 00:28:07,640 --> 00:28:09,640 GAIN OF FUNCTION MECHANISM AND 714 00:28:09,640 --> 00:28:13,960 ALSO EXERTS SMALLER EYES. 715 00:28:13,960 --> 00:28:17,480 SMCHD 1 NULL MOUSE BUZZ REPORTED 716 00:28:17,480 --> 00:28:20,360 IN 2005 BY LOUIS'S GROUP AND 717 00:28:20,360 --> 00:28:23,240 INTERESTINGLY THESE MICE DO NOT 718 00:28:23,240 --> 00:28:25,160 HAVE FSHD 2 OR ARHINIA, PERHAPS 719 00:28:25,160 --> 00:28:26,840 BECAUSE THE MOUSE DOESN'T 720 00:28:26,840 --> 00:28:30,600 EXPRESS THE HUMAN FORM OF DUX 4. 721 00:28:30,600 --> 00:28:32,680 THE ONLY PHENOTYPE THAT WE HAVE 722 00:28:32,680 --> 00:28:35,680 SEEN IS THAT THERE WAS INCREASE 723 00:28:35,680 --> 00:28:37,240 EMBRYONICALLY LETHALITY IN 724 00:28:37,240 --> 00:28:40,400 FEMALES LIKELY BECAUSE OF SMCHD 725 00:28:40,400 --> 00:28:42,640 1 ROLE IN EXON ACTIVATION. 726 00:28:42,640 --> 00:28:47,000 WE GENERATED AN SMCHD 1 KNOCK IN 727 00:28:47,000 --> 00:28:49,240 MOUSE USING MUTATION WE SEE MOST 728 00:28:49,240 --> 00:28:52,240 COMMONLY IN OUR COHORT, L 141F. 729 00:28:52,240 --> 00:28:55,040 AND THESE MICE APPEAR TO 730 00:28:55,040 --> 00:28:56,680 PHENOCOPY THE NULL MICE, 731 00:28:56,680 --> 00:28:58,800 SUGGESTING A LOSS OF FUNCTION 732 00:28:58,800 --> 00:29:02,880 MECHANISM IN ARHINIA. 733 00:29:02,880 --> 00:29:04,320 THE STRONGEST EVIDENCE THESE 734 00:29:04,320 --> 00:29:05,240 MUTATIONS MIGHT BE BEHAVING IN 735 00:29:05,240 --> 00:29:07,120 THE A LOSS OF FUNCTION FASHION 736 00:29:07,120 --> 00:29:09,240 CAME WHEN WE WENT BACK TO OUR 737 00:29:09,240 --> 00:29:11,120 PATIENTS, AND USED A CLINICAL 738 00:29:11,120 --> 00:29:14,000 TEST THAT USED -- IS USED TO 739 00:29:14,000 --> 00:29:15,640 DIAGNOSE FSHD 2 AND THAT'S TO 740 00:29:15,640 --> 00:29:17,680 LOOK AT THE METHYLATION LEVEL AT 741 00:29:17,680 --> 00:29:21,760 THE DUX 4 PROMOTER AT THAT D 4Z 742 00:29:21,760 --> 00:29:25,040 4 REGION. 743 00:29:25,040 --> 00:29:27,880 IN FSHD 2 THOSE PATIENTS HAD 744 00:29:27,880 --> 00:29:29,720 LESS THAN 25% METHYLATION. 745 00:29:29,720 --> 00:29:31,840 AND WHAT WE SAW IS THAT THE 746 00:29:31,840 --> 00:29:34,920 MAJORITY OF OUR PATIENTS WITH 747 00:29:34,920 --> 00:29:37,360 SMCHD 1 MUTATION ALSO 748 00:29:37,360 --> 00:29:39,000 DEMONSTRATED HYPOMETHYLATION, 749 00:29:39,000 --> 00:29:40,240 UNAFFECTED FAMILY MEMBERS 750 00:29:40,240 --> 00:29:43,720 WITHOUT THE MUTATION DID NOT. 751 00:29:43,720 --> 00:29:46,920 SO IF WE GO BACK TO OUR 752 00:29:46,920 --> 00:29:48,320 QUESTION, FIRST QUESTION, IT 753 00:29:48,320 --> 00:29:49,920 SEEMS SAFE TO MODEL THESE 754 00:29:49,920 --> 00:29:51,240 MUTATIONS AS A LOSS OF FUNCTION 755 00:29:51,240 --> 00:29:52,800 MECHANISM. 756 00:29:52,800 --> 00:29:54,280 BUT THAT LEAVES US WITH THE 757 00:29:54,280 --> 00:29:55,640 SECOND QUESTION OF WHAT CELL 758 00:29:55,640 --> 00:29:57,600 TYPE WE SHOULD BE STUDYING. 759 00:29:57,600 --> 00:30:00,000 SO IF YOU RECALL THE CLINICAL 760 00:30:00,000 --> 00:30:01,720 TRIAD OF THIS CONDITION, THAT IT 761 00:30:01,720 --> 00:30:04,680 AFFECTS THE NOSE, EYES, AND 762 00:30:04,680 --> 00:30:07,120 LEADS TO GNRH DEFICIENCY, THAT 763 00:30:07,120 --> 00:30:10,800 SHOULD TAKE US BACK TO EARLY 764 00:30:10,800 --> 00:30:12,640 EMBRYONIC DEVELOPMENT AROUND 765 00:30:12,640 --> 00:30:15,000 WEEK 3 OR 4 AND HERE I'M SHOWING 766 00:30:15,000 --> 00:30:17,440 THE DEVELOPING NEURAL PLATE 767 00:30:17,440 --> 00:30:20,080 WHICH WILL -- IN PINK WHICH WILL 768 00:30:20,080 --> 00:30:21,920 INVAGINATE AND FORM THE NEURAL 769 00:30:21,920 --> 00:30:24,640 TUBE TO GIVE RISE TO THE BRAIN 770 00:30:24,640 --> 00:30:25,040 AND SPINAL CHORD. 