1
00:00:05,720 --> 00:00:06,960
>>THANK YOU SO MUCH FOR
COMING

2
00:00:06,960 --> 00:00:07,360
OUT TODAY.

3
00:00:07,360 --> 00:00:09,240
THIS IS FANTASTIC TO SEE SO MANY

4
00:00:09,240 --> 00:00:11,720
PEOPLE HERE IN PERSON AND WE

5
00:00:11,720 --> 00:00:14,360
REALLY APPRECIATE THAT.

6
00:00:14,360 --> 00:00:17,200
I'M VERY HAPPY TO INTRODUCE

7
00:00:17,200 --> 00:00:17,720
SENTHIL MUTHUSWAMY TODAY.

8
00:00:17,720 --> 00:00:21,280
HE GOT HIS PH.D. FROM MCMASTER

9
00:00:21,280 --> 00:00:21,920
UNIVERSITY IN CANADA.

10
00:00:21,920 --> 00:00:24,360
HE DID A POSTDOCTORAL FELLOWSHIP

11
00:00:24,360 --> 00:00:27,040
WITH JOAN BRUGGE AND HE HAD

12
00:00:27,040 --> 00:00:28,160
SEVERAL FACULTY POSITIONS BEFORE

13
00:00:28,160 --> 00:00:29,600
COMING HERE, ONE COLDSPRING

14
00:00:29,600 --> 00:00:32,960
HARBOR, ANOTHER ONE, HE BECAME

15
00:00:32,960 --> 00:00:35,400
THE LEE K. AND MARGARET LAU

16
00:00:35,400 --> 00:00:37,960
CHAIR IN BREAST CANCER RESEARCH

17
00:00:37,960 --> 00:00:39,200
IN TORONTO, THEN HE BECAME THE

18
00:00:39,200 --> 00:00:40,520
DIRECTOR OF THE CELL BIOLOGY

19
00:00:40,520 --> 00:00:41,640
PROGRAM IN THE DEPARTMENT OF

20
00:00:41,640 --> 00:00:45,120
MEDICINE AT HARVARD MEDICAL.

21
00:00:45,120 --> 00:00:47,960
AND HE JOINED THE NCI IN 2022,

22
00:00:47,960 --> 00:00:51,360
WHERE HE BECAME MY LAB CHIEF AND

23
00:00:51,360 --> 00:00:52,800
MANY OTHER PEOPLE IN THE ROOM

24
00:00:52,800 --> 00:00:53,800
HERE TODAY.

25
00:00:53,800 --> 00:00:57,080
THE LABORATORY OF CANCER BIOLOGY

26
00:00:57,080 --> 00:00:57,960
AND GENETICS, AND WE'RE VERY

27
00:00:57,960 --> 00:01:01,520
HAPPY TO HAVE HIM HERE.

28
00:01:01,520 --> 00:01:02,840
HE'S RECEIVED NUMEROUS SCHOLAR

29
00:01:02,840 --> 00:01:04,720
AWARDS FOR HIS RESEARCH,

30
00:01:04,720 --> 00:01:08,800
INCLUDING ONE FROM MARIA ALLEN V

31
00:01:08,800 --> 00:01:09,400
FOUNDATION U.S. ARMY AIR OF

32
00:01:09,400 --> 00:01:12,440
HOPE, AND HE WAS AMONG THE FIRST

33
00:01:12,440 --> 00:01:15,120
TO ESTABLISH ORGANOID CULTURES

34
00:01:15,120 --> 00:01:17,640
TO EXPLORE 3D CELL-CELL

35
00:01:17,640 --> 00:01:18,920
INTERACTIONS AND THEIR

36
00:01:18,920 --> 00:01:19,920
TRANSLATIONAL POTENTIAL FOR

37
00:01:19,920 --> 00:01:21,560
UNDERSTANDING THE ROLE OF CELL

38
00:01:21,560 --> 00:01:23,480
POLARITY IN CANCER, THE ROLE OF

39
00:01:23,480 --> 00:01:26,240
DRUG RESISTANCE, AND IDENTIFYING

40
00:01:26,240 --> 00:01:28,760
NEW THERAPEUTIC MODALITIES.

41
00:01:28,760 --> 00:01:30,760
AND I HOPE IT OKAY TO SAY HE

42
00:01:30,760 --> 00:01:33,960
PLANS TO LAUNCH A CCR ORGANOID

43
00:01:33,960 --> 00:01:36,760
FACILITY WITH CATHY KELLY TO

44
00:01:36,760 --> 00:01:38,480
DEVELOP CLINICALLY A ANNOTATED

45
00:01:38,480 --> 00:01:39,320
ORGANOID LINES.

46
00:01:39,320 --> 00:01:42,920
SO TODAY, HE WILL SPEAK TO US ON

47
00:01:42,920 --> 00:01:44,840
ORGANOIDS AND CELL POLARITY IN

48
00:01:44,840 --> 00:01:46,120
TRANSLATIONAL AND MECHANISTIC

49
00:01:46,120 --> 00:01:47,200
CANCER RESEARCH.

50
00:01:47,200 --> 00:01:49,720
THANK YOU, SENTHIL.

51
00:01:49,720 --> 00:01:52,800
[APPLAUSE]

52
00:01:52,800 --> 00:01:55,320
THANK YOU VERY MUCH, BEV, FOR

53
00:01:55,320 --> 00:01:56,400
THAT NICE INTRODUCTION.

54
00:01:56,400 --> 00:01:57,560
IT'S ALWAYS A LITTLE BIT

55
00:01:57,560 --> 00:01:58,960
EXCITING AND NERVE-WRACKING TO

56
00:01:58,960 --> 00:01:59,800
PRESENT IN FRONT OF YOUR

57
00:01:59,800 --> 00:02:00,600
COLLEAGUES BECAUSE YOU'RE GOING

58
00:02:00,600 --> 00:02:02,640
TO SEE THEM AGAIN AND AGAIN.

59
00:02:02,640 --> 00:02:03,520
AND SO THEY WILL TEND TO

60
00:02:03,520 --> 00:02:04,800
REMEMBER THE MISTAKES YOU MADE,

61
00:02:04,800 --> 00:02:06,600
NOT THE POSITIVE THINGS, SO BEAR

62
00:02:06,600 --> 00:02:10,240
WITH ME AND FORGIVE ME IF I MAKE

63
00:02:10,240 --> 00:02:10,840
ANY MISTAKES.

64
00:02:10,840 --> 00:02:12,920
SO I WANT TO FIRST SAY THAT I'M

65
00:02:12,920 --> 00:02:14,640
VERY EXCITED AND THRILLED TO

66
00:02:14,640 --> 00:02:16,880
BECOME PART OF THE INTRAMURAL

67
00:02:16,880 --> 00:02:17,760
PROGRAM HERE AT NCI.

68
00:02:17,760 --> 00:02:19,680
IT WAS AN EYE-OPENING EXPERIENCE

69
00:02:19,680 --> 00:02:21,400
DURING THE INTERVIEW PROCESS,

70
00:02:21,400 --> 00:02:23,640
AND AFTER COMING HERE, IT'S ONLY

71
00:02:23,640 --> 00:02:25,680
GROWN ON ME MORE THAN IT DID

72
00:02:25,680 --> 00:02:26,040
BEFORE.

73
00:02:26,040 --> 00:02:27,120
SO I WANT TO THANK THE COMMITTEE

74
00:02:27,120 --> 00:02:29,160
AND TOM FOR TRUSTING IN ME AND

75
00:02:29,160 --> 00:02:30,040
RECRUITING ME HERE BEFORE I

76
00:02:30,040 --> 00:02:33,160
START TALKING ABOUT WHAT WE DO.

77
00:02:33,160 --> 00:02:34,960
SO WHAT I THOUGHT I'LL DO IS TO

78
00:02:34,960 --> 00:02:36,600
SUMMARIZE SOME OF THE FINDINGS

79
00:02:36,600 --> 00:02:40,040
FROM MY LAB THAT GIVES A FLAVOR

80
00:02:40,040 --> 00:02:42,800
FOR THE KIND OF RESEARCH WE DO,

81
00:02:42,800 --> 00:02:45,040
AND I WANT TO START HERE SAYING

82
00:02:45,040 --> 00:02:47,240
THAT THE OVERALL -- IN PRINCIPLE

83
00:02:47,240 --> 00:02:48,600
THE OVERALL RESEARCH PROGRAM IN

84
00:02:48,600 --> 00:02:51,080
MY LAB REALLY IS AN INTEGRATION

85
00:02:51,080 --> 00:02:52,320
BETWEEN BASIC AND TRANSLATIONAL

86
00:02:52,320 --> 00:02:54,280
SCIENCE.

87
00:02:54,280 --> 00:02:55,200
THE BASIC SCIENCE SIDE OF IT

88
00:02:55,200 --> 00:02:56,560
REALLY DEALS WITH CELL PRO LAIRT

89
00:02:56,560 --> 00:03:02,160
PROPOLARITYPROTEINS AND THE ROLG

90
00:03:02,160 --> 00:03:03,960
RESISTANCE AND THE TRANSLATION

91
00:03:03,960 --> 00:03:06,480
SIDE DEALS WITH 3D CULTURE AND

92
00:03:06,480 --> 00:03:08,240
ORGANOID MODELS IN THE CONTEXT

93
00:03:08,240 --> 00:03:12,080
OF TRANSLATION RESEARCH.

94
00:03:12,080 --> 00:03:14,240
SO FOR THE SAKE OF STRUCTURE AND

95
00:03:14,240 --> 00:03:15,840
ORGANIZATION, I'M GOING TO

96
00:03:15,840 --> 00:03:17,840
REALLY GROUP MY TALK IN TWO

97
00:03:17,840 --> 00:03:19,720
GROUPS, ON TWO TOPIC.

98
00:03:19,720 --> 00:03:21,520
THE TOPIC ONE DEALS WITH THE

99
00:03:21,520 --> 00:03:22,880
MODELS WE GENERATE AND HOW WE

100
00:03:22,880 --> 00:03:24,640
USE THEM FOR TRANSLATION

101
00:03:24,640 --> 00:03:25,920
RESEARCH, AND THEN THE TOPIC TWO

102
00:03:25,920 --> 00:03:27,960
DEALS WITH THE POLARITY AND

103
00:03:27,960 --> 00:03:28,560
MECHANISMS.

104
00:03:28,560 --> 00:03:30,240
AND FEEL FREE TO STOP ME BETWEEN

105
00:03:30,240 --> 00:03:31,400
THE TWO IF YOU NEED TO, BUT I

106
00:03:31,400 --> 00:03:33,720
WILL CONTINUE ON AS IF YOU DON'T

107
00:03:33,720 --> 00:03:36,160
HAVE ANY QUESTIONS AT THAT

108
00:03:36,160 --> 00:03:36,360
POINT.

109
00:03:36,360 --> 00:03:40,200
SO MY INTEREST IN 3D CULTURE

110
00:03:40,200 --> 00:03:41,840
MODELS STARTED WHEN I WAS A GRAD

111
00:03:41,840 --> 00:03:45,200
STUDENT WHEN I REALIZED I -- ANA

112
00:03:45,200 --> 00:03:48,280
POSTDOC AND I REALIZED

113
00:03:48,280 --> 00:03:49,480
EPITHELIAL CELLS AND THE CANCER

114
00:03:49,480 --> 00:03:50,840
THAT COMES WITH IT IN A PATIENT

115
00:03:50,840 --> 00:03:53,520
EXISTS IN A THREE DIMENSIONAL

116
00:03:53,520 --> 00:03:54,480
ORGANIZATION AND GROWING CELLS

117
00:03:54,480 --> 00:03:57,600
IN A PLASTIC DISH DOES NOT

118
00:03:57,600 --> 00:04:00,280
RE-CREATE THE BIOLOGY THAT A

119
00:04:00,280 --> 00:04:02,000
TYPICAL CELL SEES.

120
00:04:02,000 --> 00:04:03,760
SO WE BEGAN A PROJECT TO TAKE

121
00:04:03,760 --> 00:04:08,720
HUMAN MAMMARY CELL LINE CALLED

122
00:04:08,720 --> 00:04:14,280
MCF-10A.

