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 --> 00:00:47,560 Learn more by visiting us online at http://clinicalcenter.nih.gov 11 00:00:47,560 --> 00:00:49,960 TODAY WE WELCOME OUR SPEAKERS, 12 00:00:49,960 --> 00:00:51,520 THE CO-DIRECTORS OF THE 13 00:00:51,520 --> 00:00:52,680 TRANSLATIONAL PULMONARY ARTERIAL 14 00:00:52,680 --> 00:00:54,680 HYPERTENSION RESEARCH SECTION OF 15 00:00:54,680 --> 00:00:56,240 THE NIH CRITICAL CARE MEDICINE 16 00:00:56,240 --> 00:00:58,760 DEPARTMENT, DR. MICHAEL SOLOMON, 17 00:00:58,760 --> 00:01:00,480 SENIOR RESEARCH PHYSICIAN AND 18 00:01:00,480 --> 00:01:02,800 DR. JASON ELINOFF, TENURE TRACK 19 00:01:02,800 --> 00:01:03,800 INVESTIGATOR. 20 00:01:03,800 --> 00:01:05,960 OUR FIRST SPEAKER IS DR. 21 00:01:05,960 --> 00:01:07,400 SOLOMON, SENIOR RESEARCH 22 00:01:07,400 --> 00:01:08,480 PHYSICIAN, HEAD OF THE 23 00:01:08,480 --> 00:01:09,680 CARDIOLOGY SECTION OF CRITICAL 24 00:01:09,680 --> 00:01:11,440 CARE MEDICINE AND CO-DIRECTOR OF 25 00:01:11,440 --> 00:01:13,600 THE TRANSLATIONAL PULMONARY 26 00:01:13,600 --> 00:01:14,320 ARTERIAL HYPERTENSION RESEARCH 27 00:01:14,320 --> 00:01:17,400 SECTION. DR. SOLOMON ALSO HOLD 28 00:01:17,400 --> 00:01:20,080 AS JOINT APPOINTMENT IN THE 29 00:01:20,080 --> 00:01:21,280 CARDIOVASCULAR BRANCH NATIONAL 30 00:01:21,280 --> 00:01:23,840 HEART LUNG AND BLOOD INSTITUTE. 31 00:01:23,840 --> 00:01:25,600 DR. SOLOMON RECEIVED HIS 32 00:01:25,600 --> 00:01:26,840 UNDERGRADUATE DEGREE FROM BROWN 33 00:01:26,840 --> 00:01:28,720 UNIVERSITY AND EARNED HIS 34 00:01:28,720 --> 00:01:30,520 MEDICAL DEGREE FROM NEW YORK 35 00:01:30,520 --> 00:01:31,400 UNIVERSITY SCHOOL OF MEDICINE. 36 00:01:31,400 --> 00:01:32,960 HE COMPLETED INTERNAL MEDICINE 37 00:01:32,960 --> 00:01:33,960 INTERNSHIP AND RESIDENCY 38 00:01:33,960 --> 00:01:35,800 UNIVERSITY OF TEXAS SOUTH 39 00:01:35,800 --> 00:01:37,760 WESTERN MEDICAL CENTER. 40 00:01:37,760 --> 00:01:39,640 FOLLOWING RESIDENCY HE COMPLETED 41 00:01:39,640 --> 00:01:40,520 CRITICAL CARE MEDICINE 42 00:01:40,520 --> 00:01:42,560 FELLOWSHIP AT THE NIH. 43 00:01:42,560 --> 00:01:43,720 AFTERWARDS DR. SOLOMON RETURNED 44 00:01:43,720 --> 00:01:45,640 TO UT SOUTH WESTERN TO COMPLETE 45 00:01:45,640 --> 00:01:47,280 A SECOND FELLOWSHIP IN 46 00:01:47,280 --> 00:01:49,520 CARDIOVASCULAR DISEASES. 47 00:01:49,520 --> 00:01:53,040 HE JOINED A CARDIOLOGY FACULTY 48 00:01:53,040 --> 00:01:55,000 AT UT SOUTH WESTERN SPECIALIZING 49 00:01:55,000 --> 00:01:56,600 IN PATIENTS WITH PULMONARY 50 00:01:56,600 --> 00:01:57,640 HYPERTENSION, CARDIAC 51 00:01:57,640 --> 00:01:59,120 TRANSPLANTATION. 52 00:01:59,120 --> 00:02:01,120 HE RETURNED TO THE NIH AS HEAD 53 00:02:01,120 --> 00:02:02,560 OF THE CARDIOLOGY SECTION OF 54 00:02:02,560 --> 00:02:05,240 CRITICAL CARE MEDICINE IN 2000. 55 00:02:05,240 --> 00:02:07,480 MOST RECENTLY DR. SOLOMON EARNED 56 00:02:07,480 --> 00:02:10,760 HIS MBA IN 2000 WITH ADDITIONAL 57 00:02:10,760 --> 00:02:12,040 CERTIFICATE IN BUSINESS 58 00:02:12,040 --> 00:02:13,240 ANALYTICS FROM THE MASSACHUSETTS 59 00:02:13,240 --> 00:02:14,720 INSTITUTE OF TECHNOLOGY SCHROENN 60 00:02:14,720 --> 00:02:17,840 SCHOOL OF MANAGEMENT. 61 00:02:17,840 --> 00:02:19,480 DR. SOLOMON SUPERVISES 62 00:02:19,480 --> 00:02:20,640 ACTIVITIES RELATED TO THE FIELDS 63 00:02:20,640 --> 00:02:22,480 OF CRITICAL CARE CARDIOLOGY AND 64 00:02:22,480 --> 00:02:24,080 PULMONARY HYPERTENSION. 65 00:02:24,080 --> 00:02:26,520 HIS RESEARCH INTERESTS INCLUDE 66 00:02:26,520 --> 00:02:28,280 ENDOTHELIAL CELL DYSFUNCTION, 67 00:02:28,280 --> 00:02:30,280 INFLAMMATION IN PULMONARY 68 00:02:30,280 --> 00:02:32,320 ARTERIAL HYPERTENSION OR PAH. 69 00:02:32,320 --> 00:02:33,520 AS WELL AS ORGANIZATIONAL 70 00:02:33,520 --> 00:02:35,720 STRUCTURES AND OUTCOMES DELIVERY 71 00:02:35,720 --> 00:02:38,840 OF CRITICAL CARE CARDIOLOGY. 72 00:02:38,840 --> 00:02:40,640 DR. SOLOMON IS BOARD CERTIFIED 73 00:02:40,640 --> 00:02:42,520 IN INTERNAL MEDICINE, 74 00:02:42,520 --> 00:02:43,280 CARDIOVASCULAR DISEASE AND 75 00:02:43,280 --> 00:02:44,960 CRITICAL CARE MEDICINE BY THE 76 00:02:44,960 --> 00:02:45,880 AMERICAN BOARD OF INTERNAL 77 00:02:45,880 --> 00:02:47,200 MEDICINE. 78 00:02:47,200 --> 00:02:50,160 HE SERVES ON THE ADVISORY FOR 79 00:02:50,160 --> 00:02:51,920 THE NIH MEDICAL RESEARCH 80 00:02:51,920 --> 00:02:53,800 SCHOLARS PROGRAM, IS VICE CHAIR 81 00:02:53,800 --> 00:02:55,400 OF THE NIH STAFF CLINICIAN 82 00:02:55,400 --> 00:02:57,840 COUNCIL, AND IS CLAIRE OF THE 83 00:02:57,840 --> 00:02:59,280 NIH INARM SHY AND THERAPEUTICS 84 00:02:59,280 --> 00:02:59,920 COMMITTEE. 85 00:02:59,920 --> 00:03:01,760 -- PHARMACY AND THERAPEUTICS 86 00:03:01,760 --> 00:03:02,040 COMMITTEE. 87 00:03:02,040 --> 00:03:05,680 HE IS CO-CHAIR OF AMERICAN CARE 88 00:03:05,680 --> 00:03:07,360 CARDIOLOGY, LEADERSHIP COUNCIL. 89 00:03:07,360 --> 00:03:09,040 A SECOND SPEAKER TODAY IS DR. 90 00:03:09,040 --> 00:03:10,880 JASON ELINOFF. 91 00:03:10,880 --> 00:03:13,520 TENURE TRACK INVESTIGATOR, HEAD 92 00:03:13,520 --> 00:03:15,960 OF PULMONARY VASCULAR SECTION 93 00:03:15,960 --> 00:03:19,440 AND CO-DIRECTOR TRANSLATIONAL 94 00:03:19,440 --> 00:03:20,200 PULMONARY HYPERTENSION PROGRAM. 95 00:03:20,200 --> 00:03:21,880 DR. ELINOFF EARNED UNDERGRADUATE 96 00:03:21,880 --> 00:03:23,880 IN CHEMISTRY FROM PRINCETON 97 00:03:23,880 --> 00:03:25,200 UNIVERSITY, MEDICAL DEGREE FROM 98 00:03:25,200 --> 00:03:27,360 UNIVERSITY OF CHICAGO PRITCHARD 99 00:03:27,360 --> 00:03:29,080 SCHOOL OF MEDICINE. 100 00:03:29,080 --> 00:03:31,360 HE COMPLETED INTERNSHIP AND 101 00:03:31,360 --> 00:03:33,440 RESIDENCY TRAINING AT 102 00:03:33,440 --> 00:03:34,320 MASSACHUSETTS GENERAL HOSPITAL. 103 00:03:34,320 --> 00:03:36,040 DR. ELINOFF COMPLETED HIS 104 00:03:36,040 --> 00:03:37,920 FELLOWSHIP TRAINING CRITICAL 105 00:03:37,920 --> 00:03:38,920 CARE MEDICINE AT THE NIH 106 00:03:38,920 --> 00:03:40,400 CLINICAL CENTER AND PULMONARY 107 00:03:40,400 --> 00:03:42,080 AND CRITICAL CARE MEDICINE AT 108 00:03:42,080 --> 00:03:44,400 JOHNS HOPKINS UNIVERSITY. 109 00:03:44,400 --> 00:03:45,440 HEATHEN JOINED NIH CLINICAL 110 00:03:45,440 --> 00:03:48,560 CENTER AS STAFF CLINICIAN IN 111 00:03:48,560 --> 00:03:49,160 YEAR 2011. 112 00:03:49,160 --> 00:03:52,360 DR. ELINOFF RESEARCH FOCUS IS 113 00:03:52,360 --> 00:03:53,160 CONTRIBUTIONS OF PULMONARY 114 00:03:53,160 --> 00:03:56,400 VASCULAR ENDOTHELIUM AND IMMUNE 115 00:03:56,400 --> 00:03:57,400 EFFECTOR CELLS TO DEVELOP 116 00:03:57,400 --> 00:03:58,520 PROGRESSION OF PAH. 117 00:03:58,520 --> 00:04:00,600 HIS CURRENT WORK INCLUDES THE IN 118 00:04:00,600 --> 00:04:03,520 VITRO PROFILING OF HUMAN 119 00:04:03,520 --> 00:04:06,200 PULMONARY ARTERIAL ENDOTHELIAL 120 00:04:06,200 --> 00:04:07,880 CELLS WITH PH ASSOCIATED 121 00:04:07,880 --> 00:04:09,720 MOLECULAR DEFECTS TO INVESTIGATE 122 00:04:09,720 --> 00:04:11,200 MOLECULAR MECHANISM EXPLOITED 123 00:04:11,200 --> 00:04:13,400 THERAPEUTICALLY. 124 00:04:13,400 --> 00:04:14,320 THE OVERARCHING GOAL OF THE 125 00:04:14,320 --> 00:04:15,880 PROGRAM IS TRANSLATE PRE-SCIENCE 126 00:04:15,880 --> 00:04:17,960 AND PRE-CLINICAL STUDIES TO 127 00:04:17,960 --> 00:04:19,200 EARLY PHASE THERAPEUTIC TRIALS 128 00:04:19,200 --> 00:04:21,560 IN PAH PATIENTS. 129 00:04:21,560 --> 00:04:23,560 DR. ELINOFF IS BOARD CERTIFIED 130 00:04:23,560 --> 00:04:25,160 IN INTERNAL MEDICINE, PULMONARY 131 00:04:25,160 --> 00:04:26,360 MEDICINE AND CRITICAL CARE 132 00:04:26,360 --> 00:04:27,080 MEDICINE. 133 00:04:27,080 --> 00:04:29,600 THE TITLE OF OUR SPEAKERS 134 00:04:29,600 --> 00:04:32,520 PRESENTATION TODAY IS PULMONARY 135 00:04:32,520 --> 00:04:34,760 ARTERIAL HYPERTENSION, WHAT LIES 136 00:04:34,760 --> 00:04:36,840 BENEATH THE TIP OF THE ICEBERG. 137 00:04:36,840 --> 00:04:38,000 WELCOME FIRST SPEAKER DR. 138 00:04:38,000 --> 00:04:38,240 MICHAEL SOLOMON. 139 00:04:38,240 --> 00:04:39,920 >> GOOD AFTERNOON, EVERYONE. 140 00:04:39,920 --> 00:04:42,680 DR. ELINOFF AND I APPRECIATE THE 141 00:04:42,680 --> 00:04:44,400 OPPORTUNITY TO PRESENT TO OUR 142 00:04:44,400 --> 00:04:45,960 COLLEAGUES OUR EFFORTS TO 143 00:04:45,960 --> 00:04:47,880 UNDERSTAND AND BETTER TREAT THE 144 00:04:47,880 --> 00:04:49,760 RARE DISEASE PULMONARY ARTERIAL 145 00:04:49,760 --> 00:04:50,080 HYPERTENSION. 146 00:04:50,080 --> 00:04:52,560 TODAY WE WILL SHARE WITH YOU 147 00:04:52,560 --> 00:04:55,200 WHAT LIES BENEATH THE TIP OF THE 148 00:04:55,200 --> 00:04:55,800 ICEBERG. 149 00:04:55,800 --> 00:04:57,600 NEITHER DR. ELINOFF NOR I HAVE 150 00:04:57,600 --> 00:04:58,840 RELEVANT FINANCIAL DISCLOSURES 151 00:04:58,840 --> 00:05:01,000 RELATED TO THIS PRESENTATION. 152 00:05:01,000 --> 00:05:02,800 HOWEVER BOTH OF US WILL BE 153 00:05:02,800 --> 00:05:06,680 DISCUSSING THE OFF LABEL USE OF 154 00:05:06,680 --> 00:05:07,920 SPUR LACTONE IN PULL NO MARE 155 00:05:07,920 --> 00:05:09,640 HYPERTENSION. 156 00:05:09,640 --> 00:05:10,760 -- PULMONARY HYPERTENSION. 157 00:05:10,760 --> 00:05:12,080 OUR LEARNING OBJECTIVES BY THE 158 00:05:12,080 --> 00:05:13,280 CONCLUSION OF THE PRESENTATION 159 00:05:13,280 --> 00:05:14,480 YOU SHOULD BE ABLE TO DESCRIBE 160 00:05:14,480 --> 00:05:17,720 THE PATHOPHYSIOLOGY OF PULMONARY 161 00:05:17,720 --> 00:05:19,040 ARTERIAL HYPERTENSION, DESCRIBE 162 00:05:19,040 --> 00:05:20,920 HOW CELLULAR MODELS OF PAH 163 00:05:20,920 --> 00:05:22,600 ASSOCIATED MOLECULAR DEFECTS, 164 00:05:22,600 --> 00:05:25,240 CAN BE USED TO IDENTIFYING NEW 165 00:05:25,240 --> 00:05:27,160 THERAPEUTIC TARGETS. 166 00:05:27,160 --> 00:05:29,400 DISCUSS INFLAMMATION IN 167 00:05:29,400 --> 00:05:31,720 PULMONARY ARTERIAL HYPERTENSION, 168 00:05:31,720 --> 00:05:34,400 AND RECOGNIZE PROTEIN SPECIFIC 169 00:05:34,400 --> 00:05:37,040 DEGRADATION AS A STRATEGY FOR 170 00:05:37,040 --> 00:05:40,040 MODIFYING INFLAMMATION. 171 00:05:40,040 --> 00:05:42,040 PULMONARY ARTERIAL HYPERTENSION 172 00:05:42,040 --> 00:05:45,440 IS A RARE FEMALE PREDOMINANT 173 00:05:45,440 --> 00:05:45,920 DISEASE. 174 00:05:45,920 --> 00:05:48,280 PITTS WORLD HEALTH ORGANIZATION 175 00:05:48,280 --> 00:05:50,720 CLASSIFICATION IS GROUP 1. 176 00:05:50,720 --> 00:05:53,920 ITS PREVALENCE IS ESTIMATED AT 177 00:05:53,920 --> 00:05:57,160 15 TO 50 CASES PER MILLION. 178 00:05:57,160 --> 00:06:00,560 ITS INCIDENCE IS 2.4 CASES PER 179 00:06:00,560 --> 00:06:02,800 MILLION PER YEAR. 180 00:06:02,800 --> 00:06:04,360 PREDOMINANTLY EFFECTS WOMEN WHO 181 00:06:04,360 --> 00:06:07,000 CONSTITUTE 70 TO 80% OF 182 00:06:07,000 --> 00:06:08,440 PATIENTS, AGE AT DIAGNOSIS 183 00:06:08,440 --> 00:06:10,680 RANGES FROM 30 TO 60 YEARS. 184 00:06:10,680 --> 00:06:13,320 WITH THOSE HAVING THE IT OWE 185 00:06:13,320 --> 00:06:14,440 PATHIC VARIETY BEING YOUNGER AT 186 00:06:14,440 --> 00:06:16,280 DIAGNOSIS. 187 00:06:16,280 --> 00:06:20,160 THE MAJORITY OF WHO GROUP 1 188 00:06:20,160 --> 00:06:21,280 PULMONARY HYPERTENSION IS MADE 189 00:06:21,280 --> 00:06:24,120 UP OF IDIOPATHIC UNEXPLAINED 190 00:06:24,120 --> 00:06:27,480 VARIETY AND DISEASE ASSOCIATED. 191 00:06:27,480 --> 00:06:29,440 TOGETHER THEY ACCOUNT FOR 90% OF 192 00:06:29,440 --> 00:06:30,560 CASES. 193 00:06:30,560 --> 00:06:34,640 DISEASE ASSOCIATED PAH CONSISTS 194 00:06:34,640 --> 00:06:35,440 PREDOMINANTLY OF CONNECTIVE 195 00:06:35,440 --> 00:06:37,400 TISSUE DISEASE AND CONGENITAL 196 00:06:37,400 --> 00:06:39,880 HEART DISEASE, MAKING UP 80% OF 197 00:06:39,880 --> 00:06:42,440 THIS CATEGORY. 198 00:06:42,440 --> 00:06:46,440 THE MAJORITY OF WHO THERAPIES 199 00:06:46,440 --> 00:06:47,600 ARE SHOWN HERE ON THIS TIME 200 00:06:47,600 --> 00:06:48,560 LINE. 201 00:06:48,560 --> 00:06:50,880 THE TIME LINE ILLUSTRATES THE 202 00:06:50,880 --> 00:06:52,560 RELATIVELY REMARKABLE PACE OF 203 00:06:52,560 --> 00:06:54,320 PAH DRUG DEVELOPMENT, WE HAVE 204 00:06:54,320 --> 00:06:59,480 GONE FROM 0 TO 14 OPTIONS FOR 205 00:06:59,480 --> 00:07:01,760 TREATMENT OVER A 20 YEAR PERIOD. 206 00:07:01,760 --> 00:07:04,960 THEY TARGET THE ABNORMAL 207 00:07:04,960 --> 00:07:06,320 VASOCONSTRICTION OF THE 208 00:07:06,320 --> 00:07:07,640 DYSFUNCTIONAL ENDOTHELIUM; 209 00:07:07,640 --> 00:07:09,320 CURRENT THERAPIES WORK THROUGH 210 00:07:09,320 --> 00:07:12,400 THE ENDOTHREE LYNN, NITRIC OXIDE 211 00:07:12,400 --> 00:07:13,000 AND FROST CYCLINE PATHWAYS. 212 00:07:13,000 --> 00:07:15,080 ACTIVATION OF THE ENDOTHELIN 213 00:07:15,080 --> 00:07:17,960 PATHWAY RESULTS IN 214 00:07:17,960 --> 00:07:18,840 VASOCONSTRICTION THROUGH THE ET 215 00:07:18,840 --> 00:07:21,160 1 RECEPTOR. 216 00:07:21,160 --> 00:07:22,480 ENDOTHREE LYNN RECEPTOR 217 00:07:22,480 --> 00:07:24,240 ANTAGONIST BLOCK THIS EFFECT. 218 00:07:24,240 --> 00:07:27,520 THIS CLASS OF ORAL THERAPIES 219 00:07:27,520 --> 00:07:31,480 CONSIST OF AMBERSENTIN, THE 220 00:07:31,480 --> 00:07:34,240 SENTINEL STUDY WAS THE BOSENTIN 221 00:07:34,240 --> 00:07:36,880 (PHONETIC) RANDOMIZED TRIAL OF 222 00:07:36,880 --> 00:07:39,080 ENDOTHREE LYNN ANTAGONIST 223 00:07:39,080 --> 00:07:40,680 THERAPY, ENDOTHELIN THERAPY. 