771 00:30:25,040 --> 00:30:27,240 BUT THERE ARE THESE TWO CELLULAR 772 00:30:27,240 --> 00:30:30,120 POPULATIONS RIGHT AT THE NEURAL 773 00:30:30,120 --> 00:30:32,400 PLATE BORDER, THE NEURAL CREST 774 00:30:32,400 --> 00:30:35,440 CELLS AND CRANIO CELLS. 775 00:30:35,440 --> 00:30:35,960 NEURAL CREST CELLS MIGRATE 776 00:30:35,960 --> 00:30:37,040 THROUGHOUT THE BODY GIVING RISE 777 00:30:37,040 --> 00:30:40,160 TO DIVERSE ORGANS SUCH AS THE 778 00:30:40,160 --> 00:30:43,120 HEART AND GANGLIA AND 779 00:30:43,120 --> 00:30:44,200 MELANOCYTES. 780 00:30:44,200 --> 00:30:47,440 HOWEVER THE CRANIAL SUBDIVISION 781 00:30:47,440 --> 00:30:49,480 OF NEURAL CREST CELL GAVE RISE 782 00:30:49,480 --> 00:30:52,800 TO FACIAL SKELETON, PERIOCULAR 783 00:30:52,800 --> 00:30:55,680 MESENCHYME AND OLFACTORY NERVE 784 00:30:55,680 --> 00:31:00,040 AND SHEATHING CELLS. 785 00:31:00,040 --> 00:31:02,120 THE CRANIAL CELLS ARE PRIMITIVE 786 00:31:02,120 --> 00:31:03,480 SENSORY ORGANS AND WE ALREADY 787 00:31:03,480 --> 00:31:07,200 TALKED ABOUT HOW THE OLFACTORY 788 00:31:07,200 --> 00:31:10,080 PLAQUE CODE GAVE RISE TO GNRH 789 00:31:10,080 --> 00:31:11,840 NEURONS, OLFACTORY NEURONS AND 790 00:31:11,840 --> 00:31:13,560 ALSO RELEASES GROWTH FACTORS 791 00:31:13,560 --> 00:31:15,240 THAT INDUCE FORMATION OF THE 792 00:31:15,240 --> 00:31:16,280 NASAL SKELETON. 793 00:31:16,280 --> 00:31:21,600 SO JUST AS A REMINDER, THE GNRH 794 00:31:21,600 --> 00:31:22,840 NEURONS HITCH ALONG THESE 795 00:31:22,840 --> 00:31:24,320 OLFACTORY NERVE AXONS TO THE 796 00:31:24,320 --> 00:31:28,120 BRAIN AND THESE ARE BLAH CODE 797 00:31:28,120 --> 00:31:30,440 DERIVATIVES BUT THE SHEATHING 798 00:31:30,440 --> 00:31:32,080 CELLS WHICH IS CRITICAL FOR THE 799 00:31:32,080 --> 00:31:34,640 INTEGRITY OF OLFACTORY NERVES 800 00:31:34,640 --> 00:31:36,680 ARE DERIVATIVES OF CRANIAL 801 00:31:36,680 --> 00:31:37,720 NEURAL CREST CELLS SO WE NEED TO 802 00:31:37,720 --> 00:31:38,840 CONSIDER BOTH CELLULAR 803 00:31:38,840 --> 00:31:39,440 POPULATIONS. 804 00:31:39,440 --> 00:31:41,400 AND IN FACT, WHEN WE GO TO OUR 805 00:31:41,400 --> 00:31:43,640 ANIMAL MODELS BESEE EXPRESSION 806 00:31:43,640 --> 00:31:48,040 IN BOTH OF THESE CELL TYPES SO 807 00:31:48,040 --> 00:31:51,360 THIS IS XGAL STAINING OF EMBRYOS 808 00:31:51,360 --> 00:31:53,560 AT E 9.5 AND YOU CAN SEE DEEP 809 00:31:53,560 --> 00:31:59,240 STAINING AT THE NASAL PLA CODE, 810 00:31:59,240 --> 00:32:02,240 THIS IS SHOWING XENOPUS EMBRYOS 811 00:32:02,240 --> 00:32:06,000 STAINING ON THE EYES, THE NASAL 812 00:32:06,000 --> 00:32:08,000 PLACODES AND FOUR MIGRATORY 813 00:32:08,000 --> 00:32:10,960 STREAMS OF NEURAL CREST CELLS. 814 00:32:10,960 --> 00:32:14,240 SO WE HAVE GOT OUR PATIENTS, AND 815 00:32:14,240 --> 00:32:16,360 OUR CONTROLS AND SO WE OBTAIN 816 00:32:16,360 --> 00:32:21,120 SKIN BIOPSIES AND CREATED INDUCE 817 00:32:21,120 --> 00:32:22,680 PLURIPOTENT STEM CELLS FROM BOTH 818 00:32:22,680 --> 00:32:24,320 PATIENTS AND CONTROLS. 819 00:32:24,320 --> 00:32:27,320 AND WE ALSO STUDIED THE H 9 820 00:32:27,320 --> 00:32:29,080 HUMAN EMBRYONIC STEM CELL LINE 821 00:32:29,080 --> 00:32:30,720 BECAUSE IN THIS LINE WE WERE 822 00:32:30,720 --> 00:32:34,880 ABLE TO KNOCK OUT SMCHD 1. 823 00:32:34,880 --> 00:32:37,160 AND THESE -- THIS ES CELL MODEL 824 00:32:37,160 --> 00:32:38,480 IS IMPORTANT BECAUSE THESE CELLS 825 00:32:38,480 --> 00:32:40,240 ARE ISOGENIC, MEANING THEY HAVE 826 00:32:40,240 --> 00:32:42,120 THE SAME GENETIC BACKGROUND. 