123
00:04:14,280 --> 00:04:15,360
AND DEVELOPED CONDITIONS WHERE

124
00:04:15,360 --> 00:04:17,120
WE CAN PLATE THEM ON MATRIX AND

125
00:04:17,120 --> 00:04:19,040
THEY ACTUALLY OVER A PERIOD OF

126
00:04:19,040 --> 00:04:22,320
ABOUT TWO WEEKS, THEY GROW AND

127
00:04:22,320 --> 00:04:23,080
UNDERGO MORPHOGENESIS TO

128
00:04:23,080 --> 00:04:23,920
ESTABLISH STRUCTURES THAT HAVE A

129
00:04:23,920 --> 00:04:27,600
LOT OF SIMILARITY WITH THAT OF A

130
00:04:27,600 --> 00:04:29,560
HUMAN BREAST ACINI.

131
00:04:29,560 --> 00:04:31,280
THE TOP PART WHERE IT TALKS

132
00:04:31,280 --> 00:04:38,160
ABOUT HOW AN ACINUS LOOKS LIKE

133
00:04:38,160 --> 00:04:39,640
AND A CROSS-SECTION IS VERY

134
00:04:39,640 --> 00:04:42,080
SIMILAR TO WHAT YOU SEE WHEN YOU

135
00:04:42,080 --> 00:04:43,560
GROW CELLS IN 3D CULTURE.

136
00:04:43,560 --> 00:04:45,960
THE LOGIC WAS TO GENERATE THESE

137
00:04:45,960 --> 00:04:47,720
STRUCTURES AND THEN ACTIVATE

138
00:04:47,720 --> 00:04:50,480
ONCOGENES IN THEM SO WE BEGIN TO

139
00:04:50,480 --> 00:04:51,760
UNDERSTAND HOW THE CELLS WILL

140
00:04:51,760 --> 00:04:54,600
RESPOND IN A THREE DIMENSIONAL

141
00:04:54,600 --> 00:04:56,160
CONTEXT AS OPPOSED TO A REGULAR

142
00:04:56,160 --> 00:04:56,600
MONO LAYER.

143
00:04:56,600 --> 00:04:57,440
THE RESULTS WERE QUITE

144
00:04:57,440 --> 00:04:58,760
INTERESTING AND REALLY

145
00:04:58,760 --> 00:05:01,640
FASCINATING FOR US PAU PAUSE WHE

146
00:05:01,640 --> 00:05:06,640
TAKE THESE CELLS AND ACTIVATE AN

147
00:05:06,640 --> 00:05:09,280
ONCOGENE, ERBB2, ONE OF THE

148
00:05:09,280 --> 00:05:11,200
DOMINANT ONCOGENES IN BREAST

149
00:05:11,200 --> 00:05:12,320
CANCER AND THERE ARE MANY DRUGS

150
00:05:12,320 --> 00:05:14,360
THAT ARE CURRENTLY USED TO TREAT

151
00:05:14,360 --> 00:05:18,360
PATIENTS WITH THIS POSITIVE

152
00:05:18,360 --> 00:05:19,280
DISEASE THAT EXPRESSES THE

153
00:05:19,280 --> 00:05:19,520
ERBB2.

154
00:05:19,520 --> 00:05:22,960
WHAT WE NOTED WAS THAT WHEN YOU

155
00:05:22,960 --> 00:05:24,400
ACTIVATE ERBB2 THESE THREE

156
00:05:24,400 --> 00:05:25,080
DIMENSIONAL STRUCTURES

157
00:05:25,080 --> 00:05:26,480
COMPLETELY LOSE CONTROL AND GROW

158
00:05:26,480 --> 00:05:28,120
INTO AN AMORPHOUS SORT OF SHAPE

159
00:05:28,120 --> 00:05:31,280
VERY SIMILAR TO HOW CANCEROUS

160
00:05:31,280 --> 00:05:32,680
TISSUES WOULD BE GROWING IN

161
00:05:32,680 --> 00:05:33,040
VIVO.

162
00:05:33,040 --> 00:05:34,240
SO THIS PROMPTED US TO REALLY

163
00:05:34,240 --> 00:05:35,880
THINK ABOUT HOW WE CAN USE THESE

164
00:05:35,880 --> 00:05:37,880
THREE DIMENSIONAL CULTURE MODELS

165
00:05:37,880 --> 00:05:38,960
FOR UNDERSTANDING VARIOUS

166
00:05:38,960 --> 00:05:39,720
ASPECTS OF BIOLOGY.

167
00:05:39,720 --> 00:05:41,160
I WOULD ADD THAT THE SAME CELLS,

168
00:05:41,160 --> 00:05:42,720
WHEN YOU PLATE THEM AS A MONO

169
00:05:42,720 --> 00:05:44,480
LAYER AND ACTIVATE ERBB2, YOU

170
00:05:44,480 --> 00:05:46,040
DON'T REALLY SEE MUCH OF AN

171
00:05:46,040 --> 00:05:47,320
EFFECT OTHER THAN A SMALL AMOUNT

172
00:05:47,320 --> 00:05:48,200
OF PROLIFERATIVE ADVANTAGE.

173
00:05:48,200 --> 00:05:50,600
SO THE BIOLOGY REVEALS ITSELF IN

174
00:05:50,600 --> 00:05:53,360
THE CORRECT CONTEXT THAT IS NOT

175
00:05:53,360 --> 00:05:54,120
APPARENT OTHERWISE.

176
00:05:54,120 --> 00:05:55,760
SO WITH THIS IN MIND, WHEN I

177
00:05:55,760 --> 00:05:57,800
STARTED THE LAB, WE BEGAN USING

178
00:05:57,800 --> 00:05:59,360
THESE MODELS FOR VARIOUS

179
00:05:59,360 --> 00:06:00,400
APPLICATIONS BUT OVER THE YEAR,

180
00:06:00,400 --> 00:06:01,760
WE BEGAN TO THINK ABOUT OTHER

181
00:06:01,760 --> 00:06:02,640
CONTEXTS IN WHICH WE CAN USE

182
00:06:02,640 --> 00:06:03,320
THIS.

183
00:06:03,320 --> 00:06:05,040
AND THAT'S WHEN WE BEGAN

184
00:06:05,040 --> 00:06:07,040
THINKING ABOUT DEVELOPING MODELS

185
00:06:07,040 --> 00:06:10,240
FOR PANCREATIC CANCER USING

186
00:06:10,240 --> 00:06:11,280
EMBRYONIC STEM CELLS.

187
00:06:11,280 --> 00:06:12,840
THE REASON BEING THERE ARE NOT

188
00:06:12,840 --> 00:06:14,680
MANY MODELS AVAILABLE TO STUDY

189
00:06:14,680 --> 00:06:15,760
PANCREATIC CANCER AT THAT TIME,

190
00:06:15,760 --> 00:06:17,120
AND THERE WERE A LOT OF INTEREST

191
00:06:17,120 --> 00:06:19,440
IN GENERATING HUMAN EMBRYONIC

192
00:06:19,440 --> 00:06:21,280
STEM CELLS TO INDUCE THEM TO

193
00:06:21,280 --> 00:06:22,800
DIFFERENTIATE TOWARDS THE

194
00:06:22,800 --> 00:06:23,680
ENDOCRINE PANCREAS.

195
00:06:23,680 --> 00:06:29,040
THESE ARE THE CELLS IN THE

196
00:06:29,040 --> 00:06:30,120
PANCREAS THAT SECRETE INSULIN

197
00:06:30,120 --> 00:06:32,400
AND SO FORTH.

198
00:06:32,400 --> 00:06:33,960
THERE'S A REGION IN THE MEDICINE

199
00:06:33,960 --> 00:06:36,120
FIELD VERY INTERESTED IN

200
00:06:36,120 --> 00:06:37,680
TREATING THEM FOR INSULIN

201
00:06:37,680 --> 00:06:39,000
INSUFFICIENCY.

202
00:06:39,000 --> 00:06:42,160
EXOCRINE PANCREAS IS MADE UP OF

203
00:06:42,160 --> 00:06:44,040
DUCTAL AND EPITHELIAL CELLS, THE

204
00:06:44,040 --> 00:06:46,760
CONTEXT WHERE MOST OF THE

205
00:06:46,760 --> 00:06:47,840
PANCREATIC CANCER ORIGINATES

206
00:06:47,840 --> 00:06:51,720
FROM.

207
00:06:51,720 --> 00:06:54,640
SO LING HUANG IS AT THE FORD

208
00:06:54,640 --> 00:06:55,680
INSTITUTE IN DETROIT,

209
00:06:55,680 --> 00:06:57,040
COORDINATEED WITH A STEM CELL

210
00:06:57,040 --> 00:06:58,160
BIOLOGIST AND SPENT A LOT OF

211
00:06:58,160 --> 00:07:00,440
TIME IN TAKING PANCREATIC

212
00:07:00,440 --> 00:07:02,520
PROGENITOR CELLS AND SLOWLY

213
00:07:02,520 --> 00:07:04,960
COERCING THEM.

214
00:07:04,960 --> 00:07:06,160
THE PROCESS WAS HIGHLY

215
00:07:06,160 --> 00:07:07,360
SUCCESSFUL, IT TOOK A LONG TIME,

216
00:07:07,360 --> 00:07:09,040
BUT IT WAS SUCCESSFUL AND THAT

217
00:07:09,040 --> 00:07:10,000
ACTUALLY LED US INTO REALLY

218
00:07:10,000 --> 00:07:12,560
THINKING THAT WE CAN POTENTIALLY

219
00:07:12,560 --> 00:07:15,000
TAKE STEM CELLS AND PUSH THEM

220
00:07:15,000 --> 00:07:16,440
INTO A LINEAGE OF INTEREST.

221
00:07:16,440 --> 00:07:17,640
THE PROCESS OF DOING THIS REALLY

222
00:07:17,640 --> 00:07:18,840
EDUCATED US IN HOW WE CAN USE

223
00:07:18,840 --> 00:07:20,680
THESE CONDITIONS TO GROW PATIENT

224
00:07:20,680 --> 00:07:22,000
TUMORS DIRECTLY FROM THE

225
00:07:22,000 --> 00:07:23,520
PATIENT, AND THAT LED TO THE

226
00:07:23,520 --> 00:07:25,520
DEVELOPMENT OF A TUMOR ORGANOID

227
00:07:25,520 --> 00:07:27,120
CULTURE CONTEXT FOR PANCREATIC

228
00:07:27,120 --> 00:07:28,800
CANCER WHERE WE CAN TAKE

229
00:07:28,800 --> 00:07:30,600
SURGICALLY RESECTED PANCREAS

230
00:07:30,600 --> 00:07:33,040
TUMORS AND THEN GENERATE TUMOR

231
00:07:33,040 --> 00:07:33,720
ORGANOIDS FROM THEM.

232
00:07:33,720 --> 00:07:36,440
WE NOW USE THESE PLATFORMS QUITE

233
00:07:36,440 --> 00:07:36,720
ROBUSTLY.

234
00:07:36,720 --> 00:07:39,960
THE ORGANOID PLATFORM IS USED

235
00:07:39,960 --> 00:07:41,600
MULTIPLE TIMES, FOR THE STEM

236
00:07:41,600 --> 00:07:45,200
CELL WE STILL STICK TO

237
00:07:45,200 --> 00:07:46,160
PANCREATIC CANCER ORGANOIDS.

238
00:07:46,160 --> 00:07:47,840
FOR THE FIRST ONE, OUR GOAL IS

239
00:07:47,840 --> 00:07:48,960
TO INDUCE DIFFERENTIATION OF

240
00:07:48,960 --> 00:07:50,280
CELLS AND FACILITATE

241
00:07:50,280 --> 00:07:52,480
ORGANIZATION THAT RESEMBLES A

242
00:07:52,480 --> 00:07:53,520
NORMAL EPITHELIAL STRUCTURE.

243
00:07:53,520 --> 00:07:55,640
IN THE TUMOR ORGANOID CASE, THE

244
00:07:55,640 --> 00:07:57,880
GOAL IS TO MAINTAIN

245
00:07:57,880 --> 00:07:58,520
DIFFERENTIATION ON ORGANIZATION

246
00:07:58,520 --> 00:08:00,760
SO THE TUMOR ORGANOIDS WE GROW

247
00:08:00,760 --> 00:08:02,440
IN CULTURE RESEMBLE AS MUCH AS

248
00:08:02,440 --> 00:08:03,440
POSSIBLE THE PATIENT TUMOR IT

249
00:08:03,440 --> 00:08:05,160
CAME FROM.