224 00:07:40,680 --> 00:07:42,440 THE PRIMARY END POINT WAS A 225 00:07:42,440 --> 00:07:43,600 CHANGE IN SIX MINUTE WALK 226 00:07:43,600 --> 00:07:45,320 DISTANCE WITH A SIGNIFICANT 227 00:07:45,320 --> 00:07:48,160 INCREASE WITH BOSENTIN COMPARED 228 00:07:48,160 --> 00:07:48,640 TO PLACEBO. 229 00:07:48,640 --> 00:07:51,400 THE NITRIC OXIDE PATHWAY ACTS 230 00:07:51,400 --> 00:07:55,400 THROUGH CYCLIC GMP TO CAUSE 231 00:07:55,400 --> 00:07:57,280 VASODILATION. 232 00:07:57,280 --> 00:07:59,920 PHOSPHODIE ESTERASE FIVE 233 00:07:59,920 --> 00:08:02,280 METABOLIZES CYCLIC GMP 234 00:08:02,280 --> 00:08:03,320 INHIBITING PATHWAY. 235 00:08:03,320 --> 00:08:06,840 TO RESTORE VASODILATION WE TREAT 236 00:08:06,840 --> 00:08:09,680 WITH PHOSPHODEESTERASE 237 00:08:09,680 --> 00:08:12,760 INHIBITORS OR A CYCLASE 238 00:08:12,760 --> 00:08:14,160 STIMULATOR. 239 00:08:14,160 --> 00:08:17,600 PD 5 INHIBITORS AND REOSIGWA 240 00:08:17,600 --> 00:08:21,080 (PHONETIC) IS A CYCLIC GMP 241 00:08:21,080 --> 00:08:21,600 STIMULATOR. 242 00:08:21,600 --> 00:08:25,240 THE SENTINEL STUDY WAS SEDINOFIL 243 00:08:25,240 --> 00:08:27,320 USE IN PAH, THE SUPER ONE TRIAL, 244 00:08:27,320 --> 00:08:30,120 THE PRIMARY END POINT WAS AGAIN 245 00:08:30,120 --> 00:08:30,880 CHANGING SIX MINUTE WALK 246 00:08:30,880 --> 00:08:32,800 DISTANCE WITH A SIGNIFICANT 247 00:08:32,800 --> 00:08:35,480 INCREASE WITH SELDINOFIL 248 00:08:35,480 --> 00:08:38,360 COMPARED TO PLACEBO. 249 00:08:38,360 --> 00:08:40,560 NOTABLY 20, 40 AND 80 MILLIGRAMS 250 00:08:40,560 --> 00:08:41,240 WERE NOT DIFFERENT FROM EACH 251 00:08:41,240 --> 00:08:44,360 OTHER. 252 00:08:44,360 --> 00:08:47,440 THE FROST CYCLINE PATHWAY ACTS 253 00:08:47,440 --> 00:08:49,000 THROUGH OPPRESS CYCLIC PATHWAY 254 00:08:49,000 --> 00:08:51,760 ENDS TO DERIVATIVES AND RECEPTOR 255 00:08:51,760 --> 00:08:55,040 AGONIST STIMULATE CYCLIC AMP, 256 00:08:55,040 --> 00:08:55,640 THE FROST CYCLINE DERIVATIVE 257 00:08:55,640 --> 00:08:59,640 COMES IN ORAL, INHALED, SUB Q 258 00:08:59,640 --> 00:09:00,640 AND INTRAVENOUS FORMS. 259 00:09:00,640 --> 00:09:02,640 THE LATTER TWO BEING GIVEN 260 00:09:02,640 --> 00:09:06,440 CONTINUOUSLY THROUGH A PUMP. 261 00:09:06,440 --> 00:09:08,640 ILOPROST (PHONETIC) IS INHALED 262 00:09:08,640 --> 00:09:10,640 AND IT IS GIVEN INTRAVENOUSLY AS 263 00:09:10,640 --> 00:09:11,320 CONTINUES WAS INFUSION. 264 00:09:11,320 --> 00:09:15,480 THE HALF LIFE OF IPOPROSNOL IS 265 00:09:15,480 --> 00:09:18,520 ORDER OF MINUTES WHILE 266 00:09:18,520 --> 00:09:19,840 TREPROSNOL IS FOUR AND A HALF 267 00:09:19,840 --> 00:09:20,840 HOURS. 268 00:09:20,840 --> 00:09:23,360 THE ORRILL AGENT IS THE ONLY FDA 269 00:09:23,360 --> 00:09:26,280 APPROVED ORAL AGENT IS THE ONLY 270 00:09:26,280 --> 00:09:29,640 FDA PROS STOW CYCLINE AGONIST. 271 00:09:29,640 --> 00:09:32,920 EPO WAS STUDIED IN HIGHLY 272 00:09:32,920 --> 00:09:34,160 SYMPTOMATIC PATIENTS NEW YORK 273 00:09:34,160 --> 00:09:36,280 HEART ASSOCIATION CLASS 3, 4, 274 00:09:36,280 --> 00:09:37,160 PATIENTS WITH MARK ACTIVITY 275 00:09:37,160 --> 00:09:38,320 LIMITATIONS. 276 00:09:38,320 --> 00:09:40,960 IT IS THE ONLY MEDICATION THAT 277 00:09:40,960 --> 00:09:42,640 IS MET END POINT OF IMPROVED 278 00:09:42,640 --> 00:09:46,200 SURVIVAL IN THE PAH POPULATION. 279 00:09:46,200 --> 00:09:48,160 NOW LET'S BRIEFLY DISCUSS OUR 280 00:09:48,160 --> 00:09:50,760 USAGE OF THESE MEDICATIONS. 281 00:09:50,760 --> 00:09:52,640 FOR PATIENTS WHO ARE NEW YORK 282 00:09:52,640 --> 00:09:55,040 HEART ASSOCIATION FUNCTIONAL 283 00:09:55,040 --> 00:09:57,280 CLASS 2, THOSE WHO ORDINARY 284 00:09:57,280 --> 00:09:59,920 PHYSICAL ACTIVITY CAUSES 285 00:09:59,920 --> 00:10:01,600 FATIGUE, WE RECOMMEND DUAL 286 00:10:01,600 --> 00:10:03,800 THERAPY WITH AN ERA AND A PD 5 287 00:10:03,800 --> 00:10:05,360 INHIBITOR. 288 00:10:05,360 --> 00:10:07,880 THIS IS IN PART BASED ON THE 289 00:10:07,880 --> 00:10:09,680 AMBITION TRIAL WHICH COMPARED 290 00:10:09,680 --> 00:10:13,960 DUAL THERAPY WITH AMBERSENTIN TO 291 00:10:13,960 --> 00:10:15,760 MONOTHERAPY WITH EITHER DRUG. 292 00:10:15,760 --> 00:10:18,440 AND FOUND COMBINATION THERAPY TO 293 00:10:18,440 --> 00:10:20,320 LOWER THE RISK OF CLINICAL 294 00:10:20,320 --> 00:10:20,920 WORSENING. 295 00:10:20,920 --> 00:10:23,440 THERE'S ALSO RECENT DATA 296 00:10:23,440 --> 00:10:25,240 SUGGESTING THAT INITIAL TRIPLE 297 00:10:25,240 --> 00:10:28,240 ORAL THERAPY REDUCES DISEASE 298 00:10:28,240 --> 00:10:29,880 PROGRESSION OVER INITIAL DOUBLE 299 00:10:29,880 --> 00:10:32,320 ORAL THERAPY, THOUGH THE HAZARD 300 00:10:32,320 --> 00:10:34,800 RATIO SLIGHTLY CROSSED OVER 301 00:10:34,800 --> 00:10:37,640 IDENTITIES SPANNING 0.23 TO 302 00:10:37,640 --> 00:10:38,040 1.079. 303 00:10:38,040 --> 00:10:40,520 HOWEVER WITH THIS IN MIND, 304 00:10:40,520 --> 00:10:42,640 PATIENTS WHO ARE INTERMEDIATE 305 00:10:42,640 --> 00:10:44,920 RISK SHOULD RECEIVE TRIPLE ORAL 306 00:10:44,920 --> 00:10:46,680 THERAPY WITH ORAL AGENTS ACTIVE 307 00:10:46,680 --> 00:10:49,880 ON ALL THREE PATHWAYS. 308 00:10:49,880 --> 00:10:53,720 ENDOTHELIN, NITRIC OXIDE AND 309 00:10:53,720 --> 00:10:54,400 FROST CYCLINE PATHWAYS. 310 00:10:54,400 --> 00:10:56,160 PATIENTS FAILING A PD 5 311 00:10:56,160 --> 00:10:59,160 INHIBITOR SHOULD BE CONSIDERED 312 00:10:59,160 --> 00:11:00,800 THE GAIN LACE CYCLASE STIMULATOR 313 00:11:00,800 --> 00:11:04,680 AS AN ALTERNATIVE AGENT. 314 00:11:04,680 --> 00:11:05,400 THOSE PATIENTS INTERMEDIATE TO 315 00:11:05,400 --> 00:11:07,960 HIGH RISK WITH DISMIA FATIGUE 316 00:11:07,960 --> 00:11:09,360 WITH LESS THAN ORDINARY ACTIVITY 317 00:11:09,360 --> 00:11:12,040 SHOULD BE CHANGED FROM ORAL 318 00:11:12,040 --> 00:11:12,800 FROST SIGH LYNN PATHWAY AGENTS 319 00:11:12,800 --> 00:11:18,280 TO INHALED OR CONTINUOUS IV OR 320 00:11:18,280 --> 00:11:21,600 SUB Q FROST CYCLINE DERIVATIVES. 321 00:11:21,600 --> 00:11:22,960 IF APPROPRIATE REFERRED TO 322 00:11:22,960 --> 00:11:25,720 TRANSPLANT CENTER FOR WORK UP. 323 00:11:25,720 --> 00:11:27,200 PATIENTS WHO NEW YORK HEART 324 00:11:27,200 --> 00:11:37,720 ASSOCIATION CLASS 4 HIGH RISK. 325 00:11:39,280 --> 00:11:41,200 THESE CANDIDATES SHOULD BE FOR 326 00:11:41,200 --> 00:11:44,280 CONTINUOUS EPOPROTNOL,HE ONLY 327 00:11:44,280 --> 00:11:45,560 MEDICATION SHOWN TO HAVE 328 00:11:45,560 --> 00:11:46,560 SURVIVAL BENEFIT IN NEW YORK 329 00:11:46,560 --> 00:11:48,680 HEART ASSOCIATION CLASS 4 PAH 330 00:11:48,680 --> 00:11:49,720 PATIENTS. 331 00:11:49,720 --> 00:11:51,000 THEY ALSO SHOULD BE REFERRED TO 332 00:11:51,000 --> 00:11:53,520 A TRANSPLANT CENTER FOR TIMELY 333 00:11:53,520 --> 00:11:55,040 WORK UP. 334 00:11:55,040 --> 00:11:57,760 THE MOST COMMON CAUSE OF DEATH 335 00:11:57,760 --> 00:11:59,320 IN PULMONARY ARTERIAL 336 00:11:59,320 --> 00:12:00,560 HYPERTENSION PATIENTS IS RIGHT 337 00:12:00,560 --> 00:12:03,600 HEART FAILURE. 338 00:12:03,600 --> 00:12:05,520 DEPICTED HERE ARE TWO ECHO 339 00:12:05,520 --> 00:12:07,160 CARDIOGRAPHIC VIEWS OF NORMAL 340 00:12:07,160 --> 00:12:09,080 LEFT AND RIGHT VENTRICLE. 341 00:12:09,080 --> 00:12:11,320 NOTE THE LEFT VENTRICLE THE 342 00:12:11,320 --> 00:12:14,000 LARGER THAN THE RIGHT AND IN 343 00:12:14,000 --> 00:12:15,520 CROSS SECTION BELOW THE LEFT 344 00:12:15,520 --> 00:12:17,640 VENTRICLE IS CIRCULAR IN SHAPE. 345 00:12:17,640 --> 00:12:20,280 IN PATIENTS WITH PULMONARY 346 00:12:20,280 --> 00:12:21,160 ARTERIAL HYPERTENSION, 347 00:12:21,160 --> 00:12:23,400 PROGRESSIVE NARROWING AND 348 00:12:23,400 --> 00:12:26,480 OBLITERATION OF THE DISTAL SMALL 349 00:12:26,480 --> 00:12:31,480 PULMONPULMONARY ARTERIES LEADS O 350 00:12:31,480 --> 00:12:33,080 INCREASES IN PULMONARY VASCULAR 351 00:12:33,080 --> 00:12:36,040 RESISTANCE, WORSENING OF RV 352 00:12:36,040 --> 00:12:38,560 FUNCTION, AND A DROP IN CARDIAC 353 00:12:38,560 --> 00:12:41,560 OUTPUT AND EVENTUALLY DEATH DUE 354 00:12:41,560 --> 00:12:43,320 TO RIGHT HEART FAILURE. 355 00:12:43,320 --> 00:12:45,800 SHOWN ON THE RIGHT ARE TWO ECHO 356 00:12:45,800 --> 00:12:47,600 CARDIOGRAPHIC VIEWS OF A PATIENT 357 00:12:47,600 --> 00:12:48,800 WITH ADVANCED PULMONARY ARTERIAL 358 00:12:48,800 --> 00:12:50,640 HYPERTENSION. 359 00:12:50,640 --> 00:12:53,640 IN STARK CONTRAST TO OUR HEALTHY 360 00:12:53,640 --> 00:12:55,880 PATIENT ECHOCARDIOGRAMS THESE 361 00:12:55,880 --> 00:12:57,320 REVEAL A HYPERTROPHIED AND 362 00:12:57,320 --> 00:12:59,480 MASSIVELY DILATED RIGHT 363 00:12:59,480 --> 00:13:00,920 VENTRICLE, THAT IS MUCH LARGER 364 00:13:00,920 --> 00:13:03,080 THAN THE LEFT VENTRICLE. 365 00:13:03,080 --> 00:13:04,640 THE RIGHT VENTRICLE COMPRESSES 366 00:13:04,640 --> 00:13:06,840 THE LEFT VENTRICLE INTO A D 367 00:13:06,840 --> 00:13:09,120 SHAPE WITH THE SEPTUM NOW 368 00:13:09,120 --> 00:13:11,160 BULGING TO THE LV AS SHOWN IN 369 00:13:11,160 --> 00:13:13,640 CROSS SECTION IN THE LOWER RIGHT 370 00:13:13,640 --> 00:13:14,920 IMAGE. 371 00:13:14,920 --> 00:13:17,680 THE RIGHT ATRIUM SIMILARLY 372 00:13:17,680 --> 00:13:19,360 COMPROMISES LEFT ATRIAL FILLING. 373 00:13:19,360 --> 00:13:21,400 THE I PAS DYSFUNCTION ALVING 374 00:13:21,400 --> 00:13:22,800 RIGHT ATRIUM AND RIGHT VENTRICLE 375 00:13:22,800 --> 00:13:25,240 HAVE COMPROMISED LV PRE-LOAD AND 376 00:13:25,240 --> 00:13:27,640 FUNCTION. 377 00:13:27,640 --> 00:13:30,280 THE PATIENT REQUIRED LIFE SAVING 378 00:13:30,280 --> 00:13:31,360 LUNG TRANSPLANT A FEW WEEKS 379 00:13:31,360 --> 00:13:33,280 AFTER THIS HE CAN COWAS 380 00:13:33,280 --> 00:13:35,720 PERFORMED AS -- ECHO WAS 381 00:13:35,720 --> 00:13:37,320 PERFORMED AS PARTS OF OUR 382 00:13:37,320 --> 00:13:39,240 NATURAL HISTORY STUDY. 383 00:13:39,240 --> 00:13:41,520 SURVIVOR DATA FROM THE 1980s 384 00:13:41,520 --> 00:13:43,360 PRIOR TO THE AVAILABILITY OF 385 00:13:43,360 --> 00:13:46,560 MOST CURRENT THERAPIES REVEALED 386 00:13:46,560 --> 00:13:48,400 50% MORTALITY AT THREE YEARS 387 00:13:48,400 --> 00:13:50,560 AFTER DIAGNOSTIC RIGHT HEART 388 00:13:50,560 --> 00:13:52,440 CATHETERIZATION. 389 00:13:52,440 --> 00:13:56,640 THE REVEAL REGISTERED SPANNING 390 00:13:56,640 --> 00:13:59,440 2001 TO 2009 CAPTURES THE IMPACT 391 00:13:59,440 --> 00:14:01,400 OF CURRENT THERAPIES ON 392 00:14:01,400 --> 00:14:02,880 SURVIVAL. 393 00:14:02,880 --> 00:14:04,760 UNFORTUNATELY MEDIAN SURVIVAL 394 00:14:04,760 --> 00:14:07,240 HAS BEEN EXTENDED BY ONLY A FEW 395 00:14:07,240 --> 00:14:10,680 YEARS WITH A 50% MORTALITY 396 00:14:10,680 --> 00:14:13,520 BETWEEN 5 TO 7 YEARS AFTER 397 00:14:13,520 --> 00:14:14,680 DIAGNOSTIC RIGHT HEART 398 00:14:14,680 --> 00:14:17,280 CATHETERIZATION. 399 00:14:17,280 --> 00:14:20,520 THUS, DESPITE 14 FDA APPROVED 400 00:14:20,520 --> 00:14:23,200 PULMONARY VASODILATED THERAPIES, 401 00:14:23,200 --> 00:14:25,680 MORTALITY REMAINS VERY HIGH. 402 00:14:25,680 --> 00:14:28,800 THERE MUST BE PATHOLOGIC 403 00:14:28,800 --> 00:14:30,040 PATHWAYS WE ARE NOT TREATING. 404 00:14:30,040 --> 00:14:31,960 IDENTIFYING THESE PATHWAYS WILL 405 00:14:31,960 --> 00:14:34,040 REQUIRE A DEEP DIVE BELOW THE 406 00:14:34,040 --> 00:14:35,280 TIM OF THE PULMONARY 407 00:14:35,280 --> 00:14:37,440 HYPERTENSION ICEBERG. 408 00:14:37,440 --> 00:14:39,960 IN ORDER TO FURTHER EXPLORE AND 409 00:14:39,960 --> 00:14:41,800 EXPLOIT OTHER TREATABLE 410 00:14:41,800 --> 00:14:43,320 PATHWAYS, WE MUST ACKNOWLEDGE 411 00:14:43,320 --> 00:14:45,440 SOME FOUNDATIONAL PRINCIPLES SO 412 00:14:45,440 --> 00:14:48,360 WE STUDY THE CORRECT PATIENT 413 00:14:48,360 --> 00:14:48,960 POPULATION. 414 00:14:48,960 --> 00:14:53,960 FIRST AMONG THEM, IS NOT ALL 415 00:14:53,960 --> 00:14:55,120 PULMONARY HYPERTENSION IS 416 00:14:55,120 --> 00:14:57,200 PULMONARY ARTERIAL HYPERTENSION. 417 00:14:57,200 --> 00:14:59,640 THE WORLD HEALTH ORGANIZATION 418 00:14:59,640 --> 00:15:01,400 CLINICAL CLASSIFICATION 419 00:15:01,400 --> 00:15:02,840 DISTINGUISHES FIVE SEPARATE 420 00:15:02,840 --> 00:15:05,480 GROUPS OF PULMONARY HYPERTENSION 421 00:15:05,480 --> 00:15:07,720 BASED ON CLINICAL 422 00:15:07,720 --> 00:15:10,600 CHARACTERISTICS, UNDERLYING 423 00:15:10,600 --> 00:15:11,400 PATHOBIOLOGY, AND DISTINCT 424 00:15:11,400 --> 00:15:12,360 TREATMENT APPROACHES. 425 00:15:12,360 --> 00:15:14,440 THESE ARE GROUP 1, OUR GROUP OF 426 00:15:14,440 --> 00:15:15,920 INTEREST, WHICH IS PULMONARY 427 00:15:15,920 --> 00:15:17,920 HYPERTENSION DUE TO PULMONARY 428 00:15:17,920 --> 00:15:20,320 VASCULAR DISEASE, GROUP 2 A MUCH 429 00:15:20,320 --> 00:15:22,000 LARGER GROUP WHICH IS PULMONARY 430 00:15:22,000 --> 00:15:24,760 HYPERTENSION DUE TO LEFT HEART 431 00:15:24,760 --> 00:15:26,760 DISEASE, GROUP 3 ALSO FAIRLY 432 00:15:26,760 --> 00:15:28,600 LARGE GROUP WHICH IS PULMONARY 433 00:15:28,600 --> 00:15:30,200 HYPERTENSION DUE TO PULMONARY 434 00:15:30,200 --> 00:15:32,400 PARYNCHEMAL DISEASE, AND 435 00:15:32,400 --> 00:15:34,320 HYPOXIA, GROUP 4 WHICH IS PH DUE 436 00:15:34,320 --> 00:15:36,960 TO CHRONIC THROMBO EMBOLIC 437 00:15:36,960 --> 00:15:38,960 PULMONARY DISEASE, AND GROUP 5, 438 00:15:38,960 --> 00:15:41,480 PH DUE TO UNCLEAR OR 439 00:15:41,480 --> 00:15:44,200 MULTI-FACTORIAL MECHANISMS. 440 00:15:44,200 --> 00:15:46,920 ABOVE THE SURFACE, ALL OF THESE 441 00:15:46,920 --> 00:15:48,400 GROUPS LOOK SIMILAR, WITH 442 00:15:48,400 --> 00:15:51,480 DYSPNEA AND FATIGUE, DUE TO 443 00:15:51,480 --> 00:15:52,800 ELEVATED PULMONARY ARTERY 444 00:15:52,800 --> 00:15:54,720 PRESSURE AND RV DYSFUNCTION. 