827 00:32:42,120 --> 00:32:44,600 SO IT TAKES AWAY SOME OF THE 828 00:32:44,600 --> 00:32:46,640 VARIABILITY THAT WE WOULD SEE IN 829 00:32:46,640 --> 00:32:48,040 OUR PATIENT CELL LINE SO WE USE 830 00:32:48,040 --> 00:32:49,640 BOTH OF THESE FOR OUR APPROACH. 831 00:32:49,640 --> 00:32:51,840 THIS IS JUST SHOWING A WESTERN 832 00:32:51,840 --> 00:32:53,560 BLOT WITH SUCCESSFUL KNOCK OUT 833 00:32:53,560 --> 00:32:56,960 OF SMCHD 1 IN H 9. 834 00:32:56,960 --> 00:32:58,840 I DON'T HAVE TIME TO GO INTO ALL 835 00:32:58,840 --> 00:33:00,200 THE DETAILS BUT I WILL TELL YOU 836 00:33:00,200 --> 00:33:03,600 THAT WE STARTED BY DERIVING 837 00:33:03,600 --> 00:33:05,120 NEURAL CREST CELLS FROM THESE 838 00:33:05,120 --> 00:33:07,640 IPS CELLS AND THE H 9 LINE AND 839 00:33:07,640 --> 00:33:09,240 SAW NO ABNORMAL PHENOTYPE. 840 00:33:09,240 --> 00:33:11,360 THERE WAS NORMAL PROLIFERATION 841 00:33:11,360 --> 00:33:13,480 AND NORMAL MIGRATION. 842 00:33:13,480 --> 00:33:18,160 SO WE MOVED ON AND USED AND 843 00:33:18,160 --> 00:33:20,120 ESTABLISHED CRANIAL PLA CODE 844 00:33:20,120 --> 00:33:21,480 PROTOCOL THAT INVOLVES DUAL 845 00:33:21,480 --> 00:33:24,720 INHIBITION OF TGF BETA AND BMP, 846 00:33:24,720 --> 00:33:26,720 THIS TAKES 11 DAYS. 847 00:33:26,720 --> 00:33:30,040 WHAT WE SAW RIGHT AWAY, IS 848 00:33:30,040 --> 00:33:30,960 SIGNIFICANT CELL DEATH. 849 00:33:30,960 --> 00:33:32,960 SO THIS IS CRYSTAL VIOLET 850 00:33:32,960 --> 00:33:35,640 STAINING IN TWO OF OUR WILD TYPE 851 00:33:35,640 --> 00:33:37,520 ES CELLS AND IN OUR KNOCK 852 00:33:37,520 --> 00:33:39,640 OUTLINE. 853 00:33:39,640 --> 00:33:42,320 AND YOU CAN SEE PURPLE MEANS 854 00:33:42,320 --> 00:33:45,240 CELLS ARE ALIVE AND WHITE 855 00:33:45,240 --> 00:33:46,760 INDICATES CELLS DEATH SO WE SAW 856 00:33:46,760 --> 00:33:51,160 THIS AROUND DAY 3 TO DAY 4. 857 00:33:51,160 --> 00:33:53,040 WHAT WAS UPREGULATED AT THIS 858 00:33:53,040 --> 00:33:53,520 TIME? 859 00:33:53,520 --> 00:33:55,680 NONE OTHER THAN THE DUX 4 MUZZLE 860 00:33:55,680 --> 00:33:58,960 TOXIN WE TALKED ABOUT IN FHSD 2. 861 00:33:58,960 --> 00:34:00,400 THIS IS SHOWING INCREASE 862 00:34:00,400 --> 00:34:02,520 BEGINNING AT DAY 2 AT THE mRNA 863 00:34:02,520 --> 00:34:04,600 LEVEL AND ROBUST EXPRESSION AT 864 00:34:04,600 --> 00:34:08,200 THE PROTEIN LEVEL AT DAYS FOUR 865 00:34:08,200 --> 00:34:10,720 AND FIVE WE ALSO PERFORMED A 866 00:34:10,720 --> 00:34:12,040 TIME COURSE RNA SEQ EXPERIMENT 867 00:34:12,040 --> 00:34:14,200 AND YOU CAN SEE IN THIS VOLCANO 868 00:34:14,200 --> 00:34:16,440 PLOT THAT MANY GENES WERE 869 00:34:16,440 --> 00:34:18,640 DOWN-REGULATED SHOWN IN GREEN IN 870 00:34:18,640 --> 00:34:20,080 OUR KNOCK OUT CELLS AND MANY 871 00:34:20,080 --> 00:34:21,600 WERE UPREGULATED SHOWN IN RED. 872 00:34:21,600 --> 00:34:25,240 BUT ALL OF THESE GENES THAT ARE 873 00:34:25,240 --> 00:34:27,240 LISTED HERE ARE KNOWN DUX 4 874 00:34:27,240 --> 00:34:27,640 TARGETS. 875 00:34:27,640 --> 00:34:31,080 AND I WILL JUST HIGHLIGHT ONE OF 876 00:34:31,080 --> 00:34:32,360 THEM, TRIM 43 THAT WE WILL COME 877 00:34:32,360 --> 00:34:33,400 BACK TO. 878 00:34:33,400 --> 00:34:35,680 SO THE NEXT THING WE DID WAS TO 879 00:34:35,680 --> 00:34:37,600 DEMONSTRATE THAT THE CELLULAR 880 00:34:37,600 --> 00:34:38,760 PHENOTYPE WAS SPECIFICALLY 881 00:34:38,760 --> 00:34:41,840 CAUSED BY LOSS OF SMCHD 1. 