250
00:08:05,160 --> 00:08:07,800
SO TO MAKE THE POINT VERY CLEAR,

251
00:08:07,800 --> 00:08:09,440
SO IN THE TOP PANEL, WHAT YOU

252
00:08:09,440 --> 00:08:10,960
SEE IS THAT THESE ARE EMBRYONIC

253
00:08:10,960 --> 00:08:14,000
STEM CELLS INDUCED TO

254
00:08:14,000 --> 00:08:17,640
DIFFERENTIATE INTO DUCTAL

255
00:08:17,640 --> 00:08:18,840
ACINUS, THEY'RE MUCH SMALLER

256
00:08:18,840 --> 00:08:19,920
THAN THE DUCTAL ORGANOIDS.

257
00:08:19,920 --> 00:08:21,840
THIS IS WHAT YOU SEE IN VIVO,

258
00:08:21,840 --> 00:08:24,520
AND THEY HAVE PROPER APICAL

259
00:08:24,520 --> 00:08:26,920
BASAL POLARIZATION OF BASE MEMO

260
00:08:26,920 --> 00:08:27,680
BRAINGS IN THE BASAL SIDE AND

261
00:08:27,680 --> 00:08:30,120
TIGHT JUNCTIONS IN THE MIDDLE

262
00:08:30,120 --> 00:08:31,120
SURROUNDING THE LUMEN.

263
00:08:31,120 --> 00:08:34,160
THE RIGHT PANEL IS AN ELECTRON

264
00:08:34,160 --> 00:08:35,000
MICROGRAPH SHOWING YOU HOW

265
00:08:35,000 --> 00:08:36,080
THEY'RE ORGANIZED.

266
00:08:36,080 --> 00:08:37,240
TUMORS ARE ALSO THE SAME.

267
00:08:37,240 --> 00:08:38,760
YOU CAN TAKE A PATIENT TUMOR,

268
00:08:38,760 --> 00:08:40,960
GROW THEM IN ORGANOIDS, AS

269
00:08:40,960 --> 00:08:42,240
ORGANOIDS, THEN IF YOU OBSERVE

270
00:08:42,240 --> 00:08:43,480
THE HISTOPATHOLOGY OF THE

271
00:08:43,480 --> 00:08:45,720
ORGANOID THAT WE GROW, THE

272
00:08:45,720 --> 00:08:49,200
ARCHITECTURE OF THE ORGANOID HAS

273
00:08:49,200 --> 00:08:51,160
AN UNCANNY RESEMBLANCE TO WHERE

274
00:08:51,160 --> 00:08:52,360
THE PATIENT TUMOR CAME FROM.

275
00:08:52,360 --> 00:08:55,160
SO THAT GAVE US THE CONFIDENCE

276
00:08:55,160 --> 00:08:56,400
THAT THE CULTURE CONDITIONS WE

277
00:08:56,400 --> 00:08:58,160
HAVE GENERATED ALLOWS THE CELLS

278
00:08:58,160 --> 00:08:59,680
TO EXIST IN A STRUCTURE AND

279
00:08:59,680 --> 00:09:01,280
FUNCTION FORMAT THAT IS SIMILAR

280
00:09:01,280 --> 00:09:03,120
TO WHAT IS EXISTING IN VIVO,

281
00:09:03,120 --> 00:09:04,920
WHICH MEANS THEY COULD BE REALLY

282
00:09:04,920 --> 00:09:06,320
POWERFUL TOOLS TO REALLY

283
00:09:06,320 --> 00:09:08,560
UNDERSTAND CANCER BIOLOGY MORE.

284
00:09:08,560 --> 00:09:11,360
SO WITH THESE MODELS IN MIND, WE

285
00:09:11,360 --> 00:09:12,600
HAVE DONE A SERIES OF

286
00:09:12,600 --> 00:09:14,160
EXPERIMENTS WHERE WE USE THEM

287
00:09:14,160 --> 00:09:16,000
FOR VARIOUS APPLICATIONS, BOTH

288
00:09:16,000 --> 00:09:17,040
THE DISCOVERY AND FOR

289
00:09:17,040 --> 00:09:18,400
TRANSLATION RESEARCH, AND FOR

290
00:09:18,400 --> 00:09:21,640
THE SAKE OF CLARITY, I'M GOING

291
00:09:21,640 --> 00:09:23,160
TO GROUP THE RESULTS IN THREE

292
00:09:23,160 --> 00:09:24,640
CATEGORIES AND PRESENT TO YOU

293
00:09:24,640 --> 00:09:26,240
AND SUMMARIZE FOR THE MOST PART

294
00:09:26,240 --> 00:09:27,840
THE RESULTS WE HAVE OBTAINED TO

295
00:09:27,840 --> 00:09:29,480
MAKE THE POINTS ON HOW WE USE

296
00:09:29,480 --> 00:09:31,240
STEM CELL ORGANOIDS FOR MODELING

297
00:09:31,240 --> 00:09:32,920
CANCER INITIATION, HOW WE USE

298
00:09:32,920 --> 00:09:34,400
TUMOR ORGANOIDS IN THE CONTEXT

299
00:09:34,400 --> 00:09:37,280
OF PRE-CLINICAL AND CLINICAL

300
00:09:37,280 --> 00:09:39,120
STUDIES, AND LASTLY, ONE OF THE

301
00:09:39,120 --> 00:09:40,760
MORE EXCITING AVENUES FOR US IS

302
00:09:40,760 --> 00:09:43,240
HOW WE CAN USE ORGANOIDS TO

303
00:09:43,240 --> 00:09:44,960
IDENTIFY AND DISCOVER NEW CLASS

304
00:09:44,960 --> 00:09:46,560
OF TUMOR TARGETING T-CELL

305
00:09:46,560 --> 00:09:48,200
RECEPTORS.

306
00:09:48,200 --> 00:09:50,520
SO FOR THE FIRST ONE, OUR FOCUS

307
00:09:50,520 --> 00:09:52,040
WAS ON PANCREATIC CANCER BECAUSE

308
00:09:52,040 --> 00:09:53,360
IT IS AN UNMET NEED.

309
00:09:53,360 --> 00:09:56,040
AS YOU KNOW, THAT MOST OF THE

310
00:09:56,040 --> 00:09:56,800
PANCREATIC CANCER PATIENTS ARE

311
00:09:56,800 --> 00:09:57,400
DIAGNOSED LATE.

312
00:09:57,400 --> 00:09:58,680
WE DO NOT KNOW HOW TO DETECT

313
00:09:58,680 --> 00:10:00,880
THEM EARLY SO UNDERSTANDING HOW

314
00:10:00,880 --> 00:10:02,960
CANCER ORIGINATES IN PANCREATIC

315
00:10:02,960 --> 00:10:04,280
CANCER CAN LIKELY PROVIDE NEW

316
00:10:04,280 --> 00:10:07,080
INSIGHTS AN WITH THAT IN MIND,

317
00:10:07,080 --> 00:10:09,040
WE WENT BACK TO OUR STEM CELL

318
00:10:09,040 --> 00:10:10,840
BASED EXOCRINE PANCREAS ORGANOID

319
00:10:10,840 --> 00:10:12,040
MODEL AND BEGAN USING THEM.

320
00:10:12,040 --> 00:10:15,160
I'LL JUST SHOW YOU ONE PIECE OF

321
00:10:15,160 --> 00:10:16,680
DATA TO COMMUNICATE TO YOU THAT

322
00:10:16,680 --> 00:10:17,960
IN IN FACT THESE LINEAGE

323
00:10:17,960 --> 00:10:19,360
COMMITTED ORGANOIDS IN FACT ARE

324
00:10:19,360 --> 00:10:20,320
TRULY LINEAGE COMMITTED.

325
00:10:20,320 --> 00:10:22,600
WE HAVE DONE EXTENSIVE ANALYSIS

326
00:10:22,600 --> 00:10:24,680
BY GENE EXPRESSION COMPARISON

327
00:10:24,680 --> 00:10:26,440
BETWEEN ACINAL AND DUCTAL BUT TO

328
00:10:26,440 --> 00:10:27,520
MAKE THE POINT, I'LL SHOW YOU

329
00:10:27,520 --> 00:10:28,960
SOME OF THE READOUTS THAT WE

330
00:10:28,960 --> 00:10:29,520
HAVE USED AS WELL.

331
00:10:29,520 --> 00:10:32,560
IF YOU LOOK AT MOLECULAR

332
00:10:32,560 --> 00:10:33,280
MARKERS, FOR EXAMPLE, DUCTAL

333
00:10:33,280 --> 00:10:37,040
CELLS ARE KNOWN TO EXPRESS SOCKS

334
00:10:37,040 --> 00:10:44,920
SOX9 AND -- WHICH IS

335
00:10:44,920 --> 00:10:47,200
UNDETECTABLE IN THE ACINAR

336
00:10:47,200 --> 00:10:47,480
LINEAGE.

337
00:10:47,480 --> 00:10:53,360
THE OPPOSITE IS TRUE -- YOU WILL

338
00:10:53,360 --> 00:10:56,240
SEE THE ACINUS ORGANOIDS EXPRESS

339
00:10:56,240 --> 00:10:58,600
THEM QUITE READILY WHEREAS THE

340
00:10:58,600 --> 00:11:00,720
DUCTAL ONES DO NOT.

341
00:11:00,720 --> 00:11:01,800
FUNCTIONALLY THESE ORGANOIDS

342
00:11:01,800 --> 00:11:03,440
ALSO RETAIN THEIR

343
00:11:03,440 --> 00:11:05,040
LINEAGE-SPECIFIC FUNCTION

344
00:11:05,040 --> 00:11:08,280
BECAUSE IF YOU MEASURE CARBONIC

345
00:11:08,280 --> 00:11:10,880
OR LIPASE ACTIVITY, ACINAR CELLS

346
00:11:10,880 --> 00:11:14,760
ARE KNOWN TO SECRETE -- IN OUR

347
00:11:14,760 --> 00:11:16,720
BODY, HIGH IN THE DUCTAL AND LOW

348
00:11:16,720 --> 00:11:20,480
IN ACINAR, THE OPPOSITE IS TRUE

349
00:11:20,480 --> 00:11:23,880
FOR LIGHT-BASED ACTIVITY.

350
00:11:23,880 --> 00:11:24,920
THESE ORGANS WE GENERATE DO HAVE

351
00:11:24,920 --> 00:11:28,360
A LINEAGE COMMITMENT WI BUT FROA

352
00:11:28,360 --> 00:11:29,200
MOLECULAR AND FUNCTIONAL POINT

353
00:11:29,200 --> 00:11:29,560
OF VIEW.

354
00:11:29,560 --> 00:11:30,960
THE QUESTION IS WHAT CAN WE USE

355
00:11:30,960 --> 00:11:31,520
THESE FOR?

356
00:11:31,520 --> 00:11:33,800
NOW THERE ARE TWO TYPES OF

357
00:11:33,800 --> 00:11:35,000
PANCREATIC PRE-MALIGNANT

358
00:11:35,000 --> 00:11:35,200
LESIONS.

359
00:11:35,200 --> 00:11:36,640
ONE OF THEM WHICH IS THE MORE

360
00:11:36,640 --> 00:11:39,200
COMMON ONE IS CALLED THE

361
00:11:39,200 --> 00:11:43,520
PANCREATIC INTRA -- IT'S THOUGHT

362
00:11:43,520 --> 00:11:47,280
TO ORIGINATE FROM DUCTAL OR

363
00:11:47,280 --> 00:11:52,640
ACINAR OR CENTROACINAR CELLS --

364
00:11:52,640 --> 00:11:54,080
THESE CELLS WHEN THEY'RE

365
00:11:54,080 --> 00:11:57,200
TRANSFORMED AND THE DRIVER

366
00:11:57,200 --> 00:12:02,360
ONCOGENE IS MUTANT -- SO WE

367
00:12:02,360 --> 00:12:03,760
DESIGNED A STRATEGY SO WE CAN

368
00:12:03,760 --> 00:12:08,960
USE AN INDUCIBLE -- KRAS, ALLOW

369
00:12:08,960 --> 00:12:10,360
TO DIFFERENTIATE INTO DIFFERENT

370
00:12:10,360 --> 00:12:12,240
LINEAGES AND THEN YOU ACTIVATE

371
00:12:12,240 --> 00:12:14,200
KRAS AND ASK WHAT HAPPENS TO THE

372
00:12:14,200 --> 00:12:14,440
PHENOTYPE.