445 00:15:54,720 --> 00:15:58,200 EVEN JUST BELOW THE SURFACE, 446 00:15:58,200 --> 00:16:00,160 HEMODYNAMICS CAN BE SIMILAR AS 447 00:16:00,160 --> 00:16:04,200 GROUPS 1, 3, 4 AND 5 ALL PRESENT 448 00:16:04,200 --> 00:16:07,600 AS PRE-CAPILLARY PH. 449 00:16:07,600 --> 00:16:10,720 THE 2006, SORRY THE 2018 SIX 450 00:16:10,720 --> 00:16:12,840 WORLD SYMPOSIUM DEFINE 451 00:16:12,840 --> 00:16:15,000 PRE-CAPILLARY PH AS A MEAN 452 00:16:15,000 --> 00:16:16,240 PULMONARY ARTERY PRESSURE 453 00:16:16,240 --> 00:16:19,280 GREATER THAN 20-MILLIMETERS OF 454 00:16:19,280 --> 00:16:21,040 MERCURY PLUS PULMONARY ARTERY 455 00:16:21,040 --> 00:16:22,160 OCCLUSION PRESSURE OF LESS THAN 456 00:16:22,160 --> 00:16:25,920 OR EQUAL TO 15-MILLIMETERS OF 457 00:16:25,920 --> 00:16:28,480 MERCURY AWE LONG WITH PULMONARY 458 00:16:28,480 --> 00:16:30,280 VASCULAR RESISTANCE OF THREE 459 00:16:30,280 --> 00:16:31,000 UNITS. 460 00:16:31,000 --> 00:16:32,640 ISOLATED POST CAPILLARY PH 461 00:16:32,640 --> 00:16:34,960 REPRESENTATIVE OF GROUP 2 HAS A 462 00:16:34,960 --> 00:16:37,560 SIMILAR MEAN PULMONARY ARTERY 463 00:16:37,560 --> 00:16:38,560 PRESSURE BUT THEIR PULMONARY 464 00:16:38,560 --> 00:16:40,000 ARTERY OCCLUSION PRESSURE IS 465 00:16:40,000 --> 00:16:42,240 GREATER THAN 15-MILLIMETER OF 466 00:16:42,240 --> 00:16:43,400 MERCURY AND THEIR PULMONARY 467 00:16:43,400 --> 00:16:46,080 VASCULAR RESISTANCE IS LESS THAN 468 00:16:46,080 --> 00:16:46,880 3 WOOD UNITS. 469 00:16:46,880 --> 00:16:49,080 HOWEVER IT GETS MORE 470 00:16:49,080 --> 00:16:49,560 COMPLICATED. 471 00:16:49,560 --> 00:16:52,360 GROUP 2 PH CAN PRESENT AS 472 00:16:52,360 --> 00:16:54,160 COMBINED PRE AND POST CAPILLARY 473 00:16:54,160 --> 00:16:55,200 PH. 474 00:16:55,200 --> 00:16:57,240 IN WHICH CASE THEIR PBR WOULD BE 475 00:16:57,240 --> 00:17:00,240 GREATER THAN OR EQUAL TO 3 WOOD 476 00:17:00,240 --> 00:17:01,320 UNITS AND SIMILAR TO 477 00:17:01,320 --> 00:17:04,040 PRE-CAPILLARY PH. 478 00:17:04,040 --> 00:17:07,200 SO RATHER THAN ONLY TREATING THE 479 00:17:07,200 --> 00:17:08,200 ELEVATED PULMONARY ARTERIAL 480 00:17:08,200 --> 00:17:09,360 PRESSURE IN THE RIGHT 481 00:17:09,360 --> 00:17:11,120 VENTRICULAR DISFUNCTION AT THE 482 00:17:11,120 --> 00:17:13,200 TIP OF THE ICEBERG AND 483 00:17:13,200 --> 00:17:15,200 PRE-CAPILLARY HEMODYNAMIC 484 00:17:15,200 --> 00:17:16,280 DERANGEMENTS IN SHALLOW WATERS 485 00:17:16,280 --> 00:17:17,520 WE LEAD TO LOOK DEEPER IN ORDER 486 00:17:17,520 --> 00:17:19,840 TO APPRECIATE THE COMPLEX 487 00:17:19,840 --> 00:17:21,920 INTERRELATED MOLECULAR PROCESSES 488 00:17:21,920 --> 00:17:24,160 SOME OF WHICH ARE ESSENTIALLY 489 00:17:24,160 --> 00:17:27,640 IGNORED, BY CURRENT TREATMENTS. 490 00:17:27,640 --> 00:17:28,560 ENDOTHELIUM DYSFUNCTION, THE 491 00:17:28,560 --> 00:17:30,920 FOCUS OF CURRENT PH THERAPIES, 492 00:17:30,920 --> 00:17:33,120 IS CHARACTERIZED BY ENDOTHELIAL 493 00:17:33,120 --> 00:17:35,920 TO MESENCHYMAL TRANSITION AND 494 00:17:35,920 --> 00:17:38,360 IMPAIRED VASOREACTIVITY. 495 00:17:38,360 --> 00:17:39,680 MESENCHYMAL TRANSITION IS A 496 00:17:39,680 --> 00:17:42,120 PROCESS WHEREBY AN ENDOTHELIAL 497 00:17:42,120 --> 00:17:44,360 CELL UNDERGOES A SERIES OF 498 00:17:44,360 --> 00:17:46,440 MOLECULAR EVENTS THAT LEADS TO 499 00:17:46,440 --> 00:17:48,560 CHANGE IN PHENOTYPE TOWARDS 500 00:17:48,560 --> 00:17:49,960 MESENCHYMAL TYPE CELL, 501 00:17:49,960 --> 00:17:50,800 FIBROBLASTS OR SMOOTH MUSCLE 502 00:17:50,800 --> 00:17:52,800 CELL. 503 00:17:52,800 --> 00:17:54,800 ABNORMAL CELLULAR PROLIFERATION 504 00:17:54,800 --> 00:17:58,880 IS CHARACTERIZED BY MYOGENESIS, 505 00:17:58,880 --> 00:18:01,240 ANGIOGENESIS AND VASCULAR SMOOTH 506 00:18:01,240 --> 00:18:03,120 MUSCLE CELLS THAT ARE 507 00:18:03,120 --> 00:18:05,120 HYPERPROLIFERATIVE AND RESISTANT 508 00:18:05,120 --> 00:18:07,800 TO APOPTOSIS. 509 00:18:07,800 --> 00:18:09,800 EMERGING PARADIGMS INCLUDE 510 00:18:09,800 --> 00:18:12,240 METABOLIC REPROGRAMMING, IN 511 00:18:12,240 --> 00:18:14,680 SIMILAR FASHION TO CANCER, 512 00:18:14,680 --> 00:18:16,160 PROLIFERATING CELLS AND 513 00:18:16,160 --> 00:18:16,840 PULMONARY ARTERIAL HYPERTENSION 514 00:18:16,840 --> 00:18:20,120 SWITCH THEIR METABOLISM TOWARDS 515 00:18:20,120 --> 00:18:22,360 AEROBIC GLYCOLYSIS, ASSOCIATED 516 00:18:22,360 --> 00:18:24,040 WITH VASCULAR EXPANSION. 517 00:18:24,040 --> 00:18:27,800 IN ADDITION THERE IS ALSO 518 00:18:27,800 --> 00:18:29,840 EVIDENCE OF MYO-- MITOCHONDRIAL 519 00:18:29,840 --> 00:18:32,480 DYSFUNCTION, RELATED TO 520 00:18:32,480 --> 00:18:36,120 BIOGENESIS AND BIOENERGETICS. 521 00:18:36,120 --> 00:18:37,520 INFLAMMATION IN PULMONARY 522 00:18:37,520 --> 00:18:38,960 ARTERIAL HYPERTENSION IS 523 00:18:38,960 --> 00:18:41,680 CHARACTERIZED BY PULMONARY 524 00:18:41,680 --> 00:18:43,360 PERIVASCULAR PRO INFLAMMATORY 525 00:18:43,360 --> 00:18:44,760 INFILTRATES AND CIRCULATING 526 00:18:44,760 --> 00:18:47,160 INFLAMMATORY CYTOKINES. 527 00:18:47,160 --> 00:18:49,480 THERE IS EVIDENCE OF BOTH LOCAL 528 00:18:49,480 --> 00:18:51,960 AND SYSTEMIC IMMUNE ACTIVATION 529 00:18:51,960 --> 00:18:52,400 IN PULMONARY ARTERIAL 530 00:18:52,400 --> 00:18:54,000 HYPERTENSION. 531 00:18:54,000 --> 00:18:57,040 IT IS ALSO NOTABLE THAT 532 00:18:57,040 --> 00:18:59,400 AUTOIMMUNE DISEASES INFECTION 533 00:18:59,400 --> 00:19:00,560 AND PRIOR INTERFERON TREATMENT 534 00:19:00,560 --> 00:19:02,680 ARE RISK FACTORS FOR PULMONARY 535 00:19:02,680 --> 00:19:05,440 ARTERIAL HYPERTENSION. 536 00:19:05,440 --> 00:19:07,080 NONE, I REPEAT, NONE OF THE 537 00:19:07,080 --> 00:19:09,240 CURRENT THERAPIES FOCUS ON 538 00:19:09,240 --> 00:19:11,040 INFLAMMATORY MEDIATORS OF 539 00:19:11,040 --> 00:19:14,320 VASCULAR REMODELING. 540 00:19:14,320 --> 00:19:16,400 SHOWN HERE IS A CROSS SECTION OF 541 00:19:16,400 --> 00:19:19,000 A NORMAL PULMONARY ARTERY. 542 00:19:19,000 --> 00:19:21,880 THIS IS AN ILLUSTRATION OF 543 00:19:21,880 --> 00:19:23,520 LUMINAL NARROWING DUE TO 544 00:19:23,520 --> 00:19:25,000 CELLULAR PROLIFERATION IN THE 545 00:19:25,000 --> 00:19:28,640 LIGHT RED CIRCLE, ALSO PRESENT 546 00:19:28,640 --> 00:19:30,520 IS PERIVASCULAR INFLAMMATION 547 00:19:30,520 --> 00:19:31,800 INSIDE THE PINK OUTLINE. 548 00:19:31,800 --> 00:19:35,040 IN ADDITION, THE EXTENT OF 549 00:19:35,040 --> 00:19:37,120 VESSEL REMODELING CORRELATES 550 00:19:37,120 --> 00:19:40,200 WITH THE SEVERITY OF THESE 551 00:19:40,200 --> 00:19:42,560 PERIVASCULAR INFLAMMATORY 552 00:19:42,560 --> 00:19:43,160 INFILTRATES. 553 00:19:43,160 --> 00:19:46,920 WE NOW REALIZE THAT IN PULMONARY 554 00:19:46,920 --> 00:19:48,080 ARTERIAL HYPERTENSION P THE 555 00:19:48,080 --> 00:19:51,080 PULMONARY VASCULAR LESIONS 556 00:19:51,080 --> 00:19:53,440 SUBSIST IN AN INFLAMMATORY 557 00:19:53,440 --> 00:19:55,960 MICROENVIRONMENT. 558 00:19:55,960 --> 00:19:57,720 SYSTEMIC EVIDENCE OF 559 00:19:57,720 --> 00:19:59,360 INFLAMMATION CAN BE FOUND FROM 560 00:19:59,360 --> 00:20:01,800 STUDIES COMPARING SERUM CYTOKINE 561 00:20:01,800 --> 00:20:04,240 LEVELS IN IDIOPATHIC PULMONARY 562 00:20:04,240 --> 00:20:05,600 ARTERIAL HYPERTENSION PATIENTS 563 00:20:05,600 --> 00:20:07,440 TO HEALTHY CONTROLS. 564 00:20:07,440 --> 00:20:11,400 IL 2, IL 6, IL 8 ARE 565 00:20:11,400 --> 00:20:14,960 INFLAMMATION RELATED CYTOKINES. 566 00:20:14,960 --> 00:20:18,040 IL 2, IL 6 AND IL 8 ARE ALL 567 00:20:18,040 --> 00:20:22,120 SIGNIFICANTLY ELEVATED IN 568 00:20:22,120 --> 00:20:23,440 IDIOPATHIC PULMONARY ARTERIAL 569 00:20:23,440 --> 00:20:24,560 HYPERTENSION PATIENTS COMPARED 570 00:20:24,560 --> 00:20:27,240 TO CONTROLS. 571 00:20:27,240 --> 00:20:29,280 KEY POINTS SO FAR. 572 00:20:29,280 --> 00:20:32,800 PAH IS CURRENTLY AN INCURABLE 573 00:20:32,800 --> 00:20:34,720 HIGH MORTALITY DISEASE DUE TO 574 00:20:34,720 --> 00:20:36,200 PULMONARY VASCULAR REMODELING 575 00:20:36,200 --> 00:20:38,040 LEADING TO VESSEL NARROWING AND 576 00:20:38,040 --> 00:20:40,400 RIGHT VENTRICULAR FAILURE. 577 00:20:40,400 --> 00:20:43,640 MAJOR DRIVERS OF UNDERLYING 578 00:20:43,640 --> 00:20:46,200 PATHOBIOLOGY ARE ENDOTHELIAL 579 00:20:46,200 --> 00:20:47,880 DYSFUNCTION, ABNORMAL CELLULAR 580 00:20:47,880 --> 00:20:49,760 PROLIFERATION, INFLAMMATION, AND 581 00:20:49,760 --> 00:20:51,800 METABOLIC REPROGRAMMING. 582 00:20:51,800 --> 00:20:56,000 CURRENT THERAPIES DO NOT TARGET 583 00:20:56,000 --> 00:20:56,600 INFLAMMATION. 584 00:20:56,600 --> 00:21:00,200 DR. ELINOFF WILL NOW REGALE YOU 585 00:21:00,200 --> 00:21:02,960 WITH HIS BENCH WORK DELINEATING 586 00:21:02,960 --> 00:21:06,240 INFLAMMATORY PATHWAYS IN PH. 587 00:21:06,240 --> 00:21:09,200 I WILL PASS THE BATON FOR 588 00:21:09,200 --> 00:21:11,840 SPEAKING TO DR. ELINOFF. 589 00:21:11,840 --> 00:21:12,160 >>THANK YOU, MICHAEL. 590 00:21:12,160 --> 00:21:14,040 AS HIGHLIGHTED BY DR. SOLOMON 591 00:21:14,040 --> 00:21:16,040 DESPITE THERAPEUTIC ADVANCES PAH 592 00:21:16,040 --> 00:21:18,720 REMAINS INCURABLE DISEASE. 593 00:21:18,720 --> 00:21:20,000 IMPORTANTLY, CURRENT THERAPIES 594 00:21:20,000 --> 00:21:21,560 DON'T TARGET INFLAMMATION 595 00:21:21,560 --> 00:21:24,800 UNDERLYING PAH PATHOBIOLOGY. 596 00:21:24,800 --> 00:21:25,720 THEREFORE OUR PROGRAMS GOAL IS 597 00:21:25,720 --> 00:21:28,400 TO IDENTIFY AND DEVELOP 598 00:21:28,400 --> 00:21:29,680 THERAPIES THAT TARGET VASCULAR 599 00:21:29,680 --> 00:21:30,480 INFLAMMATION AND VESSEL 600 00:21:30,480 --> 00:21:31,560 REMODELING. 601 00:21:31,560 --> 00:21:33,400 TODAY I'M GOING TO SHARE TWO 602 00:21:33,400 --> 00:21:34,520 STORIES OF HOW OUR WORK IN THE 603 00:21:34,520 --> 00:21:36,600 LAB IS LAYING THE FOUNDATION FOR 604 00:21:36,600 --> 00:21:38,280 TRANSLATION OF NEW TREATMENT 605 00:21:38,280 --> 00:21:39,800 APPROACHES INTO CLINICAL 606 00:21:39,800 --> 00:21:40,320 STUDIES. 607 00:21:40,320 --> 00:21:41,880 IN THE FIRST STORY I WILL 608 00:21:41,880 --> 00:21:44,800 DESCRIBE THE ANTI-INFLAMMATORY 609 00:21:44,800 --> 00:21:52,160 EFFECTS OF SCONOLACTONE DURING 610 00:21:52,160 --> 00:21:54,360 SCLERODERMA PI PIGMENTOSA, 611 00:21:54,360 --> 00:21:55,600 LEVERAGING CELLULAR MODELS OF 612 00:21:55,600 --> 00:21:57,920 PAH ASSOCIATED MOLECULAR DEFECTS 613 00:21:57,920 --> 00:22:02,120 TO IDENTIFY NEW THERAPEUTIC 614 00:22:02,120 --> 00:22:02,760 TARGETS. 615 00:22:02,760 --> 00:22:04,960 SPIRIT LACTONE IS A CORTICOID 616 00:22:04,960 --> 00:22:07,000 RECEPTOR ANTAGONIST THAT 617 00:22:07,000 --> 00:22:08,760 IMPROVES ENDSTHELIAL DISFUNCTION 618 00:22:08,760 --> 00:22:10,640 REDUCES I INFLAMMATION AND 619 00:22:10,640 --> 00:22:11,640 INCREASES SURVIVAL WITH LEFT 620 00:22:11,640 --> 00:22:12,560 HEART FAILURE. 621 00:22:12,560 --> 00:22:15,680 IT IS KNOWN TO HAVE BOTH MR 622 00:22:15,680 --> 00:22:17,400 DEPENDENT AND INDEPENDENT 623 00:22:17,400 --> 00:22:18,800 ANTI-INFLAMMATORY EFFECTS. 624 00:22:18,800 --> 00:22:21,240 HOWEVER, DESPITE DECADES OF 625 00:22:21,240 --> 00:22:22,920 RESEARCH, THE MOLECULAR 626 00:22:22,920 --> 00:22:24,400 MECHANISMS RESPONSIBLE FOR MR 627 00:22:24,400 --> 00:22:25,720 INDEPENDENT EFFECTS WERE POORLY 628 00:22:25,720 --> 00:22:28,200 UNDERSTAND. 629 00:22:28,200 --> 00:22:28,920 -- UNDERSTOOD. 630 00:22:28,920 --> 00:22:30,360 WE ESTABLISHED A MODEL SYSTEM IN 631 00:22:30,360 --> 00:22:32,520 HUMAN EMBRYONIC KIDNEY CELLS 632 00:22:32,520 --> 00:22:34,960 THAT DIDN'T EXPRESS STEROID HER 633 00:22:34,960 --> 00:22:36,480 HONE RECEPTORS THEREFORE ABLE TO 634 00:22:36,480 --> 00:22:37,720 SELECTIVELY EXPRESS THOSE 635 00:22:37,720 --> 00:22:40,480 RECEPTORS KNOWN TO INTERACT WITH 636 00:22:40,480 --> 00:22:41,400 SPURNO LACTONE. 637 00:22:41,400 --> 00:22:44,400 USING THE MODEL SYSTEM TWO 638 00:22:44,400 --> 00:22:46,080 CENTRAL INFLAMMATORY SIGNALING 639 00:22:46,080 --> 00:22:48,640 PATHWAYS NF KAPPA B AN AP 1 WERE 640 00:22:48,640 --> 00:22:49,960 INVESTIGATED USING APPROXIMATE 641 00:22:49,960 --> 00:22:53,080 LUCIFERASE REPORTER SYSTEM. 642 00:22:53,080 --> 00:22:57,680 HERE N NF KAPPA B IS ON Y AXIS 643 00:22:57,680 --> 00:22:58,960 RELATIVE TO IMIS THE RATION WITH 644 00:22:58,960 --> 00:23:01,840 TNF ALPHA, VALUES ALONG THE LINE 645 00:23:01,840 --> 00:23:02,680 SIGNIFY NO EFFECT. 646 00:23:02,680 --> 00:23:04,240 BELOW THE LINE SIGNIFIES 647 00:23:04,240 --> 00:23:07,160 SUPPRESSION OF TNF INDUCED 648 00:23:07,160 --> 00:23:07,760 ACTIVITY. 649 00:23:07,760 --> 00:23:11,720 INCREASING CONCENTRATIONS OF 650 00:23:11,720 --> 00:23:13,800 SPURNO LACTONE SHOWN ON THE 651 00:23:13,800 --> 00:23:14,640 RIGHT AXIS. 652 00:23:14,640 --> 00:23:16,520 ON THE RIGHT THEY ARE CONTROL 653 00:23:16,520 --> 00:23:18,960 VECTOR IN RED OR MR SPECIFIC 654 00:23:18,960 --> 00:23:20,200 RECEPTOR IN BLUE. 655 00:23:20,200 --> 00:23:21,480 DEMONSTRATING THE DIFFERENCES IN 656 00:23:21,480 --> 00:23:22,760 MR EXPRESSION. 657 00:23:22,760 --> 00:23:24,440 HERE YOU SEE THAT IN THE ABSENCE 658 00:23:24,440 --> 00:23:27,880 OF MR EXPRESSION, DOSE 659 00:23:27,880 --> 00:23:30,400 DEPENDENTLY SUPPRESSED NF KAPPA 660 00:23:30,400 --> 00:23:31,880 B ACTIVITY AN EFFECT NOT 661 00:23:31,880 --> 00:23:33,440 SIGNIFICANTLY ALTERED WHEN MR IS 662 00:23:33,440 --> 00:23:35,120 HIGHLY EXPRESSED. 