882 00:34:41,840 --> 00:34:44,640 WE DID THAT BY KNOCKING IN OR 883 00:34:44,640 --> 00:34:47,000 REINTRODUCING SMCHD 1 SHOWN HERE 884 00:34:47,000 --> 00:34:50,800 ARE THE KNOCK OUTLINES WITH 885 00:34:50,800 --> 00:34:53,120 INCREASE DUX 4 EXPRESSION NO 886 00:34:53,120 --> 00:34:54,440 SMCHD 1 AND KNOCK IN LINE WHERE 887 00:34:54,440 --> 00:34:57,440 THERE IS LOSS OF DUX 4 AND 888 00:34:57,440 --> 00:35:00,600 RECOVERY OF SMCHD 1 AND HERE YOU 889 00:35:00,600 --> 00:35:02,840 CAN SEE RECOVERY OF CELLULAR 890 00:35:02,840 --> 00:35:05,760 VIABILITY IN THE KNOCK-IN LINE. 891 00:35:05,760 --> 00:35:08,640 BEALSO CONFIRMED DUX 4 892 00:35:08,640 --> 00:35:10,280 PATHOGENESIS, BY USING TWO 893 00:35:10,280 --> 00:35:13,000 DIFFERENT DUX 4 INHIBITORS. 894 00:35:13,000 --> 00:35:17,240 HERE WE USE SI RNA TO DIFFERENT 895 00:35:17,240 --> 00:35:19,960 SI RNAs TO KNOCK DOWN DUX 4. 896 00:35:19,960 --> 00:35:21,840 YOU CAN SEE WITH ONE OF THESE 897 00:35:21,840 --> 00:35:24,040 SHOWN HERE AGAIN, RECOVERY OF 898 00:35:24,040 --> 00:35:27,480 CELL VIABILITY. 899 00:35:27,480 --> 00:35:29,600 WE ALSO USE AD PHARMACOLOGICAL 900 00:35:29,600 --> 00:35:33,600 INHIBITOR CALLED IP 300W AND DUX 901 00:35:33,600 --> 00:35:37,440 4 ACTIVATES TARGET GENES RECRUIT 902 00:35:37,440 --> 00:35:40,320 RECRUITING THESE CO-ACTIVATORS 903 00:35:40,320 --> 00:35:42,040 AND CRED BINDING PROTEIN. 904 00:35:42,040 --> 00:35:43,920 WHEN YOU INHIBIT THESE PROTEINS 905 00:35:43,920 --> 00:35:47,240 WITH THE DRUG, WE SAW LOSS OF 906 00:35:47,240 --> 00:35:49,800 DUX 4 EXPRESSION, AND AGAIN 907 00:35:49,800 --> 00:35:54,200 TREATMENT WITH INHUB TORR 908 00:35:54,200 --> 00:35:55,440 RESCUES CELLS FROM CELL DEATH. 909 00:35:55,440 --> 00:35:56,840 WHAT ABOUT THE PATIENT DERIVED 910 00:35:56,840 --> 00:35:59,040 CELLS THAT I TOLD YOU ABOUT? 911 00:35:59,040 --> 00:36:00,400 WELL, THESE CELLS ACTUALLY 912 00:36:00,400 --> 00:36:02,480 BEHAVED QUITE SIMPLY TO THE 913 00:36:02,480 --> 00:36:03,320 KNOCK OUT CELLS. 914 00:36:03,320 --> 00:36:07,440 SO THIS IS SHOWING THE WILD TYPE 915 00:36:07,440 --> 00:36:09,440 CLONES AND TWO KNOCK OUT CLONES 916 00:36:09,440 --> 00:36:13,480 SHOWING CELL DEATH AT DAY 11 OF 917 00:36:13,480 --> 00:36:14,760 PLACODE CELL DIFFERENTIATION. 918 00:36:14,760 --> 00:36:18,080 HERE ARE IPS CELLS THAT WERE 919 00:36:18,080 --> 00:36:21,320 DIFFERENTIATED INTO PL,ACODE 920 00:36:21,320 --> 00:36:22,560 CELLS FROM HEALTHY PATIENT, 921 00:36:22,560 --> 00:36:25,760 THREE WITH ARHINIA, AND TWO 922 00:36:25,760 --> 00:36:27,240 PATIENTS WITH FSHD 2. 923 00:36:27,240 --> 00:36:29,120 AND YOU CAN SEE THAT THIS -- THE 924 00:36:29,120 --> 00:36:31,880 AMOUNT OF CELL DEATH CORRELATES 925 00:36:31,880 --> 00:36:33,800 WITH EXPRESSION OF THAT DUX 4 926 00:36:33,800 --> 00:36:36,640 TARGET I MENTIONED, TRIM 43 AS 927 00:36:36,640 --> 00:36:37,600 SHOPE HERE. 928 00:36:37,600 --> 00:36:40,120 AND I THINK WHAT IS INTERESTING 929 00:36:40,120 --> 00:36:44,280 IS THAT THE SAME VARIABILITY IN 930 00:36:44,280 --> 00:36:46,000 THESE CELLULAR PHENOTYPES THAT 931 00:36:46,000 --> 00:36:47,760 THIS IS VERY REMINISCENT OF THE 932 00:36:47,760 --> 00:36:50,440 VARIABILITY THAT WE SAW IN OUR 933 00:36:50,440 --> 00:36:52,240 PATIENTS AS WELL AND THIS AGAIN 934 00:36:52,240 --> 00:36:54,440 POINTS TO A ROLE FOR GENETIC OR 935 00:36:54,440 --> 00:36:56,600 ENVIRONMENTAL MODIFIERS, AND 936 00:36:56,600 --> 00:36:58,560 TELL US THAT THOSE MODIFIERS MAY 937 00:36:58,560 --> 00:37:00,080 ACTUALLY BE CONTROLLING THE 938 00:37:00,080 --> 00:37:03,200 AMOUNT OF DUX 4 EXPRESSION IN 939 00:37:03,200 --> 00:37:03,800 PLACODE CELLS. 