373
00:12:14,440 --> 00:12:15,840
THE RESULTS ARE PUBLISHED, I

374
00:12:15,840 --> 00:12:19,480
WON'T GO INTO THE DETAILS.

375
00:12:19,480 --> 00:12:22,080
WHAT WE FOUND WAS THAT BOTH

376
00:12:22,080 --> 00:12:24,600
ACINAR AND DUCTAL CELLS DO

377
00:12:24,600 --> 00:12:26,880
RESPOND TO KRAS -- UNDERGOING

378
00:12:26,880 --> 00:12:28,160
DISRUPTION OF THE THREE

379
00:12:28,160 --> 00:12:30,440
DIMENSIONAL STRUCTURE.

380
00:12:30,440 --> 00:12:34,000
INTERESTINGLY, ACINAR CELLS WHEN

381
00:12:34,000 --> 00:12:39,360
YOU ACTIVATE KRAS UNDERGO

382
00:12:39,360 --> 00:12:39,960
METAPLASIA.

383
00:12:39,960 --> 00:12:42,280
IT'S THE WAY THE ACINAR CELLS

384
00:12:42,280 --> 00:12:43,840
WILL NOW BECOME TUMORS.

385
00:12:43,840 --> 00:12:47,680
THEY WILL GO ON TO TRANSFORM TO

386
00:12:47,680 --> 00:12:50,880
BECOME DUCTAL CARCINOMA.

387
00:12:50,880 --> 00:12:51,840
WE OBSERVED THE SAME THING IN

388
00:12:51,840 --> 00:12:52,400
CULTURE AS WELL.

389
00:12:52,400 --> 00:12:53,560
MORE INTERESTINGLY IF YOU TAKE

390
00:12:53,560 --> 00:12:54,840
THESE CELLS AND YOU TRANSPLANT

391
00:12:54,840 --> 00:12:58,920
THEM INTO THE PANCREAS OF MICE,

392
00:12:58,920 --> 00:13:05,960
THE KRAS G12D EXPRESSING ACINAR

393
00:13:05,960 --> 00:13:07,280
CELLS -- WHEREAS THE SAME

394
00:13:07,280 --> 00:13:09,560
ONCOGENE EXPRESSING ACINAR CELLS

395
00:13:09,560 --> 00:13:11,360
WERE ONLY 11% EFFICIENT.

396
00:13:11,360 --> 00:13:13,000
THIS DICHOTOMY IN THE WAY IN

397
00:13:13,000 --> 00:13:15,400
WHICH KRAS WORKS IN ACINAR

398
00:13:15,400 --> 00:13:16,880
VERSUS DUCTAL IS EXACTLY WHAT

399
00:13:16,880 --> 00:13:18,280
IT'S BEEN REPORTED IN MULTIPLE

400
00:13:18,280 --> 00:13:19,640
STUDIES IN MOUSE MODELS, AND

401
00:13:19,640 --> 00:13:24,480
THOSE ARE STUDIES THAT I POINT

402
00:13:24,480 --> 00:13:25,800
HERE -- I THINK THIS REALLY

403
00:13:25,800 --> 00:13:27,520
CONFIRMS TO US THESE TYPE OF

404
00:13:27,520 --> 00:13:30,640
LINEAGE COMMITTED ORGANOIDS ARE

405
00:13:30,640 --> 00:13:31,720
POWERFUL TOOL TO REALLY

406
00:13:31,720 --> 00:13:34,200
UNDERSTAND HOW THE CANCER

407
00:13:34,200 --> 00:13:36,840
INITIATES IN THIS MODEL SYSTEM.

408
00:13:36,840 --> 00:13:38,160
THE ADVANTAGE BETWEEN MOUSE AND

409
00:13:38,160 --> 00:13:39,920
HERE IS WE KNOW THERE IS

410
00:13:39,920 --> 00:13:40,560
SIGNIFICANT DIFFERENCE BETWEEN

411
00:13:40,560 --> 00:13:41,920
MOUSE AND HUMAN CELL LINES SO

412
00:13:41,920 --> 00:13:44,200
HERE WE HAVE A HUMAN CELL LINE

413
00:13:44,200 --> 00:13:45,920
BASED MODEL THAT WE CAN USE FOR

414
00:13:45,920 --> 00:13:47,200
UNDERSTANDING EARLY STAGES.

415
00:13:47,200 --> 00:13:50,440
SO LING HUANG, WHO IS NOW IN

416
00:13:50,440 --> 00:13:52,600
HENRY FORD HEALTH, WILL NOW BE

417
00:13:52,600 --> 00:13:55,120
PURSUING MANY OF THESE STUDIES

418
00:13:55,120 --> 00:13:56,520
IN HIS OWN GROUP IN DETROIT.

419
00:13:56,520 --> 00:13:59,080
SO THE SECOND FORM OF LESION

420
00:13:59,080 --> 00:14:00,440
THAT IS LESS FREQUENT BUT IT IS

421
00:14:00,440 --> 00:14:01,680
OBSERVED IN PATIENTS IN A

422
00:14:01,680 --> 00:14:04,120
SIGNIFICANT NUMBER ARE CALLED

423
00:14:04,120 --> 00:14:05,360
INTRAPAPILLARY MUCINOUS

424
00:14:05,360 --> 00:14:06,200
NEOPLASMS.

425
00:14:06,200 --> 00:14:07,680
UNLIKE IPM AND THOSE ARE

426
00:14:07,680 --> 00:14:09,040
MICROSCOPIC IN THE PANCREAS,

427
00:14:09,040 --> 00:14:10,760
THESE ARE LARGE CYSTIC

428
00:14:10,760 --> 00:14:11,400
DILATIONS.

429
00:14:11,400 --> 00:14:13,800
THAT MEANS THAT THESE ARE

430
00:14:13,800 --> 00:14:15,680
DETECTABLE BY MOLECULAR IMAGING

431
00:14:15,680 --> 00:14:17,200
IN A PATIENT, SO THERE ARE MANY

432
00:14:17,200 --> 00:14:19,200
PATIENTS, HUNDREDS AND THOUSANDS

433
00:14:19,200 --> 00:14:20,800
OF PATIENTS TODAY WALKING AROUND

434
00:14:20,800 --> 00:14:24,240
WITH AN IPMN LESION WHO ARE JUST

435
00:14:24,240 --> 00:14:25,520
MONITORED CLINICALLY TO SEE IF

436
00:14:25,520 --> 00:14:28,400
AND WHEN THEY WILL PROGRESS.

437
00:14:28,400 --> 00:14:30,680
SO IF YOU CAN UNDERSTAND HOW

438
00:14:30,680 --> 00:14:31,640
THEY'RE REGULATED YOU CAN

439
00:14:31,640 --> 00:14:33,040
ACTUALLY HAVE A THERAPEUTIC

440
00:14:33,040 --> 00:14:35,840
IMPACT TO PREVENT PROGRESSION OF

441
00:14:35,840 --> 00:14:38,000
PATIENTS WITH IPMN LESIONS.

442
00:14:38,000 --> 00:14:40,120
THE CELL OF ORIGIN FOR -- IS

443
00:14:40,120 --> 00:14:42,640
THOUGHT TO BE DUCTAL ALTHOUGH

444
00:14:42,640 --> 00:14:44,560
THERE'S NO CLEAR UNTIL OUR STUDY

445
00:14:44,560 --> 00:14:46,240
WAS PUBLISHED.

446
00:14:46,240 --> 00:14:51,240
THERE IS A MUTATION CALLED GNAS,

447
00:14:51,240 --> 00:14:54,400
ALWAYS SEEN IN EARLY IPMN

448
00:14:54,400 --> 00:14:55,840
LESIONS.

449
00:14:55,840 --> 00:14:59,360
SO WE DECIDED TO TAKE THE GNAS

450
00:14:59,360 --> 00:15:01,120
MUTANT AND DO THE SAME LOGIC AS

451
00:15:01,120 --> 00:15:06,480
WE DID IN KRAS AND ASK WHAT

452
00:15:06,480 --> 00:15:06,680
HAPPENS.

453
00:15:06,680 --> 00:15:12,520
SO I W A BOA POSTDOC IN THE LABD

454
00:15:12,520 --> 00:15:15,200
THESE EXPERIMENTS USING THE MU

455
00:15:15,200 --> 00:15:17,640
TAN GNAS IN DUCTAL OR ACINAR

456
00:15:17,640 --> 00:15:17,960
CELLS.

457
00:15:17,960 --> 00:15:18,960
WHAT YOU NOTICE ON THE LEFT IS

458
00:15:18,960 --> 00:15:22,120
WHEN YOU ACTIVATE GNAS IN THE

459
00:15:22,120 --> 00:15:25,520
DUCTAL CELLS YOU GET THESE LARGE

460
00:15:25,520 --> 00:15:28,000
CYSTIC DILATIONS IN GROWTH.

461
00:15:28,000 --> 00:15:29,640
THERE'S INCREASE IN CELL NUMBER

462
00:15:29,640 --> 00:15:32,840
AND STRUCTURE VO VOLUME.

463
00:15:32,840 --> 00:15:34,800
WHEREAS ACINAR CELLS ON THE

464
00:15:34,800 --> 00:15:36,880
OTHER HAND SHOW A MODEST

465
00:15:36,880 --> 00:15:37,440
PHENOTYPE.

466
00:15:37,440 --> 00:15:38,320
THE SIZE DIFFERENCE IS RATHER

467
00:15:38,320 --> 00:15:38,960
SMALL.

468
00:15:38,960 --> 00:15:40,920
YOU CAN QUANTITATIVELY OBSERVE

469
00:15:40,920 --> 00:15:42,680
THERE'S AN EIGHT FOLD DIFFERENCE

470
00:15:42,680 --> 00:15:44,880
IN CELL SIZE IN RESPONSE TO GNAS

471
00:15:44,880 --> 00:15:46,720
IN DUCTAL WHERE IT'S LESS THAN

472
00:15:46,720 --> 00:15:49,160
THREE FOLD IN ACINAR ORGANOIDS.

473
00:15:49,160 --> 00:15:51,320
SO THIS DIFFERENTIAL RESPONSE TO

474
00:15:51,320 --> 00:15:53,000
IT WAS ACTUALLY QUITE

475
00:15:53,000 --> 00:15:53,320
INTERESTING.

476
00:15:53,320 --> 00:15:54,400
EVEN MORE INTERESTING WAS THAT

477
00:15:54,400 --> 00:15:56,920
IF YOU LOOK AT EXPRESSION OF

478
00:15:56,920 --> 00:15:58,280
MOLECULAR MARKER THAT IS

479
00:15:58,280 --> 00:16:01,720
ROUTINELY USED TO DIAGNOSE IPMN

480
00:16:01,720 --> 00:16:02,480
IN THE CLINIC.

481
00:16:02,480 --> 00:16:08,400
SO EXPRESSION OF MUC2, THE DUCTS

482
00:16:08,400 --> 00:16:10,200
THAT ARE DILATED, IS A MARKER

483
00:16:10,200 --> 00:16:12,120
FOR IPMN LESIONS.

484
00:16:12,120 --> 00:16:14,000
WHEN WE ACTIVATED GNAS IN THE

485
00:16:14,000 --> 00:16:15,640
DUCTAL ORGANOIDS, WE CAN SEE

486
00:16:15,640 --> 00:16:19,120
EXPRESSION OF MUC2 WHEREAS THAT

487
00:16:19,120 --> 00:16:20,440
IS NOT OBSERVED WHEN YOU

488
00:16:20,440 --> 00:16:23,440
ACTIVATE THEM IN ACINAR

489
00:16:23,440 --> 00:16:25,040
ORGANOIDS.

490
00:16:25,040 --> 00:16:26,040
WE CLEARLY DEMONSTRATE IN THIS

491
00:16:26,040 --> 00:16:27,920
THAT ACINAR CELLS CANNOT

492
00:16:27,920 --> 00:16:30,000
POSSIBLY GO DOWN THE PATH OF

493
00:16:30,000 --> 00:16:31,400
IPMN UNDER THESE CONDITIONS.

494
00:16:31,400 --> 00:16:36,800
IF YOU FORCE THEM, ACINAR DUCTAL

495
00:16:36,800 --> 00:16:40,600
METAPLASIA AND THEN EXPRESS MUCM

496
00:16:40,600 --> 00:16:41,440
THIS WHOLE PROCESS.