663 00:23:35,120 --> 00:23:37,040 TO SUMMARIZE OUR FINDINGS AT 664 00:23:37,040 --> 00:23:38,720 THIS POINT INCLUDING DATA I DID 665 00:23:38,720 --> 00:23:40,360 NOT HAVE TIME TO SHOW, WE FOUND 666 00:23:40,360 --> 00:23:42,720 THAT SPURNO LACTONE SUPPRESSED 667 00:23:42,720 --> 00:23:45,760 NF KAPPA B AND AP 1 PROMOTER 668 00:23:45,760 --> 00:23:48,160 ACTIVITY INDEPENDENT OF THE 669 00:23:48,160 --> 00:23:49,280 MINERALOCORTICOID RECEPTOR. 670 00:23:49,280 --> 00:23:50,720 THESE EFFECTS WERE INDEPENDENT 671 00:23:50,720 --> 00:23:53,680 OF ANDROGEN, PROGESTERONE AND 672 00:23:53,680 --> 00:23:54,400 GLUCOCORTICOID RECEPTOR 673 00:23:54,400 --> 00:23:56,000 EXPRESSION. 674 00:23:56,000 --> 00:24:00,640 AND A MORE SELECTIVE ANTAGONIST 675 00:24:00,640 --> 00:24:01,880 DID NOT SUPPRESS INFLAMMATION IN 676 00:24:01,880 --> 00:24:02,360 THIS SYSTEM. 677 00:24:02,360 --> 00:24:05,040 A PREVIOUS STUDY IDENTIFIED 678 00:24:05,040 --> 00:24:07,520 SPURNO LACTONE OF INHIBITOR OF 679 00:24:07,520 --> 00:24:11,240 NUCLEOTIDE EXCISION REPAIR, A 680 00:24:11,240 --> 00:24:13,240 SPECIFIC DNA REPAIR MECHANISM, 681 00:24:13,240 --> 00:24:15,240 IN SETTING OF RADIATION, CELLS 682 00:24:15,240 --> 00:24:17,440 RECRUIT REPAIR MECHANISMS 683 00:24:17,440 --> 00:24:19,440 INCLUDING THE GENERAL 684 00:24:19,440 --> 00:24:22,720 TRANSCRIPTION FACTOR TH 2H. 685 00:24:22,720 --> 00:24:24,000 IN CELLS TREAT TREATED, DNA 686 00:24:24,000 --> 00:24:25,200 REPAIR WAS INHIBITED. 687 00:24:25,200 --> 00:24:27,680 TF 2H IS A PROTEIN COMPLEX 688 00:24:27,680 --> 00:24:31,440 COMPOSED OF 11 SUBUNITS 689 00:24:31,440 --> 00:24:33,880 INCLUDING IS SCLERODERMA PIG MEN 690 00:24:33,880 --> 00:24:39,880 TO IS A OR XPB. 691 00:24:39,880 --> 00:24:43,040 THEY HAVE A CENTRAL ROLE IN RNA 692 00:24:43,040 --> 00:24:44,640 TRANSCRIPTION AND DNA REPAIR. 693 00:24:44,640 --> 00:24:46,240 THEY DEMONSTRATED IT INHIBITED 694 00:24:46,240 --> 00:24:47,920 DNA REPAIR AS A RESULT OF 695 00:24:47,920 --> 00:24:49,440 PROTEOSOME DEPENDENT 696 00:24:49,440 --> 00:24:51,280 DEGRADATION. 697 00:24:51,280 --> 00:24:53,480 HOWEVER, THE LINK BETWEEN THIS 698 00:24:53,480 --> 00:24:55,600 EFFECT AND ITS ANTI-INFLAMMATORY 699 00:24:55,600 --> 00:24:57,080 PROPERTIES HAD NOT BEEN 700 00:24:57,080 --> 00:24:57,880 PREVIOUSLY EXPLORE. 701 00:24:57,880 --> 00:24:59,840 WE RETURNED TO OUR REPORTER 702 00:24:59,840 --> 00:25:04,400 SYSTEM TO INVESTIGATE WHETHER 703 00:25:04,400 --> 00:25:06,160 DEGRADATION CONTRIBUTES TO THE 704 00:25:06,160 --> 00:25:07,560 ANTI-INFLABTORY EFFECTS. 705 00:25:07,560 --> 00:25:10,200 IT SUPPRESSES AP 1 ACTIVITY 706 00:25:10,200 --> 00:25:12,040 INDUCED BY PMA, HOWEVER THIS 707 00:25:12,040 --> 00:25:13,160 EFFECT WAS REVERSED IN THE 708 00:25:13,160 --> 00:25:14,320 PRESENCE OF A PROTEOSOME 709 00:25:14,320 --> 00:25:15,920 INHIBITOR. 710 00:25:15,920 --> 00:25:17,480 UNDER THE SAME EXPERIMENTAL 711 00:25:17,480 --> 00:25:19,560 CONDITIONS AS OUR REPORTER 712 00:25:19,560 --> 00:25:22,760 ASSAY, WE FOUND SPIRN LACTONE 713 00:25:22,760 --> 00:25:27,280 TRIGGERS XPV DEGRADATION, 714 00:25:27,280 --> 00:25:29,200 REVERSED AS INHIBITOR BY WESTERN 715 00:25:29,200 --> 00:25:29,720 BLOT BELOW. 716 00:25:29,720 --> 00:25:31,400 WE OVEREXPRESSED IN OUR MODEL 717 00:25:31,400 --> 00:25:33,040 SYSTEM TO IN ORDER TO STRENGTHEN 718 00:25:33,040 --> 00:25:35,400 THE EVIDENCE THAT SPIRNO LACTONE 719 00:25:35,400 --> 00:25:37,200 SUPPRESSES INFLAMMATION AS A 720 00:25:37,200 --> 00:25:39,880 RESULT OF XPB LOSS. 721 00:25:39,880 --> 00:25:44,360 IT SUPPRESSED ACTIVITY AND 722 00:25:44,360 --> 00:25:46,560 REDUCED XPV LEVELS SHOWN BELOW. 723 00:25:46,560 --> 00:25:47,480 IN THE PRESENCE OF 724 00:25:47,480 --> 00:25:48,520 OVEREXPRESSION THE EFFECT IS 725 00:25:48,520 --> 00:25:51,360 COMPLETELY REVERSED. 726 00:25:51,360 --> 00:25:52,600 SIMILARLY ON THE RIGHT IT 727 00:25:52,600 --> 00:25:54,960 SUPPRESSED NF KAPPA B ACTIVITY 728 00:25:54,960 --> 00:25:57,400 AND REDUCED XPB LEVELS BUT AGAIN 729 00:25:57,400 --> 00:26:01,000 WITH OVEREXPRESSION OF XPV BOTH 730 00:26:01,000 --> 00:26:02,400 EFFECTS ARE REVERSED SO TO 731 00:26:02,400 --> 00:26:04,240 SUMMARIZE THE RESULTS OF THESE 732 00:26:04,240 --> 00:26:07,120 EXPERIMENTS, SPIRNO LACTONE 733 00:26:07,120 --> 00:26:09,320 TRIGGERS PROTEOSOME DEGRADATION 734 00:26:09,320 --> 00:26:11,080 OF XPV AND BLOCKING DEGRADATION 735 00:26:11,080 --> 00:26:13,520 WITH EITHER PROTEOSOME INHIBITOR 736 00:26:13,520 --> 00:26:18,160 OR OVEREXPRESSING XPB REVERSES 737 00:26:18,160 --> 00:26:19,960 SPURNO LACTONE MEDIATED 738 00:26:19,960 --> 00:26:22,000 SUPPRESSION AND NF KAPPA B 739 00:26:22,000 --> 00:26:22,600 PROMOTER ACTIVITY. 740 00:26:22,600 --> 00:26:24,840 WE TRANSLATED FINDINGS TO HUMAN 741 00:26:24,840 --> 00:26:26,320 PULMONARY ARTERY ENDOTHELIAL 742 00:26:26,320 --> 00:26:27,680 CELLS AND INVESTIGATED THE 743 00:26:27,680 --> 00:26:30,560 EFFECT OF SPIRNO LACTONE TNF 744 00:26:30,560 --> 00:26:32,960 INDUCED INFLAMMATORY MEDIATORS 745 00:26:32,960 --> 00:26:35,080 WITH PH PATHOGENESIS. 746 00:26:35,080 --> 00:26:36,880 IL 8 EXPRESSION IS SHOWN HERE 747 00:26:36,880 --> 00:26:38,840 PLOTTED ON THE Y AXIS, IN RED 748 00:26:38,840 --> 00:26:42,040 TNF INDUCES IL 8 TRANSCRIPTION 749 00:26:42,040 --> 00:26:44,960 SUPPRESS BY SPIRNO LACTONE. 750 00:26:44,960 --> 00:26:46,920 ON THE RIGHT CELL SUPERNATANT 751 00:26:46,920 --> 00:26:49,480 COLLECTED AT 4, 8 AND 24 HOURS 752 00:26:49,480 --> 00:26:52,240 ON THE X AXIS AND LOG 753 00:26:52,240 --> 00:26:52,800 TRANSFOREIGN PROTEIN 754 00:26:52,800 --> 00:26:53,920 CONCENTRATION ON THE Y AXIS. 755 00:26:53,920 --> 00:26:56,760 AS EXPECT WED SEE INCREASE IN IL 756 00:26:56,760 --> 00:26:59,040 8 SECRETION IN PRESENCE OF TNF 757 00:26:59,040 --> 00:27:00,640 ALPHA STIMULATION AND IT 758 00:27:00,640 --> 00:27:02,120 SUPPRESSES SECRETION AT EACH 759 00:27:02,120 --> 00:27:03,560 TIME POINT. 760 00:27:03,560 --> 00:27:05,480 IT ALSO SUPPRESSED A VARIETY OF 761 00:27:05,480 --> 00:27:07,560 OTHER TNF INDUCED INFLAMMATORY 762 00:27:07,560 --> 00:27:09,880 MEDIATORS INCLUDING CYTOKINES, 763 00:27:09,880 --> 00:27:10,920 CHEMOKINES, AND ADHESION 764 00:27:10,920 --> 00:27:12,600 MOLECULES. 765 00:27:12,600 --> 00:27:15,640 NEXT WE UTILIZE 766 00:27:15,640 --> 00:27:17,640 IMMUNOCYTOCHEMISTRY TO DETERMINE 767 00:27:17,640 --> 00:27:19,720 XPV PROTEIN EXPRESSION IN 768 00:27:19,720 --> 00:27:20,120 ENDOTHELIUM. 769 00:27:20,120 --> 00:27:21,840 CELLS ARE STIMULATE WITH TNF 770 00:27:21,840 --> 00:27:24,800 ALPHA ALONE ON THE LEFT, XPV 771 00:27:24,800 --> 00:27:26,240 STAINED IN GREEN AND ACTIN IN 772 00:27:26,240 --> 00:27:26,400 RED. 773 00:27:26,400 --> 00:27:27,440 ON THE RIGHT YOU CAN SEE IN 774 00:27:27,440 --> 00:27:29,760 CELLS TREATED WITH SPIRNO 775 00:27:29,760 --> 00:27:32,120 LACTONE THERE IS DECREASE IN XPV 776 00:27:32,120 --> 00:27:33,680 EXPRESSION. 777 00:27:33,680 --> 00:27:37,080 AS SHOWN EARLIER, IT 778 00:27:37,080 --> 00:27:40,160 SIGNIFICANTLY SUPPRESSED IL 8 779 00:27:40,160 --> 00:27:43,040 mRNA EXPRESSION IN PULMONARY 780 00:27:43,040 --> 00:27:44,200 ARTERY ENDOTHELIAL CELLS. 781 00:27:44,200 --> 00:27:46,560 WE IDENTIFIED I P KAPPA RNA 782 00:27:46,560 --> 00:27:48,800 EXPRESSION RESISTANT TO IT. 783 00:27:48,800 --> 00:27:50,040 THEREFORE, IN ORDER TO FURTHER 784 00:27:50,040 --> 00:27:51,600 EXPLORE THESE DIFFERENCES WE 785 00:27:51,600 --> 00:27:54,400 USED DNA BINDING EXPERIMENTS TO 786 00:27:54,400 --> 00:27:56,560 EXAMINE BOTH VASOOCCUPANCY AND 787 00:27:56,560 --> 00:27:59,880 TNF INDUCED RECRUITMENT OF RNA 788 00:27:59,880 --> 00:28:01,480 POLYMERASE 2 AND XPV. 789 00:28:01,480 --> 00:28:04,160 ON THE LEFT TOTAL RNA POL 2 790 00:28:04,160 --> 00:28:06,040 BINDING IS SHOWN RELATIVE TO 791 00:28:06,040 --> 00:28:07,440 UNSTIMULATED CONTROL CELLS, 792 00:28:07,440 --> 00:28:09,200 BINDING TO THE IL 8 PROMOTER 793 00:28:09,200 --> 00:28:10,680 SHOWN IN WHITE AND BINDING TO 794 00:28:10,680 --> 00:28:12,920 THE I KAPPA B ALPHA PROMOTER IN 795 00:28:12,920 --> 00:28:13,120 BLACK. 796 00:28:13,120 --> 00:28:14,960 YOU CAN SEE TNF STIMULATION HAS 797 00:28:14,960 --> 00:28:18,400 A MUCH LARGER EFFECT ON RNA POL 798 00:28:18,400 --> 00:28:20,640 2 RECRUITMENT TO IL 8 COMPARED 799 00:28:20,640 --> 00:28:23,600 TO THE I KAPPA BE ALPHA 800 00:28:23,600 --> 00:28:24,440 PROMOTER. 801 00:28:24,440 --> 00:28:27,360 CONSISTENT WITH IL 8 802 00:28:27,360 --> 00:28:29,920 TRANSCRIPTION IT INDUCES POLL 2 803 00:28:29,920 --> 00:28:31,480 RECRUITMENT BY MORE THAN 50%. 804 00:28:31,480 --> 00:28:34,280 THE EFFECT AT THE I KAPPA B 805 00:28:34,280 --> 00:28:36,320 ALPHA PROMOTER WAS MORE MODEST. 806 00:28:36,320 --> 00:28:38,400 NOTABLY RNA POL 2 APPEARS 807 00:28:38,400 --> 00:28:41,000 PRE-POSITIONED AT THE iKAPPA B 808 00:28:41,000 --> 00:28:43,200 ALPHA PROTOMOTORRER HAVE 809 00:28:43,200 --> 00:28:45,000 VASOSOCK PANSY IS HIGHER THAN 810 00:28:45,000 --> 00:28:46,520 THE IL 8 PROET NOER. 811 00:28:46,520 --> 00:28:48,000 THESE RESULTS WERE SIMILAR WHEN 812 00:28:48,000 --> 00:28:50,200 PROBING ACTIVATED RNA POLYMERASE 813 00:28:50,200 --> 00:28:52,400 2 DNA BINDING. 814 00:28:52,400 --> 00:28:53,120 CONSISTENT WITH DIFFERENCES 815 00:28:53,120 --> 00:28:54,720 BETWEEN THE PROMOTER STRUCTURE 816 00:28:54,720 --> 00:28:57,280 OF THESE TWO GENES, TNF ALPHA 817 00:28:57,280 --> 00:28:59,040 STIMULATION LED TO SIGNIFICANTLY 818 00:28:59,040 --> 00:29:01,360 MORE XPB RECRUITMENT TO THE IL 8 819 00:29:01,360 --> 00:29:03,360 COMPARED TO I KAPPA B ALPHA 820 00:29:03,360 --> 00:29:04,280 PROMOTER. 821 00:29:04,280 --> 00:29:06,680 SIMILAR TO RNA POL 2 IT 822 00:29:06,680 --> 00:29:08,920 SUPPRESSED TNF INDUCE 823 00:29:08,920 --> 00:29:10,920 RECRUITMENT OF XPB TO IL 8 824 00:29:10,920 --> 00:29:13,800 PROMOTE EVERY BY MORE THAN 50 -- 825 00:29:13,800 --> 00:29:16,040 PROMOTER BY 50% EFFECTS 826 00:29:16,040 --> 00:29:19,080 RECRUITMENT OF I KAPPA BE ALPHA 827 00:29:19,080 --> 00:29:19,560 WAS MUCH LESS. 828 00:29:19,560 --> 00:29:22,960 IN CONTRAST, VASOXPB OCCUPANCY 829 00:29:22,960 --> 00:29:24,600 WAS NOT SIGNIFICANTLY DIFFERENT 830 00:29:24,600 --> 00:29:26,320 BETWEEN TWO PROMOTER REGIONS. 831 00:29:26,320 --> 00:29:29,920 IN SUMMARY, S PURNO LACTONE 832 00:29:29,920 --> 00:29:33,760 BLOCKS RECRUITMENT OF ACTIVATED 833 00:29:33,760 --> 00:29:38,800 RNA POL 2 BUT NOT THE -- 834 00:29:38,800 --> 00:29:40,880 VASOOCCUPANCY OF TOTAL ACTIVATED 835 00:29:40,880 --> 00:29:43,440 PLO 2 WAS SIGNIFICANTLY HIGHER 836 00:29:43,440 --> 00:29:46,240 AT THE IL 8 PROMOTER REGION. 837 00:29:46,240 --> 00:29:49,200 THEREFORE DIFFERENCES IN VASORNA 838 00:29:49,200 --> 00:29:50,880 POL 2 OCCUPANCY APPEAR TO 839 00:29:50,880 --> 00:29:52,880 DISTINGUISH BETWEEN GENES MORE 840 00:29:52,880 --> 00:29:55,440 OR LESS PRONE TO SPURNO LACTONE 841 00:29:55,440 --> 00:29:57,040 MEDIATED DEGRADATION. 842 00:29:57,040 --> 00:30:01,680 THUS IMPLYING IT DOES NOT 843 00:30:01,680 --> 00:30:02,880 INDISCRIMINATELY IMPAIR GLOBAL 844 00:30:02,880 --> 00:30:03,720 GENE TRANSCRIPTION. 845 00:30:03,720 --> 00:30:07,840 BELOW IS A SIMPLIFIED SCHEMATIC 846 00:30:07,840 --> 00:30:09,800 OF EVENTS AT PROMOTER REGION OF 847 00:30:09,800 --> 00:30:10,840 TARGET GENE, FOLLOWING 848 00:30:10,840 --> 00:30:13,040 ACTIVATION OF CELL THE 849 00:30:13,040 --> 00:30:14,920 INFLAMMATORY TRANSCRIPTION 850 00:30:14,920 --> 00:30:16,080 FACTORS ARE RECRUITED TO THE 851 00:30:16,080 --> 00:30:16,720 RESPECTSTIVE BINDING SITES. 852 00:30:16,720 --> 00:30:19,120 THIS IS ACCOMPANIED BY 853 00:30:19,120 --> 00:30:22,360 RECRUITMENT OF THE TRANSCRIPTION 854 00:30:22,360 --> 00:30:23,160 GNASHIATION COMPLEX AND 855 00:30:23,160 --> 00:30:23,960 ACTIVATION OF GENE 856 00:30:23,960 --> 00:30:24,560 TRANSCRIPTION. 857 00:30:24,560 --> 00:30:26,680 HOWEVER, IN THE PRESENCE OF 858 00:30:26,680 --> 00:30:29,440 SPURNO LACTONE XPB AN ENTRYCAL 859 00:30:29,440 --> 00:30:31,680 SUB UNIT OF TRANSCRIPTION FACTOR 860 00:30:31,680 --> 00:30:34,400 TF 2H IS DEGRADED BY THE 861 00:30:34,400 --> 00:30:34,760 PROTEOSOME. 862 00:30:34,760 --> 00:30:36,440 THIS DOESN'T ALTER TRANSCRIPTION 863 00:30:36,440 --> 00:30:38,240 FACTOR BINDING, IT DOES RESULT 864 00:30:38,240 --> 00:30:41,520 IN DECREASE RECRUITMENT OF RNA 865 00:30:41,520 --> 00:30:44,640 POL 2 AND XPB TO THE PROMOTER 866 00:30:44,640 --> 00:30:46,280 REGION LEADING TO SUPPRESSION OF 867 00:30:46,280 --> 00:30:47,400 INFLAMMATORY GENE TRANSCRIPTION. 868 00:30:47,400 --> 00:30:49,200 NOTABLY THIS IS THE FIRST 869 00:30:49,200 --> 00:30:54,280 DETAILED CHARACTERIZATION OF MR 870 00:30:54,280 --> 00:30:57,160 INDEPENDENT MECHANISM FOR SPURNO 871 00:30:57,160 --> 00:30:57,960 LACTONE. 872 00:30:57,960 --> 00:31:01,200 IT INDUCE XPV DEGRADATION, 873 00:31:01,200 --> 00:31:02,240 EXPRESSES GENE TRANSCRIPTION AND 874 00:31:02,240 --> 00:31:04,160 IT IS A APPROPRIATE TO IT FOR 875 00:31:04,160 --> 00:31:06,280 NEW CLASS OF DRUGS THAT DEGRADES 876 00:31:06,280 --> 00:31:07,320 SPECIFIC PROTEIN TARGETS WHICH 877 00:31:07,320 --> 00:31:08,440 MAYBE USEFUL STRATEGY FOR 878 00:31:08,440 --> 00:31:10,480 MODULATING INFLAMMATION. 