940 00:37:03,800 --> 00:37:05,120 SO HOW ARE WE GOING TO FIND 941 00:37:05,120 --> 00:37:06,640 THESE? 942 00:37:06,640 --> 00:37:08,960 WELL, WE ARE PROPOSING TO 943 00:37:08,960 --> 00:37:12,040 PERFORM A GENOME WIDE SI RNA 944 00:37:12,040 --> 00:37:12,600 SCREEN. 945 00:37:12,600 --> 00:37:15,640 SO WE CREATED A CONSTRUCT WHERE 946 00:37:15,640 --> 00:37:18,720 DUX 4 BINDING TO ITS RESPONSIVE 947 00:37:18,720 --> 00:37:21,400 ELEMENT DRIVES EXPRESSION OF 948 00:37:21,400 --> 00:37:22,040 NANOLUCIFERASE. 949 00:37:22,040 --> 00:37:23,320 AND AS A NEGATIVE CONTROL WE 950 00:37:23,320 --> 00:37:26,440 HAVE JUST A PROMOTER LINKED TO 951 00:37:26,440 --> 00:37:27,760 NANOLUCIFERASE. 952 00:37:27,760 --> 00:37:30,880 AND WE TRANSFECTED OUR PLACODE 953 00:37:30,880 --> 00:37:32,880 CELLS AT DAY THREE OF 954 00:37:32,880 --> 00:37:35,360 DIFFERENTIATION, AND HERE YOU 955 00:37:35,360 --> 00:37:38,880 CAN SEE KNOWN NANOLUCIFERASE, 956 00:37:38,880 --> 00:37:40,320 STAYED NULL IN NEGATIVE CONTROL 957 00:37:40,320 --> 00:37:42,840 BUT IN THE KNOCK OUT CELLS WHERE 958 00:37:42,840 --> 00:37:45,680 WE EXPECTS TO SEE INCREASE DUX 4 959 00:37:45,680 --> 00:37:47,720 PRODUCTION AND BINDING WE SEE AN 960 00:37:47,720 --> 00:37:48,960 INCREASE IN NANOLUCIFERASE 961 00:37:48,960 --> 00:37:50,080 ACTIVITY. 962 00:37:50,080 --> 00:37:52,640 SO IN PERFORMING THIS SCREEN 963 00:37:52,640 --> 00:37:55,840 WHAT WE ARE LOOKING FOR IS GENES 964 00:37:55,840 --> 00:37:57,120 WHICH WHEN KNOCKED DOWN LEAD TO 965 00:37:57,120 --> 00:37:58,560 DECREASE IN SIGNAL IN OUR KNOCK 966 00:37:58,560 --> 00:38:00,960 OUT CELLS OR INCREASE IN SIGNAL 967 00:38:00,960 --> 00:38:06,440 IN WILD TYPE CELLS TO DO THIS 968 00:38:06,440 --> 00:38:07,640 FOR EVERY GENE IN THE GENOME WE 969 00:38:07,640 --> 00:38:09,560 COLLABORATE WITH NCATS WITH HIGH 970 00:38:09,560 --> 00:38:13,920 THROUGH PUT PLATE READER TO 971 00:38:13,920 --> 00:38:15,240 DETECT LUMINESCENCE. 972 00:38:15,240 --> 00:38:16,760 WHILE THIS IS COOKING WE HAVE 973 00:38:16,760 --> 00:38:20,040 BEGUN TO THINK ABOUT POTENTIAL 974 00:38:20,040 --> 00:38:21,600 ENVIRONMENTAL -- CAUSING 975 00:38:21,600 --> 00:38:22,360 ARHINIA. 976 00:38:22,360 --> 00:38:24,120 AND ONE SUCH CANDIDATE IS THE 977 00:38:24,120 --> 00:38:25,440 HERPES VIRUS FAMILY. 978 00:38:25,440 --> 00:38:28,600 THIS IS A RECENT PAPER BY FULL 979 00:38:28,600 --> 00:38:32,440 ET ALL IN NATURE MICROBIOLOGY 980 00:38:32,440 --> 00:38:34,440 DEMONSTRATING THAT TRIM 43 IS 981 00:38:34,440 --> 00:38:36,440 UPREGULATED DURING HERPES VIRUS 982 00:38:36,440 --> 00:38:38,960 INFECTION AS PART OF A GERM LINE 983 00:38:38,960 --> 00:38:41,320 SPECIFIC TRANSCRIPTIONAL PROGRAM 984 00:38:41,320 --> 00:38:43,120 MEDIATED BY NONE OTHER THAN DUX 985 00:38:43,120 --> 00:38:44,040 4. 986 00:38:44,040 --> 00:38:47,200 SO WE ASKED WHETHER HERPES VIRUS 987 00:38:47,200 --> 00:38:49,840 INFECTION OF OUR PATIENT DERIVED 988 00:38:49,840 --> 00:38:52,600 CELLS MIGHT SIMILARLY AUGMENT 989 00:38:52,600 --> 00:38:53,160 DUX 4. 990 00:38:53,160 --> 00:38:55,840 T THAT IS EXACTLY WHAT WE SAW. 991 00:38:55,840 --> 00:38:59,240 ON THE LEFT IS SHOWING CRE -- 992 00:38:59,240 --> 00:39:00,360 PREINFECTION AND ON THE RIGHT 993 00:39:00,360 --> 00:39:05,640 POST INFECTION WITH HSV 1, YOU 994 00:39:05,640 --> 00:39:07,480 CAN SEE IN THE PRESENCE OF VIRAL 995 00:39:07,480 --> 00:39:10,960 PROTEIN ICP 0 AND INDUCTION OF 996 00:39:10,960 --> 00:39:13,640 DUX 4 NOT IN CONTROL IPS CELLS, 997 00:39:13,640 --> 00:39:17,080 BUT IN CELLS FROM PATIENTS WITH 998 00:39:17,080 --> 00:39:18,240 FSHD 2 AND ARHINIA. 