497
00:16:41,440 --> 00:16:43,360
SO WE REALLY THOUGHT WITH ABOUT

498
00:16:43,360 --> 00:16:44,760
OKAY HOW DO WE REALLY UNDERSTAND

499
00:16:44,760 --> 00:16:46,280
THIS BIOLOGY AND WHAT ARE THE

500
00:16:46,280 --> 00:16:47,520
MOLECULAR MECHANISMS BY WHICH

501
00:16:47,520 --> 00:16:48,680
THIS IS BEING REGULATED.

502
00:16:48,680 --> 00:16:50,720
SO IT'S A G PROTEIN COUPLED

503
00:16:50,720 --> 00:16:51,320
RECEPTOR.

504
00:16:51,320 --> 00:16:54,200
ALMOST ALL KNOWN FUNCTIONS OF

505
00:16:54,200 --> 00:16:55,600
GNAS IS THOUGHT TO GO THROUGH

506
00:16:55,600 --> 00:16:57,640
ACTIVATION OF PKA, SO WE THOUGHT

507
00:16:57,640 --> 00:16:59,920
OKAY IF WE INHIBIT PKA WE SHOULD

508
00:16:59,920 --> 00:17:02,080
BE ABLE TO INHIBIT ALL THE

509
00:17:02,080 --> 00:17:07,400
BIOLOGY DOWNSTREAM OF GNAS.

510
00:17:07,400 --> 00:17:10,560
SO WE USED A FEW DIFFERENT

511
00:17:10,560 --> 00:17:12,240
INHIBITORS, I'M SHOWING DATA FOR

512
00:17:12,240 --> 00:17:14,480
ONE OF THEM.

513
00:17:14,480 --> 00:17:16,560
YOU LOOK FOR GNAS-INDUCED

514
00:17:16,560 --> 00:17:20,320
SIGNALING BY READING OUT EITHER

515
00:17:20,320 --> 00:17:22,320
PAN PKA SUBSTRATE ANTIBODY ON

516
00:17:22,320 --> 00:17:25,080
THE TOP OR A SPECIFIC SUBSTRATE

517
00:17:25,080 --> 00:17:27,560
OR PVASP ON THE BOTTOM, YOU CAN

518
00:17:27,560 --> 00:17:28,680
COMPLETELY SHUT DOWN PK

519
00:17:28,680 --> 00:17:30,280
SIGNALING IN THESE CELLS.

520
00:17:30,280 --> 00:17:31,520
SO WHAT HAPPENS IF YOU ADD THE

521
00:17:31,520 --> 00:17:32,560
INHIBITOR AND THEN ACTIVATE IN

522
00:17:32,560 --> 00:17:34,560
THESE DUCTAL LESIONS?

523
00:17:34,560 --> 00:17:43,520
SURPRISINGLY IT HAD NO EFFECT.

524
00:17:43,520 --> 00:17:44,640
SO I REALLY WANTED TO TEST

525
00:17:44,640 --> 00:17:46,600
WHETHER THIS IS ACTUALLY -- THIS

526
00:17:46,600 --> 00:17:48,920
IS A PROLIFERATION-DRIVEN OR

527
00:17:48,920 --> 00:17:52,320
IT'S CYSTIC DILATION BECAUSE

528
00:17:52,320 --> 00:17:52,800
OF -- ACTIVITY.

529
00:17:52,800 --> 00:17:55,720
SO WE DID SOME EDU LABELING AND

530
00:17:55,720 --> 00:17:56,760
THEN ASKED IS THE LABELING

531
00:17:56,760 --> 00:17:59,320
HAPPENING IN RESPONSE TO GNAS

532
00:17:59,320 --> 00:18:00,280
ACTIVATION IN THE ABSENCE AND

533
00:18:00,280 --> 00:18:01,800
PRESENCE OF THE INHIBITOR, AND

534
00:18:01,800 --> 00:18:05,840
IN BOTH CASES -- WAS ABLE TO SEE

535
00:18:05,840 --> 00:18:09,000
INITIATION OF DNA SYNTHESIS IN

536
00:18:09,000 --> 00:18:11,400
BOTH -- IN THE DUCTAL ORGANOIDS.

537
00:18:11,400 --> 00:18:12,640
AND WHEN SHE QUANTITY TAITS IT,

538
00:18:12,640 --> 00:18:19,240
QUANTITATES IT, ONTHE LEFT, IN T

539
00:18:19,240 --> 00:18:22,000
HAD NO EFFECT IN THE FUNCTION OF

540
00:18:22,000 --> 00:18:22,640
DNA SYNTHESIS.

541
00:18:22,640 --> 00:18:24,040
SO THIS GAVE US AN IDEA THAT

542
00:18:24,040 --> 00:18:26,120
THERE IS DEFINITELY A

543
00:18:26,120 --> 00:18:27,480
PK-INDEPENDENT ARM OF REGULATING

544
00:18:27,480 --> 00:18:32,040
CELL CYCLE BY GNAS ACTIVATION.

545
00:18:32,040 --> 00:18:35,320
BY CONTRAST, THE ACINAR

546
00:18:35,320 --> 00:18:37,200
ORGANOIDS HAS A WEAK PHENOTYPE,

547
00:18:37,200 --> 00:18:42,560
AND IF YOU'VE SEEN THE BOTTOM IN

548
00:18:42,560 --> 00:18:45,040
THE PRESENCE OF DOX, THAT SMALL

549
00:18:45,040 --> 00:18:46,200
INCREASE IS COMPLETELY SHUT DOWN

550
00:18:46,200 --> 00:18:47,840
WHEN YOU ADD THE PK INHIBITOR.

551
00:18:47,840 --> 00:18:50,480
SO THIS MEANS GNAS IS

552
00:18:50,480 --> 00:18:52,000
EXCLUSIVELY USING DIFFERENT

553
00:18:52,000 --> 00:18:54,200
SIGNALING PATHWAYS AND DIFFERENT

554
00:18:54,200 --> 00:18:57,160
LINEAGES IN HOW IT INITIATES

555
00:18:57,160 --> 00:18:58,560
PROLIFERATION.

556
00:18:58,560 --> 00:19:01,520
THEN WE SAID WHAT HAPPENS TO MUC

557
00:19:01,520 --> 00:19:03,520
MUC2, THIS WAS CONFUSING AND

558
00:19:03,520 --> 00:19:04,360
INTERESTING AT THE SAME TIME.

559
00:19:04,360 --> 00:19:06,120
DUCTAL ORGANOIDS, WAS SET DOWN

560
00:19:06,120 --> 00:19:07,200
BY PKA SIGNALING.

561
00:19:07,200 --> 00:19:08,800
SO THIS IS REALLY PLACED IN A

562
00:19:08,800 --> 00:19:10,520
VERY INTERESTING POSITION

563
00:19:10,520 --> 00:19:12,040
BECAUSE WHAT IT SUGGESTS IS THAT

564
00:19:12,040 --> 00:19:13,160
DEPENDING ON THE LINEAGE AND

565
00:19:13,160 --> 00:19:14,440
DEPENDING ON CONTEXT AND

566
00:19:14,440 --> 00:19:17,000
DEPENDING ON THE READOUT, YOU

567
00:19:17,000 --> 00:19:19,240
MAY -- GNAS'S DIFFERENT

568
00:19:19,240 --> 00:19:21,800
SIGNALING MECHANISMS.

569
00:19:21,800 --> 00:19:22,480
PROLIFERATION IN THE DUCTAL

570
00:19:22,480 --> 00:19:24,080
CELLS IS REALLY DRIVING

571
00:19:24,080 --> 00:19:26,240
PROLIFERATION IN A

572
00:19:26,240 --> 00:19:27,520
PK-INDEPENDENT MANNER WHEREAS

573
00:19:27,520 --> 00:19:29,000
EVERYTHING ELSE WE HAVE READ SO

574
00:19:29,000 --> 00:19:32,400
FAR IS A PK-INDEPENDENT PROCESS.

575
00:19:32,400 --> 00:19:34,160
SO WE NEED TO ASK A SERIES OF

576
00:19:34,160 --> 00:19:37,040
QUESTIONS LIKE HOW DOES GNAS

577
00:19:37,040 --> 00:19:38,320
INDUCE PROLIFERATION IN DUCTAL

578
00:19:38,320 --> 00:19:41,160
CELLS IN PK-INDEPENDENT MANNER.

579
00:19:41,160 --> 00:19:43,120
THE REASON THE ANSWER WOULD BE

580
00:19:43,120 --> 00:19:44,640
QUITE POWERFUL BECAUSE IF YOU

581
00:19:44,640 --> 00:19:47,480
WANT TO INHIBIT -- HAVE GNAS

582
00:19:47,480 --> 00:19:50,560
INDUCED IPMN IN PATIENTS YOU

583
00:19:50,560 --> 00:19:52,080
WANT AN ARM THAT IS GENERALLY

584
00:19:52,080 --> 00:19:53,800
NOT INHIBITING PK TOTAL.

585
00:19:53,800 --> 00:19:54,960
SO IF YOU WANT TO FIND A

586
00:19:54,960 --> 00:19:57,000
SPECIFIC PATHWAY USED TO BIND

587
00:19:57,000 --> 00:19:58,880
GNAS IN DUCTAL CELLS TO INDUCE

588
00:19:58,880 --> 00:20:00,640
PROLIFERATION, THAT COULD HAVE

589
00:20:00,640 --> 00:20:01,520
SIGNIFICANT THERAPEUTIC

590
00:20:01,520 --> 00:20:02,000
POTENTIAL.

591
00:20:02,000 --> 00:20:03,680
AND SHE'S ALSO INTERESTED IN

592
00:20:03,680 --> 00:20:05,240
FINDING OUT WHAT IT TAKES FOR

593
00:20:05,240 --> 00:20:07,720
THESE GNS LESIONS TO BECOME

594
00:20:07,720 --> 00:20:09,400
FULLY MALIGNANT AND SO FORTH.

595
00:20:09,400 --> 00:20:10,520
SHE'S ALSO INTERESTED IN USING

596
00:20:10,520 --> 00:20:12,760
THIS PLATFORM TO IDENTIFY NEW

597
00:20:12,760 --> 00:20:13,680
BIOMARKERS.

598
00:20:13,680 --> 00:20:17,720
IN HER OWN LAB.

599
00:20:17,720 --> 00:20:19,680
SO I HOPE I CONVINCED YOU THAT

600
00:20:19,680 --> 00:20:20,800
THE STEM CELL BASED ORGANOIDS

601
00:20:20,800 --> 00:20:22,080
ARE VERY POWERFUL TOOLS TO

602
00:20:22,080 --> 00:20:23,120
REALLY THINK ABOUT MODELING

603
00:20:23,120 --> 00:20:24,520
DISEASE IN GENERAL, BUT MODELING

604
00:20:24,520 --> 00:20:27,120
CANCER IN OUR CASE.

605
00:20:27,120 --> 00:20:28,400
SO IN THE NEXT FEW MINUTES, I'LL

606
00:20:28,400 --> 00:20:29,680
TALK ABOUT THE WAY IN WHICH WE

607
00:20:29,680 --> 00:20:33,400
USE TUMOR ORGANOIDS FOR VARIOUS

608
00:20:33,400 --> 00:20:36,080
DISCOVERY AND TRANSLATION

609
00:20:36,080 --> 00:20:36,440
APPLICATIONS.

610
00:20:36,440 --> 00:20:37,720
SO WE DID A SERIES OF

611
00:20:37,720 --> 00:20:40,120
EXPERIMENTS THAT I'LL SUMMARIZE

612
00:20:40,120 --> 00:20:42,240
IN THIS BECAUSE I

613
00:20:42,240 --> 00:20:43,680
WANT TO FOCUS IN THE SECOND PART

614
00:20:43,680 --> 00:20:45,520
OF THE RESULTS THAT WE GENERATED

615
00:20:45,520 --> 00:20:47,920
TO UNDERSTAND WHETHER ORGANOIDS

616
00:20:47,920 --> 00:20:53,360
ACTUALLY ARE ABLE TO HAVE

617
00:20:53,360 --> 00:20:55,160
BIOLOGY SIMILAR TO THAT IN VIVO.