879 00:31:10,480 --> 00:31:12,600 WHETHER CLINICALLY RELEVANT 880 00:31:12,600 --> 00:31:15,480 CONCENTRATION OF IT LEAD TO XPV 881 00:31:15,480 --> 00:31:16,600 DEGRADATION AND CONTRIBUTE TO 882 00:31:16,600 --> 00:31:18,720 CLINICAL BENEFITS REMAINS 883 00:31:18,720 --> 00:31:19,120 UNKNOWN. 884 00:31:19,120 --> 00:31:21,520 NEXT I WILL DISCUSS HOW OUR LAB 885 00:31:21,520 --> 00:31:22,840 IS LEVERAGING CELLULAR MODELS OF 886 00:31:22,840 --> 00:31:25,600 PAH AS A TOOL FOR DISSECTING THE 887 00:31:25,600 --> 00:31:26,800 PATHOBIOLOGY UNDERLYING LUNG 888 00:31:26,800 --> 00:31:30,360 VESSEL INFLAMMATION. 889 00:31:30,360 --> 00:31:31,440 INVESTIGATING MECHANISMS THAT 890 00:31:31,440 --> 00:31:33,800 CONTRIBUTE TO VASCULAR MODEL 891 00:31:33,800 --> 00:31:35,320 ROCK HAMPERD BY INABILITY TO 892 00:31:35,320 --> 00:31:36,960 SAFELY OBTAIN LUNG TISSUE DURING 893 00:31:36,960 --> 00:31:39,480 COURSE OF PATIENT DISEASE. 894 00:31:39,480 --> 00:31:41,480 FOR INSTANCE THE HISTOPATHOLOGIC 895 00:31:41,480 --> 00:31:43,960 SAMPLE HERE CAN BE OBTAINED FROM 896 00:31:43,960 --> 00:31:45,120 EITHER EXPLANTED LUNGS AT TIME 897 00:31:45,120 --> 00:31:48,320 OF LUNG TRANSPLANT OR AUTOPSY. 898 00:31:48,320 --> 00:31:49,880 NOTABLY THE PREDICTED 899 00:31:49,880 --> 00:31:50,880 CONSEQUENCEs OF ALL KNOWN 900 00:31:50,880 --> 00:31:53,360 MUTATIONS IN PAH ASSOCIATED 901 00:31:53,360 --> 00:31:55,120 GENES IS LOSS OF FUNCTION. 902 00:31:55,120 --> 00:31:57,080 THEREFORE WE HAVE TAKEN A 903 00:31:57,080 --> 00:31:59,280 MOLECULAR APPROACH UTILIZING 904 00:31:59,280 --> 00:32:00,840 SMALL INTERFERING RNA 905 00:32:00,840 --> 00:32:03,200 TRANSFECTION TO MIMIC PAH 906 00:32:03,200 --> 00:32:04,920 ASSOCIATED MOLECULAR DEFECTS. 907 00:32:04,920 --> 00:32:06,440 CURRENTLY WE ARE INVESTIGATING A 908 00:32:06,440 --> 00:32:07,480 NUMBER OF DIFFERENT CELL MODELS 909 00:32:07,480 --> 00:32:09,800 IN OUR LAB, AND TODAY I WILL 910 00:32:09,800 --> 00:32:12,000 HIGHLIGHT ONE MODEL IN 911 00:32:12,000 --> 00:32:16,000 PARTICULAR BASED ON SILENCING 912 00:32:16,000 --> 00:32:19,240 CAV 1 IN PRIMARY HUMAN ARTERY 913 00:32:19,240 --> 00:32:19,720 ENDOTHELIAL CELLS. 914 00:32:19,720 --> 00:32:21,880 LOSS OF FUNCTION MUTATIONS ARE A 915 00:32:21,880 --> 00:32:25,040 RARE HEREDITARY CAUSE OF PAH. 916 00:32:25,040 --> 00:32:26,480 HOWEVER, SUGGESTING AN EVEN 917 00:32:26,480 --> 00:32:29,360 BROADER ROLE IN DISEASE 918 00:32:29,360 --> 00:32:31,240 PATHOGENESIS, WILD TYPE CAV 1 919 00:32:31,240 --> 00:32:33,560 PROTEIN EXPRESSION IS REDUCED IN 920 00:32:33,560 --> 00:32:34,840 LUNG VASCULAR LESION OF PATIENTS 921 00:32:34,840 --> 00:32:36,600 WITH IDIOPATHIC AND DISEASE 922 00:32:36,600 --> 00:32:39,040 ASSOCIATED PAH. 923 00:32:39,040 --> 00:32:42,800 AS YOU CAN SEE GENE SILENCING 924 00:32:42,800 --> 00:32:45,000 SIGNIFICANTLY REDUCED CAV 1 925 00:32:45,000 --> 00:32:46,560 EXPRESSION IN HUMAN PULMONARY 926 00:32:46,560 --> 00:32:48,080 ARTERY ENDOTHELIAL CELLS. 927 00:32:48,080 --> 00:32:49,760 RELATIVE mRNA SHOWN ON THE 928 00:32:49,760 --> 00:32:51,520 LEFT AND PROTEIN EXPRESSION IS 929 00:32:51,520 --> 00:32:53,800 SHOWN ON THE RIGHT. 930 00:32:53,800 --> 00:32:56,240 IN OUR MODEL CAV 1 DEFICIENCY 931 00:32:56,240 --> 00:32:58,440 PRODUCED A DYSFUNCTIONAL PAH 932 00:32:58,440 --> 00:33:00,320 LIKE ENDSTHELIAL PHENOTYPE, 933 00:33:00,320 --> 00:33:01,440 SHOWN HERE ON THE LEFT YOU CAN 934 00:33:01,440 --> 00:33:03,880 SEE THAT COMPARED TO CONTROL S 935 00:33:03,880 --> 00:33:06,880 RNA TRANSFECTED CELLS IN RED CAV 936 00:33:06,880 --> 00:33:08,680 1 SILENCE CELLS WERE HIGHLY 937 00:33:08,680 --> 00:33:09,280 PROLIFERATIVE. 938 00:33:09,280 --> 00:33:11,280 LIKEWISE WHEN COMPARED TO NORMAL 939 00:33:11,280 --> 00:33:13,640 CELL MIGRATION OVER 72 HOURS IN 940 00:33:13,640 --> 00:33:17,080 CULTURE, CAV 1 LOSS PRODUCED 941 00:33:17,080 --> 00:33:19,160 HYPERMIGRATORY PHENOTYPE. 942 00:33:19,160 --> 00:33:20,520 NEXT GENOME WIDE EXPRESSION 943 00:33:20,520 --> 00:33:22,840 PROFILING WAS PERFORMED TO 944 00:33:22,840 --> 00:33:25,880 UNBIASLY ASSESS IMPACT OF CAV 1 945 00:33:25,880 --> 00:33:27,480 DEFICIENCY ON GENE TRANSCRIPTION 946 00:33:27,480 --> 00:33:29,320 IN PULMONARY ARTERY ENDOTHELIAL 947 00:33:29,320 --> 00:33:30,520 CELLS. 948 00:33:30,520 --> 00:33:31,960 CAV 1 SILENCE CELLS SHOWED 949 00:33:31,960 --> 00:33:33,480 STRONG ENRICHMENT FOR THE 950 00:33:33,480 --> 00:33:35,200 ANTIVIRAL RESPONSE AND SYSTEMIC 951 00:33:35,200 --> 00:33:35,720 AUTO-IMMUNITY. 952 00:33:35,720 --> 00:33:38,040 IN ADDITION, INTERFERON 953 00:33:38,040 --> 00:33:39,280 SIGNALING PREDICTED TO BE 954 00:33:39,280 --> 00:33:41,520 ACTIVATING CAV 1 SILENCE CELLS 955 00:33:41,520 --> 00:33:43,360 WAS IDENTIFIED AS THE TOP 956 00:33:43,360 --> 00:33:46,400 CANONICAL PATHWAY. 957 00:33:46,400 --> 00:33:47,840 EXPRESSION OF A STRONG 958 00:33:47,840 --> 00:33:50,200 INTERFERON GENE SIGNATURE IN OUR 959 00:33:50,200 --> 00:33:51,840 TRANSCRIPTOMIC ANALYSIS 960 00:33:51,840 --> 00:33:52,880 INDICATED ENDOTHELIAL 961 00:33:52,880 --> 00:33:54,760 DYSFUNCTION FROM CAV 1 LOSS 962 00:33:54,760 --> 00:33:57,120 MIGHT BE A DIRECT CONSEQUENCE OF 963 00:33:57,120 --> 00:33:59,600 CONSTITUTIVE INTERFERON 964 00:33:59,600 --> 00:34:00,520 INFLAMMATORY RESPONSE. 965 00:34:00,520 --> 00:34:02,000 CONSISTENT WITH THIS HYPOTHESIS 966 00:34:02,000 --> 00:34:04,320 YOU SEE THAT TOTAL AND 967 00:34:04,320 --> 00:34:05,680 PHOSPHORYLATED STAT 1 LEVELS ARE 968 00:34:05,680 --> 00:34:08,120 INCREASED IN CAV 1 CELLS, 969 00:34:08,120 --> 00:34:10,120 THERE'S INCREASE DNA BINDING OF 970 00:34:10,120 --> 00:34:11,840 BOTH INTERFERON REGULATORY 971 00:34:11,840 --> 00:34:14,160 FACTORS 3 AND 7, AS WELL AS 972 00:34:14,160 --> 00:34:16,040 INDUCTION OF CANONICAL 973 00:34:16,040 --> 00:34:17,280 INTERFERON TARGET GENES 974 00:34:17,280 --> 00:34:19,200 INCLUDING CXCL 10. 975 00:34:19,200 --> 00:34:20,240 COLLECTIVELY THESE RESULTS 976 00:34:20,240 --> 00:34:22,720 DEMONSTRATE CAV 1 LOSS 977 00:34:22,720 --> 00:34:24,400 CONSTITUTIVELY ACTIVATES 978 00:34:24,400 --> 00:34:25,800 ENDOGENOUS JAK STAT INTERFERON 979 00:34:25,800 --> 00:34:26,840 SIGNALING. 980 00:34:26,840 --> 00:34:28,960 IMPORTANTLY, IN PAH PATIENTS 981 00:34:28,960 --> 00:34:32,600 WITH PATHOGENIC CAV 1 MUTATIONS, 982 00:34:32,600 --> 00:34:35,040 ELEVATED CXCL 10 LEVELS AND 983 00:34:35,040 --> 00:34:39,200 THEIR CELLS MIRRORED THE 984 00:34:39,200 --> 00:34:40,960 PHENOTYPIC MOLECULAR FEATURES 985 00:34:40,960 --> 00:34:41,720 AND SILENCE CELLS FROM OUR 986 00:34:41,720 --> 00:34:42,120 MODEL. 987 00:34:42,120 --> 00:34:44,680 IN SUPPORT OF A BROADER ROLE FOR 988 00:34:44,680 --> 00:34:47,200 CAV 1 DEFICIENCY, ASSOCIATED 989 00:34:47,200 --> 00:34:48,560 INTERFERON ACTIVATION, WE 990 00:34:48,560 --> 00:34:50,480 PERFORMED IMMUNOFLUORESCENCE 991 00:34:50,480 --> 00:34:51,800 STAINING OF EXPLANTED LUNG 992 00:34:51,800 --> 00:34:53,360 TISSUE FROM PATIENTS WITH 993 00:34:53,360 --> 00:34:55,000 IDIOPATHIC PAH. 994 00:34:55,000 --> 00:34:57,080 ON THE TOP THE ARROWS INDICATE 995 00:34:57,080 --> 00:34:59,040 GREEN STAINING FOR CAV 1 996 00:34:59,040 --> 00:35:00,520 EXPRESSION IN ENDSTHELIAL LINING 997 00:35:00,520 --> 00:35:02,800 OF A NORMAL VESSEL. 998 00:35:02,800 --> 00:35:06,320 BELOW IN IPAH TISSUE I'M 999 00:35:06,320 --> 00:35:08,120 OUTLINING THE ENDOTHELIAL LINING 1000 00:35:08,120 --> 00:35:11,200 OF ABNORMAL VESSEL IN YELLOW AND 1001 00:35:11,200 --> 00:35:12,760 IN CONTRAST TO HEALTHY YOU SEE 1002 00:35:12,760 --> 00:35:14,920 SIGNIFICANT REDUCTION OF CAV 1 1003 00:35:14,920 --> 00:35:16,880 STAINENING THE LINING OF THIS 1004 00:35:16,880 --> 00:35:18,520 IPAH VESSEL. 1005 00:35:18,520 --> 00:35:19,880 LIKEWISE, THERE IS ABSENCE OF 1006 00:35:19,880 --> 00:35:22,160 RED STAINING FOR PHOSPHORYLATED 1007 00:35:22,160 --> 00:35:25,160 STAT 1 IN HEALTHY ENDOTHELIUM, 1008 00:35:25,160 --> 00:35:26,360 STAT 1 PHOSPHORYLATION IS 1009 00:35:26,360 --> 00:35:29,000 SIGNIFICANTLY HIGHER IN IPH 1010 00:35:29,000 --> 00:35:30,120 ENDOTHELIAL CELLS AS INDICATED 1011 00:35:30,120 --> 00:35:33,440 BY THE RED STAINING BELOW. 1012 00:35:33,440 --> 00:35:36,080 NEXT THE JAK STAT INTERFERON 1013 00:35:36,080 --> 00:35:37,800 PATHWAY EXPLORED POTENTIAL 1014 00:35:37,800 --> 00:35:40,120 THERAPEUTIC TARGET IN PAH 1015 00:35:40,120 --> 00:35:42,760 ASSOCIATED WITH CAV 1 LOSS. 1016 00:35:42,760 --> 00:35:44,680 USING CLINICALLY ACHIEVABLE 1017 00:35:44,680 --> 00:35:46,000 CONCENTRATIONS THREE CURRENTLY 1018 00:35:46,000 --> 00:35:47,480 AVAILABLE JACK INHIBITORS NOT 1019 00:35:47,480 --> 00:35:50,760 ONLY BLOCK STAT 1 ACTIVATION AND 1020 00:35:50,760 --> 00:35:52,200 CXCL 10 EXPRESSION BUT ALSO 1021 00:35:52,200 --> 00:35:54,760 BLOCK CELL PROLIFERATION AND 1022 00:35:54,760 --> 00:35:57,400 MIGRATION IN CAV 1 SILENCE 1023 00:35:57,400 --> 00:35:57,760 CELLS. 1024 00:35:57,760 --> 00:36:01,320 SO TO SUMMARIZE, CAV 1 1025 00:36:01,320 --> 00:36:03,400 SUFFICIENCY ELICITS INFLAMMATORY 1026 00:36:03,400 --> 00:36:05,200 RESPONSE THAT RESULTS IN A PAH 1027 00:36:05,200 --> 00:36:07,840 LIKE CELLULAR PHENOTYPE. 1028 00:36:07,840 --> 00:36:10,000 INHIBITING JAK STAT REVERSED 1029 00:36:10,000 --> 00:36:11,240 DYSFUNCTIONAL ENDSTHELIAL 1030 00:36:11,240 --> 00:36:14,200 PHENOTYPE ASSOCIATED WITH CAV 1 1031 00:36:14,200 --> 00:36:14,880 LOSS IN VITRO. 1032 00:36:14,880 --> 00:36:15,880 IN EXPERIMENTS I DIDN'T HAVE 1033 00:36:15,880 --> 00:36:17,840 TIME TO SHARE, INTERESTINGLY 1034 00:36:17,840 --> 00:36:19,600 EXOGENOUS INTERFERON STIMULATION 1035 00:36:19,600 --> 00:36:22,040 ALONE REDUCES CAV 1 PROTEIN 1036 00:36:22,040 --> 00:36:23,360 LEVELS IN OTHERWISE NORMAL 1037 00:36:23,360 --> 00:36:24,720 PULMONARY ARTERY ENDOTHELIAL 1038 00:36:24,720 --> 00:36:26,000 CELLS. 1039 00:36:26,000 --> 00:36:28,440 SUGGESTING THAT CAV 1 1040 00:36:28,440 --> 00:36:29,640 INSUFFICIENCY MAY CONTRIBUTE TO 1041 00:36:29,640 --> 00:36:32,800 DEVELOPMENT OF AUTOIMMUNE AND 1042 00:36:32,800 --> 00:36:34,440 AUTOINFLAMMATORY DISEASE 1043 00:36:34,440 --> 00:36:34,880 ASSOCIATE CANNED PH. 1044 00:36:34,880 --> 00:36:36,760 THEREFORE CELLULAR MODELS OF PAH 1045 00:36:36,760 --> 00:36:38,520 ASSOCIATED MOLECULAR DEFECTS CAN 1046 00:36:38,520 --> 00:36:40,200 BE LEVERAGED TO FURTHER OUR 1047 00:36:40,200 --> 00:36:42,080 UNDERSTANDING OF DISEASE 1048 00:36:42,080 --> 00:36:44,520 PATHOBIOLOGY, AS WELL AS A TOOL 1049 00:36:44,520 --> 00:36:47,520 TO IDENTIFY THERAPEUTIC TARGETS. 1050 00:36:47,520 --> 00:36:49,840 I WILL NOW PASS THE BATON BACK 1051 00:36:49,840 --> 00:36:51,640 TO YOU MIKE SO YOU CAN EXPLAIN 1052 00:36:51,640 --> 00:36:54,040 HOW WE TRANSLATED OUR INTEREST 1053 00:36:54,040 --> 00:36:55,600 AND INFLAMMATION INTO OUR 1054 00:36:55,600 --> 00:36:57,000 CLINICAL STUDIES IN IN PAH 1055 00:36:57,000 --> 00:37:00,720 PATIENTS. 1056 00:37:00,720 --> 00:37:01,600 >> THANK YOU. 1057 00:37:01,600 --> 00:37:03,480 THE TRANSLATIONAL ARM OF THE NIH 1058 00:37:03,480 --> 00:37:05,960 CLINICAL CENTER PAH SECTION HAS 1059 00:37:05,960 --> 00:37:07,360 SYNERGIZED WITH AND BUILT UPON 1060 00:37:07,360 --> 00:37:09,680 DR. ELINOFF'S BENCH 1061 00:37:09,680 --> 00:37:11,120 INVESTIGATIONS TO FURTHER 1062 00:37:11,120 --> 00:37:12,800 ELUCIDATE INFLAMMATORY PROCESSES 1063 00:37:12,800 --> 00:37:15,800 IN PAH WITH THE ULTIMATE GOAL OF 1064 00:37:15,800 --> 00:37:17,240 INTERVENING THERAPEUTICALLY. 1065 00:37:17,240 --> 00:37:19,600 IN COLLABORATION WITH THE NIH 1066 00:37:19,600 --> 00:37:21,600 CENTER FOR HUMAN IMMUNOLOGY, WE 1067 00:37:21,600 --> 00:37:24,480 ALSO LOOKED FOR EVIDENCE OF 1068 00:37:24,480 --> 00:37:25,840 ELEVATED SITE -- INFLAMMATORY 1069 00:37:25,840 --> 00:37:27,720 CYTOKINES IN OUR PAH POPULATION. 1070 00:37:27,720 --> 00:37:30,520 SHOWN ON THIS SLIDE ARE SERUM 1071 00:37:30,520 --> 00:37:34,040 CONCENTRATIONS, OF IL 6, IL 8, 1072 00:37:34,040 --> 00:37:36,680 TNF ALPHA AND INTERFERON GAMMA. 1073 00:37:36,680 --> 00:37:38,280 HEALTHY CONTROLS ARE SHOWN IN 1074 00:37:38,280 --> 00:37:39,840 BLACK AND PH PATIENTS IN RED. 1075 00:37:39,840 --> 00:37:41,840 AS YOU CAN SEE THERE IS 1076 00:37:41,840 --> 00:37:43,600 SIGNIFICANT ELEVATION OF ALL 1077 00:37:43,600 --> 00:37:45,800 SERUM CYTOKINE LEVELS IN PH 1078 00:37:45,800 --> 00:37:47,280 PATIENTS COMPARED TO HEALTHY 1079 00:37:47,280 --> 00:37:48,640 CONTROLS PROVIDING EVIDENCE OF 1080 00:37:48,640 --> 00:37:50,240 SYSTEMIC INFLAMMATION IN IF 1081 00:37:50,240 --> 00:37:52,400 PULMONARY ARTERIAL HYPERTENSION. 1082 00:37:52,400 --> 00:37:54,520 NEXT IN COLLABORATION WITH THE 1083 00:37:54,520 --> 00:37:56,280 BIOMEDICAL AND METABOLIC IMAGING 1084 00:37:56,280 --> 00:37:58,160 BRANCH, WE ASSESSED CORONARY 1085 00:37:58,160 --> 00:37:59,880 PLAQUE BURDEN AS ANOTHER MARKER 1086 00:37:59,880 --> 00:38:01,840 OF SYSTEMIC INFLAMMATION. 1087 00:38:01,840 --> 00:38:04,360 THIS CARDIAC CT IMAGE IS FROM A 1088 00:38:04,360 --> 00:38:06,280 3-D GLOBAL RECONSTRUCTED 1089 00:38:06,280 --> 00:38:07,320 PROJECTION OF THE HEART SHOWN IN 1090 00:38:07,320 --> 00:38:10,320 THE CENTER IS AORTIC VALVE WITH 1091 00:38:10,320 --> 00:38:11,960 PROJECTING CORONARY ARTERIES 1092 00:38:11,960 --> 00:38:12,920 COLOR CODED. 