999 00:39:18,240 --> 00:39:21,160 HERE LOOKING AT TRIM 43 WE SEE 1000 00:39:21,160 --> 00:39:24,520 THE SAME PATTERN, SO NO INCREASE 1001 00:39:24,520 --> 00:39:27,240 PRE-INFECTION, AND A GREATER 1002 00:39:27,240 --> 00:39:32,240 INCREASE IN TRIM 43 AFTER HSV 1 1003 00:39:32,240 --> 00:39:34,240 INFECTION IN FSHD 2 AND PATIENTS 1004 00:39:34,240 --> 00:39:35,360 WITH ARHINIA CELLS. 1005 00:39:35,360 --> 00:39:37,440 LET'S PUT THIS ALL TOGETHER NOW. 1006 00:39:37,440 --> 00:39:39,960 WE STARTED WITH JUST ONE 1007 00:39:39,960 --> 00:39:40,640 PATIENT. 1008 00:39:40,640 --> 00:39:41,880 FORMED INTERNATIONAL CONSORTIUM 1009 00:39:41,880 --> 00:39:43,640 TO CREATE A LARGE COHORT OF 1010 00:39:43,640 --> 00:39:44,840 PATIENTS. 1011 00:39:44,840 --> 00:39:46,640 AND THROUGH NEXT GENERATION 1012 00:39:46,640 --> 00:39:48,680 SEQUENCING, WE WERE ABLE TO 1013 00:39:48,680 --> 00:39:50,360 DEMONSTRATE THAT MISSENSE 1014 00:39:50,360 --> 00:39:52,360 MUTATIONS IN SMCHD 1 ARE THE 1015 00:39:52,360 --> 00:39:54,640 PRIMARY DRIVER OF ARHINIA. 1016 00:39:54,640 --> 00:39:56,840 HOWEVER, THE HETEROGENEITY OF 1017 00:39:56,840 --> 00:39:58,840 PHENOTYPES THAT WE SAW IN OUR 1018 00:39:58,840 --> 00:40:00,480 PATIENTS, SUGGESTED THAT THERE 1019 00:40:00,480 --> 00:40:02,720 MAYBE ADDITIONAL GENETIC OR 1020 00:40:02,720 --> 00:40:05,720 ENVIRONMENTAL MODIFIERS. 1021 00:40:05,720 --> 00:40:09,160 WORK IN THE WET LAB DEMONSTRATED 1022 00:40:09,160 --> 00:40:12,040 THAT ARHINIA IS CAUSED BY DUX 4 1023 00:40:12,040 --> 00:40:14,600 MEDIATED PLACODE CELL DEATH AND 1024 00:40:14,600 --> 00:40:18,160 SUGGESTED A POTENTIAL ROLE FOR 1025 00:40:18,160 --> 00:40:21,600 HSV 1 IN PATHOGENESIS. 1026 00:40:21,600 --> 00:40:22,920 SO THERE ARE STILL SEVERAL OPEN 1027 00:40:22,920 --> 00:40:24,880 QUESTIONS THAT WE ARE TRYING TO 1028 00:40:24,880 --> 00:40:25,440 ADDRESS. 1029 00:40:25,440 --> 00:40:27,480 THE FIRST IS WHAT IS SPECIAL 1030 00:40:27,480 --> 00:40:30,040 ABOUT MUSCLE AND PLACODE CELLS 1031 00:40:30,040 --> 00:40:32,720 THAT ALLOWS FOR DUX 4 1032 00:40:32,720 --> 00:40:34,400 DEREPRESSION WITH LOSS OF SMCHD 1033 00:40:34,400 --> 00:40:34,760 1 ACTIVITY? 1034 00:40:34,760 --> 00:40:36,240 WHY IS IT LIMITED TO THESE CELL 1035 00:40:36,240 --> 00:40:36,640 TYPES? 1036 00:40:36,640 --> 00:40:38,520 AND WHY ARE THERE NO INDIVIDUALS 1037 00:40:38,520 --> 00:40:40,680 WITH BOTH DISORDERS? 1038 00:40:40,680 --> 00:40:44,520 DOES ONE PROTECT YOU FROM 1039 00:40:44,520 --> 00:40:46,560 GETTING THE OTHER DISORDER OR IS 1040 00:40:46,560 --> 00:40:49,880 IT THAT THE GENETIC CONDITIONS 1041 00:40:49,880 --> 00:40:51,320 ARE SO RARE THAT ONE INDIVIDUAL 1042 00:40:51,320 --> 00:40:54,000 IS UNLIKELY TO EVERY MEET 1043 00:40:54,000 --> 00:40:55,480 CRITERIA FOR BOTH CONDITIONS? 1044 00:40:55,480 --> 00:40:57,480 AND LASTLY, ARE THERE OTHER 1045 00:40:57,480 --> 00:40:58,600 ENVIRONMENTAL CONTRIBUTORS WE 1046 00:40:58,600 --> 00:41:01,240 SHOULD BE CONSIDERING? 1047 00:41:01,240 --> 00:41:03,160 I WANT TO THANK AND ACKNOWLEDGE 1048 00:41:03,160 --> 00:41:06,120 THE MANY COLLABORATORS WHO 1049 00:41:06,120 --> 00:41:07,680 HELPED THESE ARE ALL AUTHORS ON 1050 00:41:07,680 --> 00:41:10,360 OUR FIRST PAPER, IDENTIFYING 1051 00:41:10,360 --> 00:41:12,480 MUTATIONS IN SMCHD 1 AS CAUSE OF 1052 00:41:12,480 --> 00:41:13,280 ARHINIA. 