618
00:20:55,160 --> 00:20:57,720
WE HAD A SERIES OF PDX TUMORS

619
00:20:57,720 --> 00:21:00,560
AND ORGANOIDS, AND DID A SERIES

620
00:21:00,560 --> 00:21:03,200
OF ANALYSIS INCLUDING GENE

621
00:21:03,200 --> 00:21:05,600
EXPRESSION AND MUTATION PROFILE

622
00:21:05,600 --> 00:21:06,600
ANALYSIS, BUT IN ADDITION TO

623
00:21:06,600 --> 00:21:08,240
THAT, WE DID ALSO A SERIES OF

624
00:21:08,240 --> 00:21:10,000
CELL BIOLOGICAL STUDIES, WHERE

625
00:21:10,000 --> 00:21:13,040
WE COMPARED DRUG SENSITIVITIES,

626
00:21:13,040 --> 00:21:18,080
WE COMPARED THE SECRETORY SYSTEM

627
00:21:18,080 --> 00:21:20,280
IN VIVO COMPARED TO CULTURE AND

628
00:21:20,280 --> 00:21:22,360
ALSO GLYCOMICS, HOW THE SURFACE

629
00:21:22,360 --> 00:21:23,480
PROTEIN ARE CONSERVED BETWEEN

630
00:21:23,480 --> 00:21:26,160
THE ORGANOIDS AND THE PDX

631
00:21:26,160 --> 00:21:26,400
MODELS.

632
00:21:26,400 --> 00:21:27,600
THE RESULTS WERE ACTUALLY QUITE

633
00:21:27,600 --> 00:21:28,160
INFORMATIVE.

634
00:21:28,160 --> 00:21:29,920
WE FOUND THAT THE ORGANOIDS

635
00:21:29,920 --> 00:21:31,440
PRETICKET IN VIVO DRUG RESPONSE

636
00:21:31,440 --> 00:21:35,240
IN ABOUT 80 TO 90% ACCURACY IN

637
00:21:35,240 --> 00:21:36,000
CULTURE.

638
00:21:36,000 --> 00:21:38,520
AND WE ALSO REALIZED THAT

639
00:21:38,520 --> 00:21:39,440
MEASURING DRUG SENSITIVITY IN

640
00:21:39,440 --> 00:21:41,040
ORGANOIDS IS VERY TRICKY.

641
00:21:41,040 --> 00:21:46,120
IT'S NOT EASY TO USE -- EVERY

642
00:21:46,120 --> 00:21:47,560
ORGANOID HAS ITS OWN DOUBLING

643
00:21:47,560 --> 00:21:48,880
TIME AND THERE ARE SLOW GROWING

644
00:21:48,880 --> 00:21:50,040
AND FAST GROWING ONES SO IN

645
00:21:50,040 --> 00:21:51,160
ORDER TO REALLY ACCOUNT FOR

646
00:21:51,160 --> 00:21:52,360
THESE VARIATIONS, WE HAVE

647
00:21:52,360 --> 00:21:53,680
DEVELOPED A NORMALIZED AREA

648
00:21:53,680 --> 00:21:54,880
UNDER THE CURVE METHOD THAT

649
00:21:54,880 --> 00:21:56,800
ALLOWS US TO REALLY OBJECTIVELY

650
00:21:56,800 --> 00:21:58,360
COMPARE DRUG RESPONSE BETWEEN

651
00:21:58,360 --> 00:21:59,040
ORGANOID LINES.

652
00:21:59,040 --> 00:22:00,520
AND WE KNOW NOW THAT THE

653
00:22:00,520 --> 00:22:01,840
VESICLES THAT ARE SECRETED BY

654
00:22:01,840 --> 00:22:03,880
THE ORGANOIDS IN CULTURE CAN BE

655
00:22:03,880 --> 00:22:06,400
SEEN IN THE PATIENT'S BLOOD,

656
00:22:06,400 --> 00:22:07,920
WHICH MEANS THIS COULD BE A

657
00:22:07,920 --> 00:22:14,680
POWERFUL TOOL TO FIND SECRETE --

658
00:22:14,680 --> 00:22:15,840
I THINK THIS CAN BE HELPFUL

659
00:22:15,840 --> 00:22:16,880
THERE.

660
00:22:16,880 --> 00:22:18,880
AND AS I MENTIONED, THE SURFACE

661
00:22:18,880 --> 00:22:20,880
GLYCAN PROFILE ARE ABOUT 80%

662
00:22:20,880 --> 00:22:21,720
SIMILAR, WHICH GIVES US

663
00:22:21,720 --> 00:22:23,960
CONFIDENCE THAT IN THE 3D

664
00:22:23,960 --> 00:22:24,800
CULTURE, THEY'RE ABLE TO

665
00:22:24,800 --> 00:22:27,320
MAINTAIN SOME ASPECT OF THE

666
00:22:27,320 --> 00:22:28,840
SURFACEOME SIMILAR TO THAT IN

667
00:22:28,840 --> 00:22:29,080
VIVO.

668
00:22:29,080 --> 00:22:30,200
SO ARMED WITH THAT INFORMATION,

669
00:22:30,200 --> 00:22:31,520
WE WANTED TO ASK, IS THIS REALLY

670
00:22:31,520 --> 00:22:33,560
GOING TO BE USEFUL IN THE

671
00:22:33,560 --> 00:22:34,560
CLINICAL SETTING?

672
00:22:34,560 --> 00:22:39,480
SO FOR THIS WE NICHE WRA INITIAA

673
00:22:39,480 --> 00:22:40,800
CLINICAL TRIAL CALLED HOPE

674
00:22:40,800 --> 00:22:42,080
REASONS, WE HOPED IT WORKED FOR

675
00:22:42,080 --> 00:22:43,640
THE SAKE OF PATIENTS, AND IT WAS

676
00:22:43,640 --> 00:22:46,480
LED BY JOE GROSSMAN AND MANNY

677
00:22:46,480 --> 00:22:48,920
HIDALGO IN THE CLINIC AND BY

678
00:22:48,920 --> 00:22:51,320
LING HUANG AND -- IN THE LAB,

679
00:22:51,320 --> 00:22:53,080
AND THE GOAL WAS TO GENERATE

680
00:22:53,080 --> 00:22:53,920
ORGANOID MODELS FROM PATIENTS

681
00:22:53,920 --> 00:22:56,520
AND THEN TREAT THE ORGANOID

682
00:22:56,520 --> 00:22:57,920
MODELS WITH THE SAME DRUG

683
00:22:57,920 --> 00:22:59,320
THEY'RE GOING TO RECEIVE AFTER

684
00:22:59,320 --> 00:23:00,400
THE DRUG AND THEN COMPARE THE

685
00:23:00,400 --> 00:23:01,040
RESULTS.

686
00:23:01,040 --> 00:23:02,520
THE END POINTS WERE ACTUALLY

687
00:23:02,520 --> 00:23:04,680
VERY SIMPLE.

688
00:23:04,680 --> 00:23:05,960
WE FIRST WANTED TO KNOW IF THIS

689
00:23:05,960 --> 00:23:07,800
IS FEASIBLE AND SECOND WE WANTED

690
00:23:07,800 --> 00:23:09,480
TO SEE IF THERE WAS A

691
00:23:09,480 --> 00:23:10,720
CORRELATION BETWEEN ORGANOID

692
00:23:10,720 --> 00:23:12,600
RESPONSE IN THE PDO AND PATIENT

693
00:23:12,600 --> 00:23:16,360
RESPONSE IN THE CLINIC.

694
00:23:16,360 --> 00:23:18,720
THE EQUALLY REPRESENTED FROM

695
00:23:18,720 --> 00:23:21,920
MALE AND FEMALE AND ALSO EQUALLY

696
00:23:21,920 --> 00:23:22,840
REPRESENTED FROM ALL STAGES OF

697
00:23:22,840 --> 00:23:26,680
THE DISEASE.

698
00:23:26,680 --> 00:23:29,480
THERE ARE TWO SUBTYPES OF

699
00:23:29,480 --> 00:23:31,000
PANCREATIC CANCER, BASAL AND

700
00:23:31,000 --> 00:23:32,960
CLASSICAL AND WE HAVE ORGANOID

701
00:23:32,960 --> 00:23:35,520
MODELS THAT ARE EITHER CLASSICAL

702
00:23:35,520 --> 00:23:36,960
OR BASAL THAT MATCHES WITH THE

703
00:23:36,960 --> 00:23:39,360
TUMOR AS WELL AND THE CLASSIC

704
00:23:39,360 --> 00:23:41,160
TUMOR RESULTS IN A CLASSIC

705
00:23:41,160 --> 00:23:43,040
ORGANOID AND BASAL PATIENT TUMOR

706
00:23:43,040 --> 00:23:45,000
RESULTS IN A BASAL PART OF OR

707
00:23:45,000 --> 00:23:45,480
THE NOID.

708
00:23:45,480 --> 00:23:46,880
SO IN ORDER TO REALLY UNDERSTAND

709
00:23:46,880 --> 00:23:50,000
HOW THIS IS USEFUL, WE TOOK A

710
00:23:50,000 --> 00:23:51,320
DEEP DIVE APPROACH ON 14

711
00:23:51,320 --> 00:23:51,920
PATIENTS.

712
00:23:51,920 --> 00:23:54,160
NOW THESE 14 PATIENTS RECEIVED

713
00:23:54,160 --> 00:23:55,560
42 DIFFERENT DRUGS, BECAUSE MOST

714
00:23:55,560 --> 00:23:58,360
PATIENTS ARE TREATED WITH

715
00:23:58,360 --> 00:24:00,480
COMBINATIONS AT THIS STAGE, AS

716
00:24:00,480 --> 00:24:01,480
THE COMBINATION LISTED HERE SO

717
00:24:01,480 --> 00:24:04,360
WE DECIDED TO TAKE THESE

718
00:24:04,360 --> 00:24:06,400
ORGANOID MODELS AND GENERATE

719
00:24:06,400 --> 00:24:07,680
42 DIFFERENT AUC VALUES.

720
00:24:07,680 --> 00:24:08,920
THEN WE WENT BACK TO THE CLINIC

721
00:24:08,920 --> 00:24:10,600
AND ASKED WHAT IS THE CLINICAL

722
00:24:10,600 --> 00:24:13,360
RESPONSE OF THIS PATIENTS AS

723
00:24:13,360 --> 00:24:15,000
PERVERSE US TO CRITERIA.

724
00:24:15,000 --> 00:24:17,080
USUALLY A QI NATION OF IMAGING,

725
00:24:17,080 --> 00:24:17,800
BIOMARKERS AND OTHER

726
00:24:17,800 --> 00:24:18,440
CHARACTERISTIC THAT IS USED IN

727
00:24:18,440 --> 00:24:19,240
THE CLINIC.

728
00:24:19,240 --> 00:24:20,800
AND SO WHEN YOU COMPARE THE

729
00:24:20,800 --> 00:24:24,480
PATIENT RESPONSE TO THE AUC

730
00:24:24,480 --> 00:24:25,440
VALUES OF THE DRUG FOR THE

731
00:24:25,440 --> 00:24:26,840
SPECIFIC ORGANOIDS, THERE WAS A

732
00:24:26,840 --> 00:24:28,000
VERY INTERESTING SEGREGATION.

733
00:24:28,000 --> 00:24:30,280
SO THE PATIENTS WHO ARE LABELED

734
00:24:30,280 --> 00:24:33,360
IN BLUE ARE THOSE PATIENTS WHO

735
00:24:33,360 --> 00:24:34,480
HAD YOU HAD EITHER A STABLE

736
00:24:34,480 --> 00:24:35,920
DISEASE OR PARTIAL RESPONSE.

737
00:24:35,920 --> 00:24:38,400
WHEREAS THE PATIENTS IN RED ARE

738
00:24:38,400 --> 00:24:39,520
ALL PATIENTS WHO PROGRESSED ON

739
00:24:39,520 --> 00:24:40,160
THE TREATMENT.

740
00:24:40,160 --> 00:24:42,240
IF YOU LOOK AT ALL THE AUC

741
00:24:42,240 --> 00:24:43,960
VALUES, WHAT WE REALIZED WAS

742
00:24:43,960 --> 00:24:45,760
THAT THERE IS AT LEAST ONE DRUG

743
00:24:45,760 --> 00:24:48,160
IN THE COCKTAIL THAT HAS AN AUC

744
00:24:48,160 --> 00:24:51,560
VALUE OF LESS THAN .55, THE

745
00:24:51,560 --> 00:24:52,240
PAYABLE EITHER HAD STABLE

746
00:24:52,240 --> 00:24:53,480
DISEASE OR PARTIAL RESPONSE.