1093 00:38:12,920 --> 00:38:15,000 TO ASSESS CORONARY PLAQUE 1094 00:38:15,000 --> 00:38:16,800 BURDEN, WE DIVIDE THE CORONARY 1095 00:38:16,800 --> 00:38:19,560 SYSTEM INTO 16 SEGMENTS AND 1096 00:38:19,560 --> 00:38:22,320 DETERMINE THE PERCENT OF THE 16 1097 00:38:22,320 --> 00:38:25,600 SEGMENTS WITH EVIDENCE OF 1098 00:38:25,600 --> 00:38:26,240 ATHEROSCLEROSIS. 1099 00:38:26,240 --> 00:38:27,320 PH PATIENTS ARE MATCHED TO 1100 00:38:27,320 --> 00:38:29,200 CONTROLS BY RISK FOR CORONARY 1101 00:38:29,200 --> 00:38:31,640 ARTERY DISEASE USING AGE, 1102 00:38:31,640 --> 00:38:35,400 GENDER, AND FRAMINGHAM SCORE. 1103 00:38:35,400 --> 00:38:37,600 THE BOX AND WHISKER PLOTS SHOW 1104 00:38:37,600 --> 00:38:39,320 PH PATIENTS IN RED BOX HAVE 1105 00:38:39,320 --> 00:38:41,480 SIGNIFICANTLY GREATER BURDEN OF 1106 00:38:41,480 --> 00:38:42,520 CORONARY PLAQUE COMPARED TO 1107 00:38:42,520 --> 00:38:44,080 HEALTHY INDIVIDUALS SHOWN IN THE 1108 00:38:44,080 --> 00:38:45,000 BLUE BOX. 1109 00:38:45,000 --> 00:38:46,360 AGAIN, EVIDENCE OF SYSTEMIC 1110 00:38:46,360 --> 00:38:48,280 INFLAMMATION. 1111 00:38:48,280 --> 00:38:51,040 IN OUR COHORT WE ALSO COMPARED 1112 00:38:51,040 --> 00:38:54,160 AND LOOK AT BLOOD MONONUCLEAR 1113 00:38:54,160 --> 00:38:55,400 CELL GENE EXPRESSION PROFILING 1114 00:38:55,400 --> 00:38:57,640 AMONG HEALTHY AND PULMONARY 1115 00:38:57,640 --> 00:38:59,280 ARTERIAL HYPERTENSION PATIENTS. 1116 00:38:59,280 --> 00:39:01,800 THIS HEAT MAP DEPICTS A 1117 00:39:01,800 --> 00:39:03,160 SUBSTANCETIVE GENES FROM PH LIST 1118 00:39:03,160 --> 00:39:06,280 THAT ANNOTATED TO INFLAMMATORY 1119 00:39:06,280 --> 00:39:06,760 RESPONSE. 1120 00:39:06,760 --> 00:39:08,720 EACH COLUMN REPRESENTS A 1121 00:39:08,720 --> 00:39:09,520 SUBJECT. 1122 00:39:09,520 --> 00:39:11,720 TEN HEALTHY SUBJECTS, AND TEN 1123 00:39:11,720 --> 00:39:13,360 PAH PATIENTS. 1124 00:39:13,360 --> 00:39:15,720 EACH ROW REPRESENTS AN 1125 00:39:15,720 --> 00:39:17,240 INFLAMMATORY RESPONSE GENE. 1126 00:39:17,240 --> 00:39:20,000 RED INDICATES OVEREXPRESSION AND 1127 00:39:20,000 --> 00:39:21,600 GREEN INDICATES UNDEREXPRESSION 1128 00:39:21,600 --> 00:39:23,000 OF THE TRANSCRIPT. 1129 00:39:23,000 --> 00:39:25,400 LISTED ALONG THE Y AXIS ARE 1130 00:39:25,400 --> 00:39:27,000 EXAMPLES OF INFLAMMATORY 1131 00:39:27,000 --> 00:39:29,200 RESPONSE GENES FROM OUR DATA 1132 00:39:29,200 --> 00:39:31,080 SET, SUCH AS INTEGRINS, 1133 00:39:31,080 --> 00:39:33,440 INTERLEUKINS AND P SELECTED. 1134 00:39:33,440 --> 00:39:36,080 THE PH PATIENTS COMPARED TO THE 1135 00:39:36,080 --> 00:39:38,120 HEALTHY CONTROLS SHOW CLEAR 1136 00:39:38,120 --> 00:39:39,800 OVEREXPRESSION OF INFLAMMATORY 1137 00:39:39,800 --> 00:39:42,760 RESPONSE GENES AS INDICATED BY 1138 00:39:42,760 --> 00:39:46,320 THE CONSIDERABLE BRIGHT RED 1139 00:39:46,320 --> 00:39:47,560 COLOR OF THEIR PANEL. 1140 00:39:47,560 --> 00:39:50,360 THERE EXISTS ABNORMAL 1141 00:39:50,360 --> 00:39:52,360 INFLAMMATORY RESPONSE IN BLOOD 1142 00:39:52,360 --> 00:39:55,400 MONONUCLEAR CELLS OF PAH 1143 00:39:55,400 --> 00:39:55,840 PATIENTS. 1144 00:39:55,840 --> 00:39:58,400 SUBSEQUENTLY WE COMPARED BLOOD 1145 00:39:58,400 --> 00:39:59,520 MONONUCLEAR CELL GENE EXPRESSION 1146 00:39:59,520 --> 00:40:00,680 PROFILING AMONG THE DIFFERENT 1147 00:40:00,680 --> 00:40:02,920 GROUPS OF PAH PATIENTS AND 1148 00:40:02,920 --> 00:40:03,840 HEALTHY CONTROLS. 1149 00:40:03,840 --> 00:40:07,480 SHOWN HERE ARE FOUR GROUPS, 1150 00:40:07,480 --> 00:40:08,200 IDIOPATHIC PULMONARY 1151 00:40:08,200 --> 00:40:09,720 HYPERTENSION, CONNECTIVE TISSUE 1152 00:40:09,720 --> 00:40:10,800 DISEASE ASSOCIATED PULMONARY 1153 00:40:10,800 --> 00:40:13,400 ARTERIAL HYPERTENSION, AND OTHER 1154 00:40:13,400 --> 00:40:14,800 ETIOLOGIES OF PULMONARY ARTERIAL 1155 00:40:14,800 --> 00:40:16,560 HYPERTENSION AND HEALTHY 1156 00:40:16,560 --> 00:40:17,120 VOLUNTEER GROUP. 1157 00:40:17,120 --> 00:40:19,320 ONCE AGAIN IN THE HEAT MAP, RED 1158 00:40:19,320 --> 00:40:21,240 INDICATES INCREASE EXPRESSION 1159 00:40:21,240 --> 00:40:22,960 AND THIS TIME BLUE INDICATES 1160 00:40:22,960 --> 00:40:25,200 DECREASED EXPRESSION. 1161 00:40:25,200 --> 00:40:27,560 EACH COLUMN REPRESENTS A 1162 00:40:27,560 --> 00:40:29,240 DIFFERENT PERSON AS ILLUSTRATED 1163 00:40:29,240 --> 00:40:31,600 BY THE BLACK VERTICAL BOX. 1164 00:40:31,600 --> 00:40:33,800 EACH ROW REPRESENTS A DIFFERENT 1165 00:40:33,800 --> 00:40:35,720 BLOOD INFLAMMATORY PROTEIN AS 1166 00:40:35,720 --> 00:40:37,720 SHOWN BY THE BLACK HORIZONTAL 1167 00:40:37,720 --> 00:40:38,760 BOX. 1168 00:40:38,760 --> 00:40:40,400 PLEASE APPRECIATE HOW MUCH 1169 00:40:40,400 --> 00:40:42,720 REDDER IT IS FOR THOSE WITH PAH 1170 00:40:42,720 --> 00:40:44,000 COMPARED TO ALL THE BLUE YOU SEE 1171 00:40:44,000 --> 00:40:45,840 ON THE RIGHT SIDE WHERE THE 1172 00:40:45,840 --> 00:40:48,120 HEALTHY VOLUNTEERS ARE LOCATED. 1173 00:40:48,120 --> 00:40:49,920 THUS THE STUDY DETECTED AN 1174 00:40:49,920 --> 00:40:51,720 ABNORMAL INFLAMMATORY RESPONSE 1175 00:40:51,720 --> 00:40:54,040 IN THE BLOOD OF PH PATIENTS EVEN 1176 00:40:54,040 --> 00:40:58,000 WHEN DIVIDED BY SPECIFIC 1177 00:40:58,000 --> 00:40:58,600 DIFFERING ETIOLOGIES. 1178 00:40:58,600 --> 00:41:01,480 WE ALSO PERFORM AD META ANALYSIS 1179 00:41:01,480 --> 00:41:03,040 OF BLOOD GENOME WIDE EXPRESSION 1180 00:41:03,040 --> 00:41:04,760 PROFILING STUDIES IN PULMONARY 1181 00:41:04,760 --> 00:41:06,000 ARTERIAL HYPERTENSION. 1182 00:41:06,000 --> 00:41:08,440 WE FOUND INFLAMMATORY PATHWAYS 1183 00:41:08,440 --> 00:41:11,520 TO BE OVERREPRESENTED AMONG 1184 00:41:11,520 --> 00:41:13,240 BIOFUNCTIONS, AS SHOWN ON THE 1185 00:41:13,240 --> 00:41:15,360 LEFT AND CANONICAL PATHWAYS 1186 00:41:15,360 --> 00:41:18,000 SHOWN ON THE RIGHT. 1187 00:41:18,000 --> 00:41:19,240 RECENTLY IN COLLABORATION WITH 1188 00:41:19,240 --> 00:41:20,880 THE LABORATORY OF APPLIED 1189 00:41:20,880 --> 00:41:22,720 PRECISION OMICS, WE INVESTIGATED 1190 00:41:22,720 --> 00:41:25,360 THE CONCENTRATIONS OF CELL FREE 1191 00:41:25,360 --> 00:41:27,960 DNA IN PATIENTS WITH IDIOPATHIC 1192 00:41:27,960 --> 00:41:30,920 CONNECTIVE TISSUE DISEASE AND 1193 00:41:30,920 --> 00:41:32,160 HYPERTENSION ASSOCIATED PAH. 1194 00:41:32,160 --> 00:41:34,320 COMPARED TO HEALTHY VOLUNTEER 1195 00:41:34,320 --> 00:41:35,760 PLASMA, CELL FREE DNA WAS 1196 00:41:35,760 --> 00:41:38,400 ELEVATED IN IDIOPATHIC PULMONARY 1197 00:41:38,400 --> 00:41:39,760 HYPERTENSION, CONNECTIVE TISSUE 1198 00:41:39,760 --> 00:41:41,400 DISEASE ASSOCIATED PULMONARY 1199 00:41:41,400 --> 00:41:44,840 ARTERIAL HYPERTENSION. 1200 00:41:44,840 --> 00:41:47,640 PORTAL HYPERTENSION ASSOCIATED 1201 00:41:47,640 --> 00:41:49,080 PAH AND ALL HE WILL VAGUES WERE 1202 00:41:49,080 --> 00:41:49,480 SIGNIFICANT. 1203 00:41:49,480 --> 00:41:52,960 WE USE BISULFIDE SEQUENCES AND 1204 00:41:52,960 --> 00:41:54,720 QUESTION CONVOLUTION ALGORITHM 1205 00:41:54,720 --> 00:41:56,840 TO MAP TISSUE SPECIFIC SOURCES 1206 00:41:56,840 --> 00:41:58,960 OF CELL FREE DNA. 1207 00:41:58,960 --> 00:42:00,520 THE INDIVIDUALS ARE CODE AID 1208 00:42:00,520 --> 00:42:02,720 CROSS THE DOP ROW, WITH HELL -- 1209 00:42:02,720 --> 00:42:04,520 TOP ROW WITH HEALTHY BEING 1210 00:42:04,520 --> 00:42:06,280 PURPLE COLOR AND PH REPRESENTED 1211 00:42:06,280 --> 00:42:08,200 BY GREEN YELLOW AND RED. 1212 00:42:08,200 --> 00:42:10,400 WITH RED HAVING MOST SEVERE 1213 00:42:10,400 --> 00:42:12,320 PULMONARY ARTERIAL HYPERTENSION. 1214 00:42:12,320 --> 00:42:14,560 ALONG THE RIGHT IN A VERTICAL 1215 00:42:14,560 --> 00:42:16,240 COLUMN ARE VARIOUS TISSUES THAT 1216 00:42:16,240 --> 00:42:17,800 TURN OVER AND RELEASE CELL FREE 1217 00:42:17,800 --> 00:42:19,400 DNA. 1218 00:42:19,400 --> 00:42:22,200 HERE IS THE HEAT MAP, RED 1219 00:42:22,200 --> 00:42:23,520 IMPLIES RELATIVELY INCREASED 1220 00:42:23,520 --> 00:42:25,400 CELL FREE DNA COPY, AND BLUE 1221 00:42:25,400 --> 00:42:27,640 IMPLIES RELATIVELY DECREASED 1222 00:42:27,640 --> 00:42:29,120 CELL FREE DNA COPIES. 1223 00:42:29,120 --> 00:42:32,320 ONCE AGAIN, EACH COLUMN IS AN 1224 00:42:32,320 --> 00:42:33,440 INDIVIDUAL PERSON. 1225 00:42:33,440 --> 00:42:34,560 EACH ROW REPRESENTS A TISSUE 1226 00:42:34,560 --> 00:42:36,360 TYPE. 1227 00:42:36,360 --> 00:42:39,160 INSIDE THESE HORIZONTAL BLACK 1228 00:42:39,160 --> 00:42:44,120 BOXES I HIGHLIGHTED ROWS FROM 1229 00:42:44,120 --> 00:42:47,560 CELL FREE DNA NEUTROPHILS, AND 1230 00:42:47,560 --> 00:42:48,400 NATURAL KILLER CELLS. 1231 00:42:48,400 --> 00:42:52,560 ON THE RIGHT IN THE VET PALCAL 1232 00:42:52,560 --> 00:42:54,520 BLACK BOX IS HEALTHY CONTROLS 1233 00:42:54,520 --> 00:42:55,920 NOTE PURPLE ON THE TOP. 1234 00:42:55,920 --> 00:42:57,920 ON THE LEFT INSIDE THE VERTICAL 1235 00:42:57,920 --> 00:43:00,600 BLACK BOX IS MOSTLY PATIENT, 1236 00:43:00,600 --> 00:43:01,600 NOTE RED YELLOW AND GREEN AT THE 1237 00:43:01,600 --> 00:43:01,960 TOP. 1238 00:43:01,960 --> 00:43:04,000 NOTICE THE PA PATIENTS 1239 00:43:04,000 --> 00:43:05,960 PARTICULARLY THE HIGH RISK 1240 00:43:05,960 --> 00:43:07,680 GROUP, TEND TO BE REDDER 1241 00:43:07,680 --> 00:43:09,440 INDICATING THAT PART OF THE 1242 00:43:09,440 --> 00:43:11,600 INCREASED CELL FREE DNA COPIES 1243 00:43:11,600 --> 00:43:12,960 IN PULMONARY ARTERIAL 1244 00:43:12,960 --> 00:43:14,560 HYPERTENSION PATIENTS IS COMING 1245 00:43:14,560 --> 00:43:17,840 FROM INFLAMMATORY CELLS. 1246 00:43:17,840 --> 00:43:20,000 NEXT PAIR WISE COMPARISONS WERE 1247 00:43:20,000 --> 00:43:23,240 DONE BETWEEN PH PATIENTS AND 1248 00:43:23,240 --> 00:43:24,880 HEALTHY CONTROLS WHERE CELL 1249 00:43:24,880 --> 00:43:26,920 SPECIFIC DNA CONCENTRATIONS 1250 00:43:26,920 --> 00:43:29,240 RELATED TO NEUTROPHILS, 1251 00:43:29,240 --> 00:43:31,040 MONOCYTES AND NATURAL KILLER 1252 00:43:31,040 --> 00:43:31,520 CELLS. 1253 00:43:31,520 --> 00:43:33,480 DATA ARE DISPLAYED AS A MEAN 1254 00:43:33,480 --> 00:43:35,760 PLUS OR MINUS STANDARD DEVIATION 1255 00:43:35,760 --> 00:43:38,160 OF THE LOG 10 TRANSFORM VALUES. 1256 00:43:38,160 --> 00:43:40,160 PATIENTS ARE DIVIDED TO THREE 1257 00:43:40,160 --> 00:43:42,960 RISK GROUPS, BASED ON REVEAL 2.0 1258 00:43:42,960 --> 00:43:45,760 SCORE, LOW, MEDIUM AND HIGH. 1259 00:43:45,760 --> 00:43:48,600 PAIR WISE COMPARISONS BETWEEN 1260 00:43:48,600 --> 00:43:51,600 LOW RISK PH PATIENTS AND HEALTHY 1261 00:43:51,600 --> 00:43:52,680 CONTROLS WERE SIGNIFICANT FOR 1262 00:43:52,680 --> 00:43:54,640 ALL THREE CELL TYPES. 1263 00:43:54,640 --> 00:43:56,440 PAIR WISE COMPARISONS BETWEEN 1264 00:43:56,440 --> 00:43:58,440 MODERATE RISK PA PATIENTS AND 1265 00:43:58,440 --> 00:43:59,240 HEALTHY CONTROLS WERE 1266 00:43:59,240 --> 00:44:00,960 SIGNIFICANT FOR THE NEUTROPHILS 1267 00:44:00,960 --> 00:44:04,880 AND THE MONOCYTES AND FINALLY 1268 00:44:04,880 --> 00:44:06,840 PAIR WISE COMPARISONS BETWEEN 1269 00:44:06,840 --> 00:44:09,160 HIGH RISK PA PATIENTS AND 1270 00:44:09,160 --> 00:44:10,160 HEALTHY CONTROLS WERE 1271 00:44:10,160 --> 00:44:11,400 SIGNIFICANT FOR ALL THREE CELL 1272 00:44:11,400 --> 00:44:11,800 TYPES. 1273 00:44:11,800 --> 00:44:14,400 IN ALL INSTANCES CELL FREE DNA 1274 00:44:14,400 --> 00:44:15,480 FROM INFLAMMATORY CELLS WERE 1275 00:44:15,480 --> 00:44:17,960 HIGHER IN PH PATIENTS. 1276 00:44:17,960 --> 00:44:20,600 I WOULD NOW LIKE TO BRIEFLY 1277 00:44:20,600 --> 00:44:24,040 DISCUSS THE BASIS FOR OUR 1278 00:44:24,040 --> 00:44:26,040 SPURONO LACTONE INTERVENTIONAL 1279 00:44:26,040 --> 00:44:27,000 PULMONARY HYPERTENSION TRIAL. 1280 00:44:27,000 --> 00:44:28,800 CURRENTLY NONE OF THE IMPROVE 1281 00:44:28,800 --> 00:44:30,640 TREATMENT FOR PAH DIRECTLY 1282 00:44:30,640 --> 00:44:32,200 LIMITS INFLAMMATION. 1283 00:44:32,200 --> 00:44:33,880 DR. ELINOFF HAS ALREADY REVIEWED 1284 00:44:33,880 --> 00:44:37,080 THE MOLECULAR DATA SUGGESTING 1285 00:44:37,080 --> 00:44:39,200 SPIRONO LACTONE HAS 1286 00:44:39,200 --> 00:44:40,760 ANTI-INFLAMMATORY EFFECTS IN 1287 00:44:40,760 --> 00:44:41,920 PULMONARY ARTERIAL HYPERTENSION. 1288 00:44:41,920 --> 00:44:44,120 WE ALSO PERFORM A RETROSPECTIVE 1289 00:44:44,120 --> 00:44:45,800 COHORT ANALYSIS TO INVESTIGATE 1290 00:44:45,800 --> 00:44:48,480 THE EFFECTS OF SPIRONO LACTONE 1291 00:44:48,480 --> 00:44:50,240 THERAPY ON CIRCULATING MEDIATORS 1292 00:44:50,240 --> 00:44:52,520 OF INFLAMMATION. 1293 00:44:52,520 --> 00:44:55,280 ON THE LEFT THE INDICATOR SHOWS 1294 00:44:55,280 --> 00:44:57,840 DECREASES IN SERUM MEDIATED 1295 00:44:57,840 --> 00:44:59,200 CONCENTRATIONS SHOWN IN BLUE, 1296 00:44:59,200 --> 00:45:01,560 AND INCREASES IN ORANGE. 1297 00:45:01,560 --> 00:45:04,320 AS YOU CAN SEE, TREATMENT WITH 1298 00:45:04,320 --> 00:45:06,640 SPIRONO LACTONE WAS CONSISTENTLY 1299 00:45:06,640 --> 00:45:08,680 ASSOCIATED WITH DECREASED 1300 00:45:08,680 --> 00:45:10,680 CONCENTRATIONS OF INFLAMMATORY 1301 00:45:10,680 --> 00:45:11,960 MEDIATORS WHEN COMPARED TO 1302 00:45:11,960 --> 00:45:14,080 PATIENTS WHO WERE NOT TREATED 1303 00:45:14,080 --> 00:45:16,320 WITH SPIRONO LACTONE. 