1053 00:41:13,280 --> 00:41:15,960 AND THE MORE RECENT WORK 1054 00:41:15,960 --> 00:41:18,840 INVOLVING THE PHENOTYPING OF 1055 00:41:18,840 --> 00:41:22,000 PATIENTS AND THE PLACODE CELL 1056 00:41:22,000 --> 00:41:23,480 WORK PERFORMED BY THESE 1057 00:41:23,480 --> 00:41:25,160 INDIVIDUALS, I WANT TO THANK MY 1058 00:41:25,160 --> 00:41:28,240 COLLEAGUES AT THE NIH CLINICAL 1059 00:41:28,240 --> 00:41:31,560 CENTER AND AT NIEHS IN THE X-RAY 1060 00:41:31,560 --> 00:41:33,320 CRYSTALOGRAPHY FOR THE CLINICAL 1061 00:41:33,320 --> 00:41:34,040 RESEARCH UNIT AND 1062 00:41:34,040 --> 00:41:35,760 BIOINFORMATICS. 1063 00:41:35,760 --> 00:41:39,800 AND I WANT TO ESPECIALLY 1064 00:41:39,800 --> 00:41:45,360 HIGHLIGHT KAORU,NOUE WHO DID THE 1065 00:41:45,360 --> 00:41:49,280 PLA CODE AND STEM CELL WORK AND 1066 00:41:49,280 --> 00:41:52,560 COLLABORATORS AT MASS GENERAL. 1067 00:41:52,560 --> 00:41:55,040 THANK YOU FOR YOUR ATTENTION. 1068 00:41:55,040 --> 00:41:57,080 >> DR. SHAW, I WANT TO THANK YOU 1069 00:41:57,080 --> 00:42:00,680 FOR SHARING YOUR COMPLEX 1070 00:42:00,680 --> 00:42:02,720 INVESTIGATION INTO THIS UNUSUAL 1071 00:42:02,720 --> 00:42:06,920 PHENOTYPE AND INSIGHTS INTO THE 1072 00:42:06,920 --> 00:42:08,440 EPIGENETICS AS A FAST GAINING 1073 00:42:08,440 --> 00:42:11,840 PRESENTATION AND WE APPRECIATE 1074 00:42:11,840 --> 00:42:13,280 YOUR TIME. 1075 00:42:13,280 --> 00:42:15,640 I DO APOLOGIZE TO YOU AND THE 1076 00:42:15,640 --> 00:42:18,080 AUDIENCE FOR THE AUDIO BUT 1077 00:42:18,080 --> 00:42:19,040 THANKS TORR YOUR PATIENCE. 1078 00:42:19,040 --> 00:42:20,240 WE HAVE ONE QUESTION, LIKE TO 1079 00:42:20,240 --> 00:42:24,800 SHARE IT WITH YOU, SAYS DR. 1080 00:42:24,800 --> 00:42:27,480 SHAW, FANTASTIC PRESENTATION. 1081 00:42:27,480 --> 00:42:32,040 IF ARHINIA AND FSHD 2 ARE 1082 00:42:32,040 --> 00:42:33,680 MULTI-GENIC COMPLEX TRAITS, 1083 00:42:33,680 --> 00:42:37,800 WONDER IF STUDYING GENOME WIDE 1084 00:42:37,800 --> 00:42:39,040 PROFILES AND MODEL CELLS CAN BE 1085 00:42:39,040 --> 00:42:41,920 HELPFUL IN IDENTIFYING TADs 1086 00:42:41,920 --> 00:42:43,400 THAT ARE DIFFERENT FROM 1087 00:42:43,400 --> 00:42:46,760 FILTERING STRATEGY TO STUDY 1088 00:42:46,760 --> 00:42:48,440 EPISTATIC MACHINES. 1089 00:42:48,440 --> 00:42:50,840 >> THAT IS AN EXCELLENT POINT 1090 00:42:50,840 --> 00:42:52,240 AND I DON'T HAVE THE DATA TO 1091 00:42:52,240 --> 00:42:55,600 SHARE WITH YOU BUT WE HAVE 1092 00:42:55,600 --> 00:42:57,440 PERFORMED HIGH C STUDIES MANY 1093 00:42:57,440 --> 00:43:00,240 THE CRANIAL PLACODE CELLS AND WE 1094 00:43:00,240 --> 00:43:01,960 ARE ANALYZING THOSE DATA NOW BUT 1095 00:43:01,960 --> 00:43:03,960 THAT IS A GREAT SUGGESTION. 1096 00:43:03,960 --> 00:43:06,840 >> GREAT. 1097 00:43:06,840 --> 00:43:09,240 AND JUST ANY OTHER INSIGHTS INTO 1098 00:43:09,240 --> 00:43:11,240 -- YOU WERE LOOKING AT THE 1099 00:43:11,240 --> 00:43:13,320 ENVIRONMENTAL INFLUENCES AND 1100 00:43:13,320 --> 00:43:14,200 FOUND HERPES. 1101 00:43:14,200 --> 00:43:15,560 ARE THERE OTHER DIRECTIONS THAT 1102 00:43:15,560 --> 00:43:16,840 YOU ARE CURRENTLY LOOKING INTO 1103 00:43:16,840 --> 00:43:19,720 SPECIFICALLY RIGHT NOW? 1104 00:43:19,720 --> 00:43:20,520 >> YEAH. 1105 00:43:20,520 --> 00:43:22,320 WE DON'T -- WHEN I TALKED TO 1106 00:43:22,320 --> 00:43:25,640 THESE PATIENTS AND THEIR PARENTS 1107 00:43:25,640 --> 00:43:28,440 THERE'S REALLY NO -- THEY DON'T 1108 00:43:28,440 --> 00:43:31,240 MENTION ANY POTENTIAL EXPOSURES. 