747
00:24:53,480 --> 00:24:56,240
WHEREAS IF THE AUC VALUE IS

748
00:24:56,240 --> 00:24:58,440
HIGHER THAN THAT, THEN THE

749
00:24:58,440 --> 00:24:59,440
PATIENT PROGRESSED.

750
00:24:59,440 --> 00:25:01,720
WE REALIZED THAT IDENTIFYING A

751
00:25:01,720 --> 00:25:05,880
DRUG THAT HAS AN AUC VALUELESS

752
00:25:05,880 --> 00:25:07,600
THAN .55 IN THE ORGANOID CULTURE

753
00:25:07,600 --> 00:25:12,480
GIVES US ABOUT 80 TO 85%

754
00:25:12,480 --> 00:25:13,480
PREDICTIVE POWER TO TELL WHETHER

755
00:25:13,480 --> 00:25:16,640
THE DRUG IS GOING TO WORK OR

756
00:25:16,640 --> 00:25:17,560
NOT.

757
00:25:17,560 --> 00:25:19,600
THE SECOND INSIGHT WE GAINED, IN

758
00:25:19,600 --> 00:25:23,680
A DRUG COCKTAIL, PRESENCE WITH

759
00:25:23,680 --> 00:25:25,880
AUC -- WAS SUFFICIENT TO PROVIDE

760
00:25:25,880 --> 00:25:26,400
POSITIVE RESPONSE.

761
00:25:26,400 --> 00:25:29,200
SO THIS HAS SEVERAL

762
00:25:29,200 --> 00:25:30,960
IMPLICATIONS.

763
00:25:30,960 --> 00:25:32,640
THERE'S A COUNTERINTUITIVE

764
00:25:32,640 --> 00:25:34,080
APPLICATION WHICH IS IDENTIFY

765
00:25:34,080 --> 00:25:35,440
THE DRUGS THAT DO NOT WORK IN

766
00:25:35,440 --> 00:25:37,680
THE ORGANOID, YOU CAN EXCLUDE

767
00:25:37,680 --> 00:25:40,800
THEM IN THE CLINIC AND YOU MAY

768
00:25:40,800 --> 00:25:43,760
ACTUALLY EXPERIENCE THAT AS

769
00:25:43,760 --> 00:25:44,280
WELL.

770
00:25:44,280 --> 00:25:45,920
PERHAPS WE CAN USE THEM AS A WAY

771
00:25:45,920 --> 00:25:49,080
TO UNDERSTAND WHY PATIENT TO

772
00:25:49,080 --> 00:25:50,000
PATIENT DIFFERENCES IN DRUG

773
00:25:50,000 --> 00:25:50,920
TERRITORIMENT DO OCCUR.

774
00:25:50,920 --> 00:25:51,960
SO I'M HOPING THESE ARE THE KIND

775
00:25:51,960 --> 00:25:54,080
OF QUESTIONS WE WILL BE ABLE TO

776
00:25:54,080 --> 00:25:58,400
ANSWER IN OUR LAB HERE IN LCBG

777
00:25:58,400 --> 00:25:59,440
IN COLLABORATION WITH THE

778
00:25:59,440 --> 00:26:04,960
CLINICAL TEAM HERE.

779
00:26:04,960 --> 00:26:06,480
SO THE LAST RESULTS I WANT TO

780
00:26:06,480 --> 00:26:07,760
SUMMARIZE IS HOW WE HAVE USED

781
00:26:07,760 --> 00:26:09,040
ORGANOIDS IN THE CONTEXT OF

782
00:26:09,040 --> 00:26:10,920
DISCOVERING T-CELL RECEPTORS,

783
00:26:10,920 --> 00:26:12,480
TUMOR TARGETING T-CELL

784
00:26:12,480 --> 00:26:13,040
RECEPTORS.

785
00:26:13,040 --> 00:26:14,080
IN THIS CONTEXT, WE ACTUALLY

786
00:26:14,080 --> 00:26:15,960
TOOK A COMPLETELY DIFFERENT

787
00:26:15,960 --> 00:26:17,320
APPROACH, AND WE DECIDED NOT TO

788
00:26:17,320 --> 00:26:19,960
FOCUS ON TUMOR INFILTRATING

789
00:26:19,960 --> 00:26:22,280
LYMPHOCYTES BUT REALLY TO FOCUS

790
00:26:22,280 --> 00:26:23,560
ON PERIPHERAL BLOOD DERIVED

791
00:26:23,560 --> 00:26:23,920
T-CELLS.

792
00:26:23,920 --> 00:26:25,560
AND THE REASON IS WE THINK THIS

793
00:26:25,560 --> 00:26:27,240
MAY ALLOW US TO FIND T-CELL

794
00:26:27,240 --> 00:26:29,960
RECEPTORS THAT ARE DIFFICULT TO

795
00:26:29,960 --> 00:26:32,360
FIND BECAUSE FINDING T-CELL

796
00:26:32,360 --> 00:26:35,000
RECEPTORS IN THE PERIPHERAL

797
00:26:35,000 --> 00:26:37,240
BLOOD IS USUALLY LIKE FINDING A

798
00:26:37,240 --> 00:26:38,320
NEEDLE IN A HAYSTACK.

799
00:26:38,320 --> 00:26:40,400
WE DON'T WANT BIAS TOWARDS ANY

800
00:26:40,400 --> 00:26:41,840
ANTIGENS OR NEOANTIGENS, WE JUST

801
00:26:41,840 --> 00:26:43,240
WANT NATURE TO SELECT ITSELF.

802
00:26:43,240 --> 00:26:44,680
SO THE WAY IT'S DONE IS WE

803
00:26:44,680 --> 00:26:45,520
GENERATE ORGANOID FROM A

804
00:26:45,520 --> 00:26:46,200
PATIENT, AT THE SAME TIME WE

805
00:26:46,200 --> 00:26:48,400
ALSO GET PERIPHERAL BLOOD, WE

806
00:26:48,400 --> 00:26:49,080
EXPAND THE T-CELLS FROM THE

807
00:26:49,080 --> 00:26:53,440
PATIENT AND WE CO-CULTURE THEM

808
00:26:53,440 --> 00:26:54,720
TOGETHER, TWO WEEKS, A SELECTIVE

809
00:26:54,720 --> 00:26:57,360
NUMBER OF T-CELLS WILL NOW

810
00:26:57,360 --> 00:26:59,400
EXPAND, ORGANOID PRIMED T-CELLS.

811
00:26:59,400 --> 00:27:00,840
NOW THE BIOLOGY WORKS

812
00:27:00,840 --> 00:27:02,680
BEAUTIFULLY SO HERE IS AN

813
00:27:02,680 --> 00:27:04,400
ORGANOID AND A PBMC DIRECT

814
00:27:04,400 --> 00:27:05,640
T-CELL FROM THE SAME PATIENT.

815
00:27:05,640 --> 00:27:07,640
YOU PUT IT TO THE AND THEY START

816
00:27:07,640 --> 00:27:09,040
KILLING AND ATTACKING THE

817
00:27:09,040 --> 00:27:10,920
ORGANOID AND IN A PERIOD OF 14

818
00:27:10,920 --> 00:27:11,680
DAYS, THEY COMPLETELY KILL ALL

819
00:27:11,680 --> 00:27:13,840
THE TUMORS, AND IN THE PROCESS,

820
00:27:13,840 --> 00:27:15,200
THE T-CELLS GET ACTIVATED AND

821
00:27:15,200 --> 00:27:16,240
EXPAND IN NUMBER.

822
00:27:16,240 --> 00:27:20,000
ONCE YOU GENERATE A POPULATION

823
00:27:20,000 --> 00:27:22,400
OF CELLS, THEY'RE REALLY ROBUST

824
00:27:22,400 --> 00:27:23,520
IN KILLING THE TUMOR.

825
00:27:23,520 --> 00:27:27,120
IF YOU CHALLENGED -- THEY

826
00:27:27,120 --> 00:27:28,360
ACTUALLY WILL KILL IT WITHIN 10

827
00:27:28,360 --> 00:27:28,560
HOURS.

828
00:27:28,560 --> 00:27:30,720
WHEN YOU PUT THEM TOGETHER, THEY

829
00:27:30,720 --> 00:27:32,760
COMPLETELY OBLITERATE THE

830
00:27:32,760 --> 00:27:34,440
ORGANORGANIZE QUITE EFFICIENTLYD

831
00:27:34,440 --> 00:27:35,560
THAT'S HOW THEY SEEM TO WORK IN

832
00:27:35,560 --> 00:27:38,040
ALL THE CASES WE'VE TESTED SO

833
00:27:38,040 --> 00:27:39,080
FAR.

834
00:27:39,080 --> 00:27:41,560
NOW THESE T-CELLS HAVE ALL THE

835
00:27:41,560 --> 00:27:42,880
CYTOTOXIC T-CELL SIGNATURES IN

836
00:27:42,880 --> 00:27:44,200
RESPONSE TO STIMULATION OR

837
00:27:44,200 --> 00:27:46,400
EXPOSURE TO ORGANOIDS, THEY

838
00:27:46,400 --> 00:27:47,840
UNDERGO PROLIFERATION, THEY

839
00:27:47,840 --> 00:27:51,000
SECRETE IN DIFFERENT -- IT

840
00:27:51,000 --> 00:27:52,160
REALLY TOLD US WE CAN NOW

841
00:27:52,160 --> 00:27:54,560
ESTABLISH A POPULATION OF

842
00:27:54,560 --> 00:27:56,760
T-CELLS BY CO-CULTURING WITH

843
00:27:56,760 --> 00:27:57,280
ORGANOIDS.

844
00:27:57,280 --> 00:27:58,960
SO THE QUESTION IS, IS THERE

845
00:27:58,960 --> 00:28:00,360
CLONAL EXPANSION GOING ON,

846
00:28:00,360 --> 00:28:01,560
BECAUSE THAT IS CENTRAL FOR US

847
00:28:01,560 --> 00:28:03,440
TO REALLY BE ABLE TO TAKE TO THE

848
00:28:03,440 --> 00:28:03,920
NEXT LEVEL.

849
00:28:03,920 --> 00:28:07,040
IN ORDER TO DO THAT, WE TOOK

850
00:28:07,040 --> 00:28:09,680
MATCHED OBMC AND OP T-CELLS FROM

851
00:28:09,680 --> 00:28:12,760
THE SAME PATIENT, THEN WE ASKED

852
00:28:12,760 --> 00:28:14,720
WHAT FRACTION OF THE TCR YOU

853
00:28:14,720 --> 00:28:16,720
FIND IS REPRESENTATIVE OF THE

854
00:28:16,720 --> 00:28:17,640
ENTIRE POPULATION OF THAT

855
00:28:17,640 --> 00:28:18,240
CULTURE.

856
00:28:18,240 --> 00:28:19,320
THE RESULTS WERE QUITE

857
00:28:19,320 --> 00:28:21,040
FASCINATING AND SURPRISING.

858
00:28:21,040 --> 00:28:22,720
FOR THE PBMCs JUST AS

859
00:28:22,720 --> 00:28:26,800
EVERYBODY WOULD EXPECT, MOST

860
00:28:26,800 --> 00:28:28,240
TCRs WERE PRESENT IN .1%.

861
00:28:28,240 --> 00:28:32,280
THERE WERE SOME TCRs THAT WERE

862
00:28:32,280 --> 00:28:34,320
5 TO 6% BUT MOST OF THEM VERY

863
00:28:34,320 --> 00:28:35,240
LOW IN NUMBER.

864
00:28:35,240 --> 00:28:40,640
BY CONTRAST, THE OP T-CELLS, --

865
00:28:40,640 --> 00:28:42,760
SOMETIMES ONE TCR WAS ABOUT 70

866
00:28:42,760 --> 00:28:43,720
TO 90%.

867
00:28:43,720 --> 00:28:45,360
THAT TOLD US THAT THERE IS

868
00:28:45,360 --> 00:28:48,960
SEVERAL ORDERS OF MAGNITUDE OF

869
00:28:48,960 --> 00:28:50,160
EXPANSION HAPPENING IN THIS

870
00:28:50,160 --> 00:28:52,040
CO-CULTURE SETTING, EMPOWERING

871
00:28:52,040 --> 00:28:53,120
US TO THINK PERHAPS THIS IS A

872
00:28:53,120 --> 00:28:55,280
WAY WE CAN FISH OUT THE T-CELL

873
00:28:55,280 --> 00:28:56,400
RECEPTORS THAT ARE ABLE TO

874
00:28:56,400 --> 00:28:57,600
TARGET THESE TUMOR CELLS FROM

875
00:28:57,600 --> 00:28:57,960
THE CIRCULATION.