1304 00:45:16,320 --> 00:45:20,200 IN STARK CONTRAST, TREATMENT BE 1305 00:45:20,200 --> 00:45:23,520 ENDOTHELIN 1 RECEPTOR 1306 00:45:23,520 --> 00:45:26,160 ANTAGONIST, A PH SPECIFIC 1307 00:45:26,160 --> 00:45:28,720 THERAPY WAS INCREASED WITH 1308 00:45:28,720 --> 00:45:30,080 ANTI-INFLAMMATORY CONCENTRATIONS 1309 00:45:30,080 --> 00:45:31,080 AND OTHER DRUGS WERE ASSOCIATED 1310 00:45:31,080 --> 00:45:32,720 WITH MIXED EFFECTS. 1311 00:45:32,720 --> 00:45:35,600 TO DATE, WE HAVE ENROLLED 37 OF 1312 00:45:35,600 --> 00:45:39,040 A PLANNED 50 PATIENTS IN OUR 1313 00:45:39,040 --> 00:45:42,120 SPIRONO LACTONE RANDOMIZED 1314 00:45:42,120 --> 00:45:42,760 INTERVENTIONAL TRIAL. 1315 00:45:42,760 --> 00:45:45,200 THE STUDY IS RANDOMIZE DOUBLE 1316 00:45:45,200 --> 00:45:46,600 BLINDED PLACEBO CONTROL PHASE 1, 1317 00:45:46,600 --> 00:45:49,120 2 TRIAL, PRIMARY END POINTS 1318 00:45:49,120 --> 00:45:50,520 INCLUDES CHANGE IN PLACEBO 1319 00:45:50,520 --> 00:45:51,720 CORRECTED SIX MINUTE WALK 1320 00:45:51,720 --> 00:45:53,280 DISTANCE AND INCIDENCE OF 1321 00:45:53,280 --> 00:45:55,120 CLINICAL WORSENING. 1322 00:45:55,120 --> 00:45:56,880 THIS SLIDE ILLUSTRATES THE TIME 1323 00:45:56,880 --> 00:46:00,240 LINE OF OUR SPIR PH TRIAL, 1324 00:46:00,240 --> 00:46:02,480 PATIENTS UNDERGO BASELINE 1325 00:46:02,480 --> 00:46:04,280 TESTING CONSISTING OF CLINICAL 1326 00:46:04,280 --> 00:46:05,440 ASSESS. 1327 00:46:05,440 --> 00:46:07,600 , SIX MINUTE WALK TESTING, ECHO 1328 00:46:07,600 --> 00:46:09,400 CARD YES GRAM AND CARDIAC MRI 1329 00:46:09,400 --> 00:46:10,640 AND BLOOD RESEARCH TESTING. 1330 00:46:10,640 --> 00:46:12,320 PATIENTS WERE RANDOMIZED TO 1331 00:46:12,320 --> 00:46:14,240 STUDY LAB OR PLACEBO, LAB CHECKS 1332 00:46:14,240 --> 00:46:16,800 ARE DONE AND IF ALL WELL 1333 00:46:16,800 --> 00:46:18,480 PATIENTS DUCK YOU KNOW DOUBLED. 1334 00:46:18,480 --> 00:46:20,920 THEY RETURN FOR REPEAT TESTING 1335 00:46:20,920 --> 00:46:22,280 AT 12 AND 24 WEEKS. 1336 00:46:22,280 --> 00:46:23,240 PATIENTS CONTINUE RESEARCH 1337 00:46:23,240 --> 00:46:24,680 PARTICIPATION AT THE NIH 1338 00:46:24,680 --> 00:46:26,000 CLINICAL CENTER THROUGH NATURAL 1339 00:46:26,000 --> 00:46:28,280 HISTORY PROTOCOL TO DATE WE HAVE 1340 00:46:28,280 --> 00:46:31,520 ENROLLED 86 PATIENTS IN OUR 1341 00:46:31,520 --> 00:46:34,720 NATURAL HISTORY PULMONARY 1342 00:46:34,720 --> 00:46:36,400 ARTERIAL HYPERTENSION PROTOCOL. 1343 00:46:36,400 --> 00:46:38,360 KEY POINTS NORMAL RESPONSE 1344 00:46:38,360 --> 00:46:39,800 DETECTED IN BLOOD OF PH PATIENTS 1345 00:46:39,800 --> 00:46:41,160 ELEVATED CELL FREE DNA LEVELS 1346 00:46:41,160 --> 00:46:42,600 DETECTED IN PAH AND ASSOCIATED 1347 00:46:42,600 --> 00:46:44,480 WITH TISSUE INJURY PATTERN 1348 00:46:44,480 --> 00:46:45,800 INVOLVING INFLAMMATORY CELLS. 1349 00:46:45,800 --> 00:46:50,000 IN I A DIVISION TO CURRENT 1350 00:46:50,000 --> 00:46:52,680 PULMONARY VASODILATED THERAPY 1351 00:46:52,680 --> 00:46:54,920 TARGETED INFLAMMATION MAY REVEAL 1352 00:46:54,920 --> 00:46:55,880 A NEW TREATMENT PARADIGM. 1353 00:46:55,880 --> 00:46:57,920 DR. ELINOFF WILL NOW DESCRIBE 1354 00:46:57,920 --> 00:46:59,480 HOW WE HAVE TAKEN THE RESEARCH 1355 00:46:59,480 --> 00:47:01,120 BACK TO BENCH WITH PLANS TO 1356 00:47:01,120 --> 00:47:02,640 COLLABORATE WITH NCATS TO 1357 00:47:02,640 --> 00:47:04,200 LEVERAGE THEIR DRUG DISCOVERY 1358 00:47:04,200 --> 00:47:05,880 TOOLBOX TO PRODUCE SIMILAR 1359 00:47:05,880 --> 00:47:07,960 COMPOUNDS WITH ENHANCED BENEFIT 1360 00:47:07,960 --> 00:47:10,000 TO ADVERSE DRUG REACTION RATIOS. 1361 00:47:10,000 --> 00:47:11,880 I WILL TAKE THE BATON AND PASS 1362 00:47:11,880 --> 00:47:12,840 IT BACK TO HIM. 1363 00:47:12,840 --> 00:47:17,680 >> THANK YOU, MICHAEL. 1364 00:47:17,680 --> 00:47:23,560 SO IN THE LAST PART, OF OUR 1365 00:47:23,560 --> 00:47:25,560 PRESENTATION TODAY MOVING FROM 1366 00:47:25,560 --> 00:47:26,920 THE BEDSIDE BACK TO BENCH. 1367 00:47:26,920 --> 00:47:29,800 LY DISCUSS NEXT STEPS IN OUR 1368 00:47:29,800 --> 00:47:32,920 INVESTIGATION OF XPV DEGRADATION 1369 00:47:32,920 --> 00:47:36,080 AS NOVEL MECHANISM FOR TREATING 1370 00:47:36,080 --> 00:47:36,600 ENOPLIA FLAYMATION. 1371 00:47:36,600 --> 00:47:43,080 EARLY YE I PRODUCED SPIRONO 1372 00:47:43,080 --> 00:47:43,600 LACTONE. 1373 00:47:43,600 --> 00:47:45,720 THIS IS A DRUG DISCOVERY 1374 00:47:45,720 --> 00:47:46,600 PARADIGM AILING ATTORNEYTIVE TO 1375 00:47:46,600 --> 00:47:48,000 INHIBITING PROTEIN FUNCTION 1376 00:47:48,000 --> 00:47:49,640 THROUGH ACTIVE SITE BINDING. 1377 00:47:49,640 --> 00:47:50,840 THIS STRATEGY TARGETS WHAT IS 1378 00:47:50,840 --> 00:47:53,160 GENERALLY CONSIDERED THE 1379 00:47:53,160 --> 00:47:54,520 UNDRUGGABLE PROTEOME. 1380 00:47:54,520 --> 00:47:56,440 INCLUDING TRANSCRIPTION FACTORS, 1381 00:47:56,440 --> 00:47:59,920 SCAFFOLDING PROTEINS AND UNOTHER 1382 00:47:59,920 --> 00:48:00,960 NON-ENZYMATIC PROTEINS. 1383 00:48:00,960 --> 00:48:02,680 HERE ON THE LEFT IS SCHEMATIC OF 1384 00:48:02,680 --> 00:48:06,000 THE TF 2H TRANSCRIPTION FACTOR 1385 00:48:06,000 --> 00:48:07,000 COMPLEX. 1386 00:48:07,000 --> 00:48:08,400 FOLLOWING IN VITRO TREATMENT OF 1387 00:48:08,400 --> 00:48:13,200 CELLS WITH SPIRONO LACTONE THE 1388 00:48:13,200 --> 00:48:15,400 PROXIMATE KNOWN STEP IN XPV 1389 00:48:15,400 --> 00:48:19,040 DEGRADATION IS CDK 7 DEPENDENT 1390 00:48:19,040 --> 00:48:22,200 PHOSPHORYLATION OF XPV. 1391 00:48:22,200 --> 00:48:25,560 NEXT LEADS TO FORMATION OFCF 1392 00:48:25,560 --> 00:48:27,440 XPB UBIQUITINATION. 1393 00:48:27,440 --> 00:48:29,960 ONCE UBIQUITINATED VCP 1394 00:48:29,960 --> 00:48:31,440 SEGREGATES APPEARS REQUIRED FOR 1395 00:48:31,440 --> 00:48:34,520 SUBSEQUENT DEGRADATION OF THE 1396 00:48:34,520 --> 00:48:36,120 26S PROTEOSOME. 1397 00:48:36,120 --> 00:48:37,120 HOWEVER THERE ARE POTENTIAL 1398 00:48:37,120 --> 00:48:38,920 LIMITATIONS TO TRANSLATING THE 1399 00:48:38,920 --> 00:48:41,560 MECHANISM OF SPIRONO LACTONE 1400 00:48:41,560 --> 00:48:43,480 INDUCED DEGRADATION INTO 1401 00:48:43,480 --> 00:48:45,920 CLINICALLY RELEVANT OUTCOMES. 1402 00:48:45,920 --> 00:48:46,840 ON THE LEFT YOU CAN SEE 1403 00:48:46,840 --> 00:48:49,200 SIGNIFICANT REDUCTION OF XPV 1404 00:48:49,200 --> 00:48:52,480 BEGINS SPIRONO LACTONE 1405 00:48:52,480 --> 00:48:54,320 CONCENTRATIONS OF 3 MICROMOLAR 1406 00:48:54,320 --> 00:48:55,640 IN VITRO WHICH MAY NOT BE 1407 00:48:55,640 --> 00:48:56,040 ACHIEVABLE. 1408 00:48:56,040 --> 00:48:57,240 DATA FROM A RECENT TRIAL IN 1409 00:48:57,240 --> 00:48:58,840 PATIENTS WITH HEART FAILURE AND 1410 00:48:58,840 --> 00:49:01,200 PRESERVE EJECTION FRACTION, AT 1411 00:49:01,200 --> 00:49:05,120 DOSES UP TO 45 MILLIGRAMS THE 1412 00:49:05,120 --> 00:49:07,280 MEDIAN CONCENTRATION MAJOR 1413 00:49:07,280 --> 00:49:09,240 METABOLITE WAS WELL BELOW 1 1414 00:49:09,240 --> 00:49:09,800 MICROMOLAR. 1415 00:49:09,800 --> 00:49:11,840 THIS IS PARTICULARLY RELEVANT 1416 00:49:11,840 --> 00:49:15,240 GIVEN THAT SPIRONO LACTONE 1417 00:49:15,240 --> 00:49:16,920 ADVERSE EVENTS ARE DOSE 1418 00:49:16,920 --> 00:49:18,040 DEPENDENT AN LIMIT AMOUNT OF 1419 00:49:18,040 --> 00:49:20,640 DRUG SAFELY TOLERATED. 1420 00:49:20,640 --> 00:49:23,400 ADDITIONALLY, IN VIVO SPIRONO 1421 00:49:23,400 --> 00:49:24,920 LACTONE IS CONVERTED TO TWO 1422 00:49:24,920 --> 00:49:26,400 MAJOR METABOLITES. 1423 00:49:26,400 --> 00:49:30,000 7 ALPHA SPIRONO LACTONE AND -- 1424 00:49:30,000 --> 00:49:31,880 NOTED THE THE LAST SLIDE. 1425 00:49:31,880 --> 00:49:34,640 UNLIKE SHOWN IN GREEN, BENEATH 1426 00:49:34,640 --> 00:49:36,440 EVERY OF THESE METABOLITES 1427 00:49:36,440 --> 00:49:38,440 INDUCE XPV DEGRADATION. 1428 00:49:38,440 --> 00:49:41,400 LIKEWISE SHOWN EARLIER WHILE 1429 00:49:41,400 --> 00:49:43,200 SPIRONO LACTONE IN GREEN REDUCES 1430 00:49:43,200 --> 00:49:45,120 NF KAPPA B AN AP 1 REPORTER 1431 00:49:45,120 --> 00:49:46,640 ACTIVITY, THIS ANTI-INFLAMMATORY 1432 00:49:46,640 --> 00:49:47,800 I WASOR EFFECT IS NOT OBSERVED 1433 00:49:47,800 --> 00:49:49,120 WITH EITHER TWO MAJOR 1434 00:49:49,120 --> 00:49:50,840 METABOLITES. 1435 00:49:50,840 --> 00:49:52,320 THEREFORE, IN PARTNERSHIP WITH 1436 00:49:52,320 --> 00:49:53,720 THE EARLY TRANSLATION BRANCH AT 1437 00:49:53,720 --> 00:49:55,120 THE NATIONAL CENTER FOR 1438 00:49:55,120 --> 00:49:57,080 ADVANCING TRANSLATIONAL 1439 00:49:57,080 --> 00:49:58,760 SCIENCES, OR NCATS, OUR GOAL IS 1440 00:49:58,760 --> 00:50:01,000 TO IDENTIFY NEW THERAPEUTICALLY 1441 00:50:01,000 --> 00:50:03,000 RELEVANT COMPOUNDS THAT INDUCE 1442 00:50:03,000 --> 00:50:05,360 XPV DEGRADATION AND SUPPRESS 1443 00:50:05,360 --> 00:50:06,000 INFLAMMATION. 1444 00:50:06,000 --> 00:50:08,040 WE ALSO PROPOSE TO USE GENOME 1445 00:50:08,040 --> 00:50:10,000 WIDE RNAi SCREENING AS WELL AS 1446 00:50:10,000 --> 00:50:11,680 MOLECULAR MODELING AND STRUCTURE 1447 00:50:11,680 --> 00:50:14,080 BASED PREDICTION TOOLS IN ORDER 1448 00:50:14,080 --> 00:50:15,200 TO DETERMINE SPECIFIC BINDING 1449 00:50:15,200 --> 00:50:16,240 TARGETS. 1450 00:50:16,240 --> 00:50:17,920 THERE BY ADVANCING THE CURRENT 1451 00:50:17,920 --> 00:50:20,720 WORKING MODEL OF DRUG INDUCED 1452 00:50:20,720 --> 00:50:21,880 XPV DEGRADATION. 1453 00:50:21,880 --> 00:50:23,400 THE FIRST STEP IN THIS PROJECT 1454 00:50:23,400 --> 00:50:25,960 WAS TO DESIGN AND VALIDATE ASSAY 1455 00:50:25,960 --> 00:50:28,240 SYSTEM THAT WAS AMENABLE TO HIGH 1456 00:50:28,240 --> 00:50:29,400 THROUGH PUT SCREENING. 1457 00:50:29,400 --> 00:50:31,600 IN COLLABORATION WITH KEN CHANG 1458 00:50:31,600 --> 00:50:33,200 IN THE FUNCTIONAL GENOMICS LAB 1459 00:50:33,200 --> 00:50:39,840 AT NCATS, CRISPR C SARKS 9 WAS 1460 00:50:39,840 --> 00:50:42,200 USED TO TAG PROTEIN WITH 1 # 1 1461 00:50:42,200 --> 00:50:44,520 AMINO ACID LUMINESCENT REPORTER 1462 00:50:44,520 --> 00:50:46,080 PEPTIDE IN HUMAN EMBRYONIC 1463 00:50:46,080 --> 00:50:46,680 KIDNEY CELLS. 1464 00:50:46,680 --> 00:50:50,320 DUE TO ITS SMALL SIZE, IT CAN BE 1465 00:50:50,320 --> 00:50:52,000 EFFECTIVELY INSERTED TO THE 1466 00:50:52,000 --> 00:50:52,920 GENOME WITHOUT DELETERIOUS 1467 00:50:52,920 --> 00:50:54,360 EFFECTS TO THE FUNCTION OR 1468 00:50:54,360 --> 00:50:55,800 LOCALIZATION OF THE TAG 1469 00:50:55,800 --> 00:50:56,960 ENDOGENOUS PROTEIN. 1470 00:50:56,960 --> 00:50:59,760 IN OUR CASE, FOUR GUIDE RNAs 1471 00:50:59,760 --> 00:51:01,240 WERE TESTED FOR GENE EDITING. 1472 00:51:01,240 --> 00:51:03,120 THIS RESULTED IN FOUR DIFFERENT 1473 00:51:03,120 --> 00:51:06,840 CELL LINES EXPRESSING XPV HIGH 1474 00:51:06,840 --> 00:51:08,120 BIT FUSION PROTEIN. 1475 00:51:08,120 --> 00:51:10,880 WHEN CELLS CONTAINING HIGH BIT 1476 00:51:10,880 --> 00:51:14,640 TAG XPV ARE LYSED AND COMPLEX 1477 00:51:14,640 --> 00:51:18,840 WITH L G BIT THEY RECONSTITUTE A 1478 00:51:18,840 --> 00:51:19,480 SENSITIVE LYM LESS SENT REPORTER 1479 00:51:19,480 --> 00:51:20,720 AND LIGHT IS REDUCED IN 1480 00:51:20,720 --> 00:51:23,120 PROPORTION TO PROTEIN LEVELS. 1481 00:51:23,120 --> 00:51:25,880 T SHOWN ON THE LEFT THE Y AXIS 1482 00:51:25,880 --> 00:51:29,760 REPRESENTS FULL CHANGE IN 1483 00:51:29,760 --> 00:51:31,680 LUMINESCENCE OF THE TRANSFECTED 1484 00:51:31,680 --> 00:51:32,160 CELLS. 1485 00:51:32,160 --> 00:51:35,040 OF THE FOUR RNAs, 3 AND 4 HAD 1486 00:51:35,040 --> 00:51:36,360 HIGHEST INCREASE IN 1487 00:51:36,360 --> 00:51:37,640 LUMINESCENCE. 1488 00:51:37,640 --> 00:51:40,280 WE EXAMINED SPIRONO LACTONE 1489 00:51:40,280 --> 00:51:41,400 INDUCED DEGRADATION. 1490 00:51:41,400 --> 00:51:43,000 ON THE RIGHT WESTERN BLOTTING 1491 00:51:43,000 --> 00:51:45,760 WITH AN ANTI-XPV ANTIBODY SHOWS 1492 00:51:45,760 --> 00:51:47,400 REDUCTION OF XPV IN THE PRESENCE 1493 00:51:47,400 --> 00:51:49,600 OF SPIRON LACTONE BUT MORE 1494 00:51:49,600 --> 00:51:51,280 IMPORTANTLY USING ANTI-HIGH BIT 1495 00:51:51,280 --> 00:51:53,720 ANTIBODY WE SEE THAT THE X PV 1496 00:51:53,720 --> 00:51:55,280 HIGH BIT FUSION PROTEIN IS 1497 00:51:55,280 --> 00:51:56,720 SIMILARLY REDUCED IN THE 1498 00:51:56,720 --> 00:51:59,240 PRESENCE OF SPIRONO LACTONE 1499 00:51:59,240 --> 00:52:02,120 DEMONSTRATING IT REMAINS 1500 00:52:02,120 --> 00:52:03,240 SUSCEPTIBLE TO INDUCED 1501 00:52:03,240 --> 00:52:04,720 DEGRADATION. 1502 00:52:04,720 --> 00:52:07,160 CONSISTENT WITH SPIRONO LACTONE 1503 00:52:07,160 --> 00:52:09,240 INDUCED DEGRADATION OF HIGH BIT 1504 00:52:09,240 --> 00:52:11,520 FUSION PROTEIN WE SEE REDUCTION 1505 00:52:11,520 --> 00:52:13,400 IN LUMINESCENCE FOLLOWING 1506 00:52:13,400 --> 00:52:16,240 TREATMENT FOR ONE, TWO AND 4 1507 00:52:16,240 --> 00:52:17,000 FORCE. 1508 00:52:17,000 --> 00:52:18,560 THUS THIS ASSAY SYSTEM CAN BE 1509 00:52:18,560 --> 00:52:20,600 USED FOR HIGH THROUGH PUT SMALL 1510 00:52:20,600 --> 00:52:22,480 MOLECULE SCREENING TO IDENTIFY 1511 00:52:22,480 --> 00:52:25,160 OTHER COMPOUNDS THAT INDUCE XPV 1512 00:52:25,160 --> 00:52:26,800 DEGRADATION AS A WELL AS A 1513 00:52:26,800 --> 00:52:28,520 PLATFORM FOR GENOME WIDE RNAi 1514 00:52:28,520 --> 00:52:29,720 SCREENING. 