1109 00:43:31,240 --> 00:43:33,680 THEY HAVE BEEN TOLD DIFFERENT 1110 00:43:33,680 --> 00:43:35,040 THINGS THAT WERE CERTAINLY WRONG 1111 00:43:35,040 --> 00:43:37,360 LIKE MOTHER WAS TOO CLOSE TO THE 1112 00:43:37,360 --> 00:43:39,400 X-RAY -- COPY MACHINE AT HER 1113 00:43:39,400 --> 00:43:42,720 WORK OR THINGS LIKE THAT. 1114 00:43:42,720 --> 00:43:44,240 WHICH I JUST FEEL HORRIBLE WHEN 1115 00:43:44,240 --> 00:43:48,520 I HEAR -- BUT I THINK PROBABLY 1116 00:43:48,520 --> 00:43:52,000 WILL LOOK AT MORE VIRUS, BUT NO, 1117 00:43:52,000 --> 00:43:55,640 I DON'T HAVE ANY OTHER LEADS ON 1118 00:43:55,640 --> 00:43:56,400 POTENTIAL ENVIRONMENTAL 1119 00:43:56,400 --> 00:43:59,200 CONTRIBUTORS. 1120 00:43:59,200 --> 00:44:04,880 >> THANK YOU VERY MUCH. 1121 00:44:04,880 --> 00:44:06,840 LOOKS LIKE YOU HAVE ANSWERED ALL 1122 00:44:06,840 --> 00:44:08,600 THE QUESTIONS FROM THE AUDIENCE, 1123 00:44:08,600 --> 00:44:11,160 NOTHING ELSE IS COMING IN. 1124 00:44:11,160 --> 00:44:13,240 AGAIN, I WANT TO THANK YOU, VERY 1125 00:44:13,240 --> 00:44:14,560 MUCH FOR PRESENTING TODAY. 1126 00:44:14,560 --> 00:44:16,360 WE REALLY ENJOYED YOUR 1127 00:44:16,360 --> 00:44:17,120 PRESENTATION. 1128 00:44:17,120 --> 00:44:18,720 AND I WANT TO THANK -- WE HAVE 1129 00:44:18,720 --> 00:44:19,440 ONE MORE QUESTION JUST POPPED 1130 00:44:19,440 --> 00:44:24,440 IN. 1131 00:44:24,440 --> 00:44:27,720 IT IS A -- DR. SHAW, IS IT ALSO 1132 00:44:27,720 --> 00:44:29,160 POSSIBLE THESE TWO PHENOTYPES 1133 00:44:29,160 --> 00:44:30,320 HAVE NOT BEEN SEEN TOGETHER 1134 00:44:30,320 --> 00:44:34,080 BECAUSE OF SOME OTHER LINK THAT 1135 00:44:34,080 --> 00:44:36,320 LEAD TO LEE HALLTY IN UTERO? 1136 00:44:36,320 --> 00:44:38,160 -- LETHALITY IN UTERO? 1137 00:44:38,160 --> 00:44:39,920 >> I GUESS THAT'S POSSIBLE. 1138 00:44:39,920 --> 00:44:45,840 SO THAT WOULD -- YEAH, I MEAN, 1139 00:44:45,840 --> 00:44:48,040 SO YOU ARE SUGGESTING THAT IF 1140 00:44:48,040 --> 00:44:50,640 THERE WERE MORE SEVERE MUTATION, 1141 00:44:50,640 --> 00:44:54,440 THAT COULD CAUSE BOTH 1142 00:44:54,440 --> 00:44:55,640 CONDITIONS, IT WOULDN'T -- THE 1143 00:44:55,640 --> 00:44:59,440 CHILD WOULD NOT SURVIVE. 1144 00:44:59,440 --> 00:45:01,440 YEAH, I HAVEN'T THOUGHT OF THAT 1145 00:45:01,440 --> 00:45:04,960 BUT THAT ALSO COULD BE POSSIBLE. 1146 00:45:04,960 --> 00:45:09,200 THERE ARE NO -- IF YOU LOOK IN 1147 00:45:09,200 --> 00:45:13,040 NOMAD, THERE'S NO PATHOGEN I CAN 1148 00:45:13,040 --> 00:45:13,560 NULL MUTATION. 1149 00:45:13,560 --> 00:45:16,120 I SHOULDN'T SAY THAT, THAT CAUSE 1150 00:45:16,120 --> 00:45:18,000 ARHINIA, THEY ARE ALL MISSENSE. 1151 00:45:18,000 --> 00:45:21,360 BUT THERE ARE NO MUTATIONS THAT 1152 00:45:21,360 --> 00:45:24,360 CAUSE FSHD 2, THERE IS A SEVERE 1153 00:45:24,360 --> 00:45:27,960 FORM OF FSHD 2 THAT HAS A 1154 00:45:27,960 --> 00:45:30,800 NEONATAL ONSET THAT CAN CAUSE 1155 00:45:30,800 --> 00:45:34,880 MORE MUSCLE BUT ALSO 1156 00:45:34,880 --> 00:45:36,120 DEVELOPMENTAL DELAY BUT AGAIN, 1157 00:45:36,120 --> 00:45:39,640 THOSE PATIENTS HAVE NO CRANIAL 1158 00:45:39,640 --> 00:45:42,720 FACIAL ABNORMALITIES. 1159 00:45:42,720 --> 00:45:44,240 >> OKAY. 1160 00:45:44,240 --> 00:45:46,120 THANK YOU, VERY MUCH, DR. SHAW, 1161 00:45:46,120 --> 00:45:48,360 THANKS TO THE AUDIENCE. 1162 00:45:48,360 --> 00:45:49,440 I WISH EVERYONE A GREAT 1163 00:45:49,440 --> 00:45:50,440 AFTERNOON. 1164 00:45:50,440 --> 00:45:52,040 THANK YOU, VERY MUCH. 1165 00:45:52,040 --> 00:49:22,400 >> THANK YOU.