1515 00:52:29,720 --> 00:52:31,320 IN ADDITION TO CONFIRMING THAT 1516 00:52:31,320 --> 00:52:33,040 THE FUSION PROTEIN IS 1517 00:52:33,040 --> 00:52:35,920 SUSCEPTIBLE TO SPIRONO LACTONE 1518 00:52:35,920 --> 00:52:38,000 INDUCED DEGRADATION IT WAS USED 1519 00:52:38,000 --> 00:52:39,360 TO VERIFY LOCALIZATION AND 1520 00:52:39,360 --> 00:52:42,120 FUNCTION OF THE HIGH BIT TAG 1521 00:52:42,120 --> 00:52:43,480 XPB: AS SHOWN HERE STAINED IN 1522 00:52:43,480 --> 00:52:46,880 RED, HIGH BIT TAG XPB APPEARS 1523 00:52:46,880 --> 00:52:48,480 PREDOMINANTLY LOCALIZED TO THE 1524 00:52:48,480 --> 00:52:50,000 NUCLEUS. 1525 00:52:50,000 --> 00:52:51,320 SIMILAR TO THE BLUE NUCLEAR 1526 00:52:51,320 --> 00:52:51,520 STAIN. 1527 00:52:51,520 --> 00:52:52,960 THIS IS HIGHLIGHTED IN THE 1528 00:52:52,960 --> 00:52:54,840 MERGED IMAGE TO THE RIGHT WHERE 1529 00:52:54,840 --> 00:52:56,440 OVERLAPPING OF RED AND BLUE 1530 00:52:56,440 --> 00:52:59,560 RESULT IN A PURPLE COLOR. 1531 00:52:59,560 --> 00:53:01,040 FURTHERMORE WHEN CELLS TREAT 1532 00:53:01,040 --> 00:53:04,040 WITH SPIRONO LACTONE SIGNIFICANT 1533 00:53:04,040 --> 00:53:05,720 REDUCTION IN RED STAINING HIGH 1534 00:53:05,720 --> 00:53:08,560 BIT TAG XPB LEADS BLUE NUCLEAR 1535 00:53:08,560 --> 00:53:10,600 STAINENING THE MERGED IMAGE -- 1536 00:53:10,600 --> 00:53:11,640 STAINENING THE MERGED IMAGE TO 1537 00:53:11,640 --> 00:53:13,000 THE RIGHT. 1538 00:53:13,000 --> 00:53:14,480 IN TERMS OF FUNCTION IN PRESENCE 1539 00:53:14,480 --> 00:53:17,040 OF INFLAMMATORY STIMULUS, THE NF 1540 00:53:17,040 --> 00:53:18,960 KAPPA B AN AP 1 REPORTER GENES 1541 00:53:18,960 --> 00:53:22,040 ARE ACTIVATED IN HEP 293 CELLS 1542 00:53:22,040 --> 00:53:23,040 EXPRESSING THE HIGH BIT FUSION 1543 00:53:23,040 --> 00:53:23,880 PROTEIN. 1544 00:53:23,880 --> 00:53:26,040 AND AS EXPECTED, GENE ACTIVATION 1545 00:53:26,040 --> 00:53:28,080 IS SUPPRESSED BY SPIRONO 1546 00:53:28,080 --> 00:53:28,680 LACTONE. 1547 00:53:28,680 --> 00:53:30,120 NOW WITH A VALIDATED WORKING 1548 00:53:30,120 --> 00:53:31,880 HIGH THROUGH PUT ASSAY IN HAND 1549 00:53:31,880 --> 00:53:34,200 WE PLAN TO PROCEED WITH 1550 00:53:34,200 --> 00:53:36,680 SCREENING NCATS SMALL MOLECULE 1551 00:53:36,680 --> 00:53:38,120 LIBRARIES INCLUDING PHARMACO 1552 00:53:38,120 --> 00:53:40,640 LOGICALLY ANNOTATED CHEMICAL 1553 00:53:40,640 --> 00:53:41,800 TOOLBOX, COMPLIMENTARY 1554 00:53:41,800 --> 00:53:43,360 APPROACHES WOULD INCLUDE A 1555 00:53:43,360 --> 00:53:45,040 GENOME WIDE RNAi SCREEN 1556 00:53:45,040 --> 00:53:46,520 MOLECULAR MODELING AND STRUCTURE 1557 00:53:46,520 --> 00:53:48,800 BASED PREDICTION TOOLS. 1558 00:53:48,800 --> 00:53:50,320 COMPOUNDS IDENTIFIED IN THIS 1559 00:53:50,320 --> 00:53:51,440 INITIAL SCREEN WOULD BE TESTED 1560 00:53:51,440 --> 00:53:54,120 FOR ANTI-INFLAMMATORY ACTIVITY 1561 00:53:54,120 --> 00:53:56,720 AS WELL AS UNDERGO ADDITIONAL 1562 00:53:56,720 --> 00:53:58,400 MECHANISTIC STUDIES. 1563 00:53:58,400 --> 00:54:00,000 LEAD COMPOUNDS WOULD BE TESTED 1564 00:54:00,000 --> 00:54:02,080 IN OUR HUMAN ENDOTHELIAL CELL 1565 00:54:02,080 --> 00:54:04,680 MODELS OF PAH AND UNDERGO 1566 00:54:04,680 --> 00:54:05,800 PHARMACOKINETIC AND 1567 00:54:05,800 --> 00:54:07,360 PHARMACODYNAMIC OPTIMIZATION. 1568 00:54:07,360 --> 00:54:09,160 FOLLOWED BY PRE-CLINICAL TESTING 1569 00:54:09,160 --> 00:54:12,520 IN PAH SMALL ANIMAL MODELS AND 1570 00:54:12,520 --> 00:54:13,520 ULTIMATELY EARLY PHASE CLINICAL 1571 00:54:13,520 --> 00:54:15,240 TRIALS CONDUCTED HERE AT THE NIH 1572 00:54:15,240 --> 00:54:17,520 CLINICAL CENTER. 1573 00:54:17,520 --> 00:54:19,160 CRITICAL TO THIS ENDEAVOR WE 1574 00:54:19,160 --> 00:54:20,960 ESTABLISHED A RELEVANT RAT MODEL 1575 00:54:20,960 --> 00:54:22,520 PAH IN ORDER TO INVESTIGATE 1576 00:54:22,520 --> 00:54:23,880 PROMISING THERAPEUTIC TARGETS 1577 00:54:23,880 --> 00:54:25,200 IDENTIFIED IN OUR LAB. 1578 00:54:25,200 --> 00:54:27,000 RATS ARE INJECTED SUBCUTANEOUSLY 1579 00:54:27,000 --> 00:54:30,800 WITH A SINGLE DOSE OF SU 5416 1580 00:54:30,800 --> 00:54:32,160 TYROSINE KINASE INHIBITOR. 1581 00:54:32,160 --> 00:54:35,160 THEN EXPOSED TO HYPOXIA FOR 1582 00:54:35,160 --> 00:54:35,800 THREE WEEKS. 1583 00:54:35,800 --> 00:54:39,000 AFTER THEY RETURN TO NORM OXIA 1584 00:54:39,000 --> 00:54:40,840 FOR THE REMAINDER OF THE STUDY. 1585 00:54:40,840 --> 00:54:44,280 IN THIS THE RATS DEVELOPED 1586 00:54:44,280 --> 00:54:45,600 SEVERE DYSFUNCTION AS SHOWN BY 1587 00:54:45,600 --> 00:54:47,280 CARDIAC MRI WHERE THE RATIO OF 1588 00:54:47,280 --> 00:54:50,080 THE THE RV AND DIASTOLIC VOLUME 1589 00:54:50,080 --> 00:54:53,480 TO LV DIASTOLIC VOLUME INCREASE. 1590 00:54:53,480 --> 00:54:57,480 IN CARDIAC INDEX IS REDUCED THE 1591 00:54:57,480 --> 00:54:58,400 PA CLASS. 1592 00:54:58,400 --> 00:55:00,000 INVASIVE HEMODYNAMIC 1593 00:55:00,000 --> 00:55:01,160 MEASUREMENTS AT TEN WEEKS 1594 00:55:01,160 --> 00:55:03,240 DEMONSTRATE SIGNIFICANTLY 1595 00:55:03,240 --> 00:55:04,920 ELEVATED PULMONARY ARTERY 1596 00:55:04,920 --> 00:55:07,240 PRESSURE, AND LUNG HISTOLOGY 1597 00:55:07,240 --> 00:55:08,640 DEMONSTRATES SEVERE VASCULAR 1598 00:55:08,640 --> 00:55:10,120 REMODELING THAT CLOSELY 1599 00:55:10,120 --> 00:55:11,480 RESEMBLES THE CHANGES FOUND IN 1600 00:55:11,480 --> 00:55:13,520 LUNGS FOR PAH PATIENTS. 1601 00:55:13,520 --> 00:55:16,360 IN SUMMARY, INDUCING XPV 1602 00:55:16,360 --> 00:55:17,760 DEGRADATION IS A NOVEL STRATEGY 1603 00:55:17,760 --> 00:55:19,320 TORR TREATING VASCULAR 1604 00:55:19,320 --> 00:55:21,280 INFLAMMATION IN PAH THAT MAY BE 1605 00:55:21,280 --> 00:55:23,720 APPLICABLE TO A WIDE RANGE OF 1606 00:55:23,720 --> 00:55:25,600 CARDIOVASCULAR CONDITIONS. 1607 00:55:25,600 --> 00:55:26,560 ANTI-INFLAMMATORY COMPOUNDS WITH 1608 00:55:26,560 --> 00:55:29,400 UNIQUE MECHANISM OF ACTION IDE 1609 00:55:29,400 --> 00:55:31,640 ANIED MAY OFFER NEW OPTIONS FOR 1610 00:55:31,640 --> 00:55:32,880 PATIENTS SUFFERING FROM 1611 00:55:32,880 --> 00:55:38,800 AUTOIMMUNE OR AUTOI BELIEVE AUTY 1612 00:55:38,800 --> 00:55:39,040 DISEASE. 1613 00:55:39,040 --> 00:55:41,240 THE SELECTION OF PROTEIN 1614 00:55:41,240 --> 00:55:42,360 DEGRADATION HAS BROAD 1615 00:55:42,360 --> 00:55:44,360 IMPLICATIONS FOR DRUG DISCOVERY 1616 00:55:44,360 --> 00:55:47,680 AND DEVELOPMENT AND MAY ENABLE 1617 00:55:47,680 --> 00:55:49,920 TARGETING MOLECULES HEAVE 1618 00:55:49,920 --> 00:55:50,720 EVIDENCE PREVIOUSLY THOUGHT 1619 00:55:50,720 --> 00:55:51,320 UNDRUGGABLE. 1620 00:55:51,320 --> 00:55:53,200 IN CONCLUSION WE HOPE WE 1621 00:55:53,200 --> 00:55:55,320 EMPHASIZED THE CURRENT NEED FOR 1622 00:55:55,320 --> 00:55:58,360 THERAPIES IN PAH GIVEN THAT OUR 1623 00:55:58,360 --> 00:56:00,560 CURRENT ARMAMENTARIUM DOESN'T 1624 00:56:00,560 --> 00:56:02,120 OFFER A CURE AND RATHER THAN 1625 00:56:02,120 --> 00:56:04,200 ONLY TREATING ELEVATED PULMONARY 1626 00:56:04,200 --> 00:56:06,480 ARTERIAL PRESSURE, IN RIGHT 1627 00:56:06,480 --> 00:56:07,720 VENTRICULAR DYSFUNCTION AT TIP 1628 00:56:07,720 --> 00:56:09,160 OF THE ICEBERG WE NEED TO LOOK 1629 00:56:09,160 --> 00:56:11,440 DEEPER TO APPRECIATE THE COMPLEX 1630 00:56:11,440 --> 00:56:13,440 INTERRELATED MOLECULAR 1631 00:56:13,440 --> 00:56:13,840 PROCESSES. 1632 00:56:13,840 --> 00:56:17,880 IN PAH PULMONARY VASCULAR 1633 00:56:17,880 --> 00:56:19,160 LESIONS SUBSIST IN A 1634 00:56:19,160 --> 00:56:20,120 MICROENVIRONMENT UNTARGETTED BY 1635 00:56:20,120 --> 00:56:22,400 CURRENT THERAPIST. 1636 00:56:22,400 --> 00:56:22,760 -- THERAPIES. 1637 00:56:22,760 --> 00:56:24,120 OUR PROGRAM IS LEVERAGING A 1638 00:56:24,120 --> 00:56:25,120 BENCH TO BEDSIDE AND BACK 1639 00:56:25,120 --> 00:56:27,360 APPROACH TO IDENTIFY NOVEL 1640 00:56:27,360 --> 00:56:28,400 THERAPIES THAT CAN BE TESTED 1641 00:56:28,400 --> 00:56:31,040 HERE AT THE NIH CLINICAL CENTER 1642 00:56:31,040 --> 00:56:32,960 WITH THE HOPE ONE DAY WE WILL BE 1643 00:56:32,960 --> 00:56:34,000 ABLE TO OFFER OUR PATIENTS A 1644 00:56:34,000 --> 00:56:36,200 CURE. 1645 00:56:36,200 --> 00:56:38,520 ON BEHALF OF MIKE AND I WE WOULD 1646 00:56:38,520 --> 00:56:40,560 LIKE TO THANK THE ENTIRE PAH 1647 00:56:40,560 --> 00:56:42,440 GROUP HERE IN THE CRITICAL CARE 1648 00:56:42,440 --> 00:56:44,640 MEDICINE DEPARTMENT, WE COULDN'T 1649 00:56:44,640 --> 00:56:46,120 HAVE DONE IT WITH SUCH A 1650 00:56:46,120 --> 00:56:48,120 WONDERFUL GROUP OF HARDWORKING 1651 00:56:48,120 --> 00:56:50,600 ENERGETIC BRILLIANT PEOPLE, ALSO 1652 00:56:50,600 --> 00:56:53,080 VERY THANKFUL FOR THE INTRAMURAL 1653 00:56:53,080 --> 00:56:55,280 COMMUNITY AND THE COLLABORATIONS 1654 00:56:55,280 --> 00:56:57,840 THAT WE HAVE HAD THAT HELPED 1655 00:56:57,840 --> 00:57:01,440 THESE PROJECTS AS WELL AS 1656 00:57:01,440 --> 00:57:04,960 EXTRAMURAL CLAY B COLLABORATOR. 1657 00:57:04,960 --> 00:57:07,280 WITH THAT WE ARE HAPPY TO TAKE 1658 00:57:07,280 --> 00:57:08,040 QUESTIONS WITH THE REMAINING 1659 00:57:08,040 --> 00:57:09,720 TIME. 1660 00:57:09,720 --> 00:57:12,000 >> THANK YOU, DR. SOLOMON AND 1661 00:57:12,000 --> 00:57:16,280 DR. ELINOFF FOR THE REALLY 1662 00:57:16,280 --> 00:57:17,640 WONDERFUL PRESENTATION ON THE 1663 00:57:17,640 --> 00:57:19,520 PATHOPHYSIOLOGY OF PAH AND ITS 1664 00:57:19,520 --> 00:57:24,040 GROUP THERAPIES AND YOUR WORK ON 1665 00:57:24,040 --> 00:57:26,240 SUSTAINING INFORMATION IN PAH 1666 00:57:26,240 --> 00:57:28,440 ESPECIALLY WITH YOUR SPIR TRIAL, 1667 00:57:28,440 --> 00:57:30,520 INSIGHTS IN YOUR COLLABORATION 1668 00:57:30,520 --> 00:57:32,760 ONE ANOTHER AND APPROACH OF 1669 00:57:32,760 --> 00:57:35,640 NCATS OFFERS SOME VERY EXCITING 1670 00:57:35,640 --> 00:57:37,000 THERAPEUTIC STRATEGIES FOR 1671 00:57:37,000 --> 00:57:39,640 DISEASE THAT IS HISTORICALLY 1672 00:57:39,640 --> 00:57:40,320 FRUSTRATINGLY DIFFICULT TO 1673 00:57:40,320 --> 00:57:41,200 TREAT. 1674 00:57:41,200 --> 00:57:43,520 BEFORE I GO ON TO QUESTIONS, I 1675 00:57:43,520 --> 00:57:45,880 WANT TO JUST FOR THE VIEWING 1676 00:57:45,880 --> 00:57:48,000 AUDIENCE, THE CORRECT CME CODE 1677 00:57:48,000 --> 00:57:52,120 TODAY WE GAVE YOU THE WRONG ONE 1678 00:57:52,120 --> 00:57:54,160 IS 36726. 1679 00:57:54,160 --> 00:57:55,760 PLEASE TEXT THE CODES TO THE 1680 00:57:55,760 --> 00:57:58,760 JOHNS HOPKINS CME PHONE NUMBER. 1681 00:57:58,760 --> 00:57:59,480 WE APOLOGIZE FOR THAT. 1682 00:57:59,480 --> 00:58:02,360 WE HAVE TIME FOR COUPLE OF 1683 00:58:02,360 --> 00:58:04,440 QUESTIONS. 1684 00:58:04,440 --> 00:58:07,480 ONE GIVEN THE EFFECT OF 1685 00:58:07,480 --> 00:58:09,040 (INDISCERNIBLE) IN JAK STAT 1686 00:58:09,040 --> 00:58:11,800 MEDIATED INFLAMMATION, IS THERE 1687 00:58:11,800 --> 00:58:17,160 A ROLE FOR NK 6 G2 CELLS IN PAH? 1688 00:58:17,160 --> 00:58:18,680 >> 1689 00:58:18,680 --> 00:58:22,760 >> THAT IS INTERESTING QUESTION. 1690 00:58:22,760 --> 00:58:25,320 SO SOME OF THE JAK STAT 1691 00:58:25,320 --> 00:58:27,320 INHIBITORS HAVE BEEN TESTED IN 1692 00:58:27,320 --> 00:58:31,720 ANIMAL MODELS OF PAH, CHRONIC 1693 00:58:31,720 --> 00:58:34,760 HYPOXIA MODEL MONOCLONALLY MODEL 1694 00:58:34,760 --> 00:58:36,600 THAT IS CHARACTERIZED BY QUITE 1695 00:58:36,600 --> 00:58:39,720 BIT OF INFLAMMATORY RESPONSE. 1696 00:58:39,720 --> 00:58:43,360 THEY HAVE BEEN SUCCESSFUL THERE. 1697 00:58:43,360 --> 00:58:46,000 AS FAR AS THAT EFFICACY BEING 1698 00:58:46,000 --> 00:58:50,760 ATTRIBUTED SPECIFICALLY TO NK 1699 00:58:50,760 --> 00:58:52,600 CELLS ACTUALLY THEIR WORK ON NK 1700 00:58:52,600 --> 00:58:55,160 CELLS WHICH IS A LITTLE OUTSIDE 1701 00:58:55,160 --> 00:58:58,600 MY EXPERTISE, HAS SHOWN IN THE 1702 00:58:58,600 --> 00:59:03,680 FIELD THERE BEING IMBALANCE NK 1703 00:59:03,680 --> 00:59:05,680 CELL SUB SETS IN PAH PATIENTS 1704 00:59:05,680 --> 00:59:11,440 AND THEN IN MODELS OF NK CELL 1705 00:59:11,440 --> 00:59:13,520 DEFICIENCY AND KNOCK OUT MOUSE 1706 00:59:13,520 --> 00:59:16,560 MORE SEVERE HYPOXIA RESPONSES IN 1707 00:59:16,560 --> 00:59:18,160 PH IN THAT ANIMAL MODEL. 1708 00:59:18,160 --> 00:59:20,760 SO THERE DOES APPEAR TO BE AN 1709 00:59:20,760 --> 00:59:24,520 IMPORTANT ROLE FOR NK CELLS IN 1710 00:59:24,520 --> 00:59:25,720 PH PATHOGENESIS. 1711 00:59:25,720 --> 00:59:29,720 OUR CELL FREE DNA ALSO DETECTED 1712 00:59:29,720 --> 00:59:33,720 THE CELL FREE DNA FROM THAT 1713 00:59:33,720 --> 00:59:36,720 LYMPHOCYTE SUBSET, WHETHER THE 1714 00:59:36,720 --> 00:59:38,080 EFFICACY OF JAK STAT INHIBITORS 1715 00:59:38,080 --> 00:59:41,080 ARE DUE TO SPECIFIC NK CELL, 1716 00:59:41,080 --> 00:59:45,880 SUBTYPE I'M NOT SURE YET. 1717 00:59:45,880 --> 00:59:46,320 >> OKAY. 1718 00:59:46,320 --> 00:59:48,200 THANK YOU. 1719 00:59:48,200 --> 00:59:52,000 FROM UNFORTUNATELY, THAT BRINGS 1720 00:59:52,000 --> 00:59:53,800 US UP TO -- I HAD A QUESTION 1721 00:59:53,800 --> 00:59:55,200 MYSELF I MAY EMAIL SEPARATELY. 1722 00:59:55,200 --> 00:59:56,800 THANK YOU, THAT BRINGS US UP TO 1723 00:59:56,800 --> 00:59:57,400 THE HOUR. 1724 00:59:57,400 --> 01:00:00,880 I WANT TO THANK BOTH OF YOU FOR 1725 01:00:00,880 --> 01:00:02,840 CAPTIVATING PRESENTATIONS AND WE 1726 01:00:02,840 --> 01:00:08,200 WISH ALL OF YOU MUCH SUCCESS IN 1727 01:00:08,200 --> 01:00:09,440 YOUR INVESTIGATIONS. 1728 01:00:09,440 --> 01:00:11,240 TO EVERYONE, HAVE A GREAT 1729 01:00:11,240 --> 01:00:11,480 AFTERNOON. 1730 01:00:11,480 --> 01:00:12,520 THANK YOU FOR JOINING US TODAY. 1731 01:00:12,520 --> 00:00:00,000 >>THANK YOU, EVERYONE.