876
00:28:57,960 --> 00:29:01,040
SO WE WANTED TO KNOW IF THESE

877
00:29:01,040 --> 00:29:01,880
TCRs ARE ACTUALLY THE ONES

878
00:29:01,880 --> 00:29:03,280
THAT ARE DRIVING THE PHENOTYPE,

879
00:29:03,280 --> 00:29:05,400
AND IF THEY ARE TRANSFERABLE TO

880
00:29:05,400 --> 00:29:05,880
OTHER CELLS.

881
00:29:05,880 --> 00:29:09,080
TO DO THAT, WE SEQUENCE THE

882
00:29:09,080 --> 00:29:11,280
VARIABLE CHAINS AND THEN CREATED

883
00:29:11,280 --> 00:29:13,480
A CHIMERIC TCR EXPRESSING THE

884
00:29:13,480 --> 00:29:14,480
CONSTANT REGION FOR MOUSE.

885
00:29:14,480 --> 00:29:17,560
NOW YOU CAN EXPRESS THIS WITH

886
00:29:17,560 --> 00:29:19,400
THIS KIE ME TICK RECEPTOR AND

887
00:29:19,400 --> 00:29:22,200
THEN ASK WILL THIS T-CELL HAVE

888
00:29:22,200 --> 00:29:23,680
THE ABILITY TO ATTACK AND KILL

889
00:29:23,680 --> 00:29:26,840
THE ORGANOID THAT THE T-CELL

890
00:29:26,840 --> 00:29:28,040
RECEPTOR WAS GENERATED FROM?

891
00:29:28,040 --> 00:29:28,920
AND THE ANSWER IS YES.

892
00:29:28,920 --> 00:29:32,320
SO WE TOOK A PATIENT 10 DERIVED

893
00:29:32,320 --> 00:29:33,520
T-CELL RECEPTOR SO THAT'S

894
00:29:33,520 --> 00:29:34,960
AUTOLOGOUS, THAT SHOULD KILL,

895
00:29:34,960 --> 00:29:37,160
WHEREAS A PERSON 38 ORGANOID

896
00:29:37,160 --> 00:29:39,240
WHICH IS ALLOGENEIC SO WE DON'T

897
00:29:39,240 --> 00:29:40,520
KNOW WHETHER IT WILL KILL OR

898
00:29:40,520 --> 00:29:41,240
NOT.

899
00:29:41,240 --> 00:29:44,320
SO WHEN YOU CO-CULTURE THAT,

900
00:29:44,320 --> 00:29:48,120
WHEN -- IT QUITE EFFECTIVELY

901
00:29:48,120 --> 00:29:53,000
KILLS WITHIN 24 HOURS AND 38 ON

902
00:29:53,000 --> 00:29:54,400
THE OTHER HAND IS QUITE WEEK.

903
00:29:54,400 --> 00:29:56,880
THIS KILLING IS STILL MEDIATED

904
00:29:56,880 --> 00:29:58,640
BY ANTIGEN PRESENTATION BY THE

905
00:29:58,640 --> 00:29:59,840
TUMOR CELLS BECAUSE IF YOU THROW

906
00:29:59,840 --> 00:30:02,560
IN A FUNCTION BLOCKING ANTIBODY,

907
00:30:02,560 --> 00:30:03,880
YOU COMPLETELY BLOCK THIS

908
00:30:03,880 --> 00:30:04,680
PHENOTYPE.

909
00:30:04,680 --> 00:30:07,280
SO THESE ENGINEERED T-CELLS SEE

910
00:30:07,280 --> 00:30:08,040
SOMETHING, WE DON'T KNOW WHAT

911
00:30:08,040 --> 00:30:10,520
THAT IS, ON THE TUMOR CELL, AND

912
00:30:10,520 --> 00:30:11,800
THEN KILLS THE TUMOR CELLS.

913
00:30:11,800 --> 00:30:13,440
SO THIS HAS REALLY ALLOWED US

914
00:30:13,440 --> 00:30:15,400
INTO A NEW ERA OF THINKING THAT

915
00:30:15,400 --> 00:30:16,280
PERHAPS THIS WAY OF DOING THIS

916
00:30:16,280 --> 00:30:20,720
CAN BE ONE WAY OF DOING ADAPTIVE

917
00:30:20,720 --> 00:30:23,680
CELL THERAPY, WHICH IS

918
00:30:23,680 --> 00:30:25,960
LABORIOUS, MAYBE TI TIME-CONSUM,

919
00:30:25,960 --> 00:30:27,000
BUT ON THE OTHER HAND USING THIS

920
00:30:27,000 --> 00:30:29,360
PLATFORM TO IDENTIFY TUMOR

921
00:30:29,360 --> 00:30:31,960
TARGETING TCRs IS QUITE A

922
00:30:31,960 --> 00:30:34,800
POWERFUL STRATEGY AND THAT'S

923
00:30:34,800 --> 00:30:36,120
SOMETHING WE'RE GOING TO HEAVILY

924
00:30:36,120 --> 00:30:37,760
INVEST ON, SO HOPING TO BE ABLE

925
00:30:37,760 --> 00:30:39,960
TO DO A LOT OF THESE THINGS HERE

926
00:30:39,960 --> 00:30:40,840
AND AT NCI.

927
00:30:40,840 --> 00:30:43,120
SO BEYOND THESE THREE STORIES,

928
00:30:43,120 --> 00:30:45,520
WE HAVE USED ORGANOID PLATFORM

929
00:30:45,520 --> 00:30:47,160
FOR OTHER MULTIPLE ORGANIZATIONS

930
00:30:47,160 --> 00:30:50,120
BUT I DON'T WANT TO -- WE

931
00:30:50,120 --> 00:30:51,000
PUBLISHED MOST OF THEM AND

932
00:30:51,000 --> 00:30:52,200
YOU'LL BE ABLE TO FIND THEM.

933
00:30:52,200 --> 00:30:53,880
SO GIVEN THE POWER OF THIS, WHEN

934
00:30:53,880 --> 00:30:55,520
I MOVED HERE WITH THE SUPPORT OF

935
00:30:55,520 --> 00:30:58,600
THOMAS TELLY AND CATHY KELLY,

936
00:30:58,600 --> 00:31:02,440
CATHY AND I FACILITATED -- ONE

937
00:31:02,440 --> 00:31:04,240
IS TO GENERATE ORGANOID MODELS

938
00:31:04,240 --> 00:31:05,240
BY COLLABORATING WITH THE

939
00:31:05,240 --> 00:31:06,800
CLINICAL TEAM HERE AT NCI FROM

940
00:31:06,800 --> 00:31:08,600
THE PATIENTS WHO UNDERGO SURGERY

941
00:31:08,600 --> 00:31:10,040
HERE, AND POTENTIALLY MAKE THEM

942
00:31:10,040 --> 00:31:11,520
AS A RESOURCE FOR THE COMMUNITY

943
00:31:11,520 --> 00:31:12,400
HERE.

944
00:31:12,400 --> 00:31:14,800
THE SECOND IS TO SERVE AS A

945
00:31:14,800 --> 00:31:16,480
TRAINING AND KNOWLEDGE BASE, TO

946
00:31:16,480 --> 00:31:17,960
REALLY PROVIDE HANDS-ON TRAINING

947
00:31:17,960 --> 00:31:19,400
FOR THOSE WHO MAY BE NEEDING

948
00:31:19,400 --> 00:31:20,360
TRAINING AND MENTOR THEM THROUGH

949
00:31:20,360 --> 00:31:21,600
THE TRANSFER OF KNOWLEDGE TO

950
00:31:21,600 --> 00:31:22,560
THEIR OWN SPACE.

951
00:31:22,560 --> 00:31:24,440
AND WHATEVER MEDIA WE HAVE

952
00:31:24,440 --> 00:31:27,080
GENERATED FOR SOME ORGANOIDS,

953
00:31:27,080 --> 00:31:29,520
WE'LL PROVIDE -- THE RECIPES,

954
00:31:29,520 --> 00:31:30,600
WILL PROVIDE THEM TO THE LAB WHO

955
00:31:30,600 --> 00:31:32,240
MAY BE INTERESTED IN USING THEM.

956
00:31:32,240 --> 00:31:33,440
SO I'M HOPING THIS WILL TAKE

957
00:31:33,440 --> 00:31:34,880
OFF -- IT'S ALREADY TAKING OFF,

958
00:31:34,880 --> 00:31:36,400
ACTUALLY, THANKS TO THE CLINICAL

959
00:31:36,400 --> 00:31:37,560
TEAM, JONATHAN IS SITTING THERE,

960
00:31:37,560 --> 00:31:39,160
WHERE WE GET A LOT OF SAMPLES

961
00:31:39,160 --> 00:31:40,600
AND WE'RE GENERATING ORGANOIDS

962
00:31:40,600 --> 00:31:42,640
ON A ROUTINE BASIS.

963
00:31:42,640 --> 00:31:44,680
SO NOW I'M GOING TO SHIFT GEAR

964
00:31:44,680 --> 00:31:46,200
AND GO INTO THE SECOND TOPIC OF

965
00:31:46,200 --> 00:31:48,480
HOW WE ARE REALLY INVESTIGATING

966
00:31:48,480 --> 00:31:51,200
THE ROLE PLAYED BY CELL POLARITY

967
00:31:51,200 --> 00:31:53,760
PROTEINS IN THE CONTEXT OF DRUG

968
00:31:53,760 --> 00:31:55,240
RESISTANCE.

969
00:31:55,240 --> 00:31:57,040
SO I'M NOT SURE HOW MANY OF YOU

970
00:31:57,040 --> 00:31:58,400
KNOW OR HEARD TOO MUCH ABOUT

971
00:31:58,400 --> 00:31:59,360
CELL POLARITY BUT FOR THE SAKE

972
00:31:59,360 --> 00:32:01,200
OF THE TRAINEES IN THE AUDIENCE,

973
00:32:01,200 --> 00:32:04,200
THE CELL POLARITY IS ACTUALLY A

974
00:32:04,200 --> 00:32:05,920
FUNDAMENTAL PROPERTY OF ALL

975
00:32:05,920 --> 00:32:06,520
CELLS.

976
00:32:06,520 --> 00:32:09,040
IT IS THE MECHANISM BY WHICH

977
00:32:09,040 --> 00:32:10,200
ASYMMETRY IS CREATED WITHIN A

978
00:32:10,200 --> 00:32:10,400
CELL.

979
00:32:10,400 --> 00:32:11,520
THE FRONT OF A CELL LOOKS

980
00:32:11,520 --> 00:32:12,240
DIFFERENT FROM THE REAR OF A

981
00:32:12,240 --> 00:32:14,440
SELL IN A MIGRATING CELL.

982
00:32:14,440 --> 00:32:15,720
THE PORTION OF THE CELL THAT

983
00:32:15,720 --> 00:32:18,200
FACES THE LUMINAL SPACE OF THE

984
00:32:18,200 --> 00:32:19,280
EMPTY DUCTAL SPACE IS DIFFERENT

985
00:32:19,280 --> 00:32:20,920
FROM WHAT IS IN THE BASE.

986
00:32:20,920 --> 00:32:22,520
ALL THAT ASYMMETRY IS CREATED BY

987
00:32:22,520 --> 00:32:23,680
THE CELL POLARITY PROTEINS.

988
00:32:23,680 --> 00:32:25,920
NOW THIS POLARITY IS A

989
00:32:25,920 --> 00:32:28,400
FUNDAMENTALLY IMPORTANT FOR

990
00:32:28,400 --> 00:32:28,760
PHYSIOLOGY.

991
00:32:28,760 --> 00:32:31,200
POLARITY DICTATES HOW YOU ABSORB

992
00:32:31,200 --> 00:32:33,080
NUTRIENTS IN THE GUT AND YOU

993
00:32:33,080 --> 00:32:35,040
SECRETE MILK IN THE DUCT, IN THE

994
00:32:35,040 --> 00:32:35,640
MAMMARY GLAND.

995
00:32:35,640 --> 00:32:37,520
THOSE ARE ALL PROCESSES STRICTLY

996
00:32:37,520 --> 00:32:39,000
REGULATED BY THESE PROTEINS.

997
00:32:39,000 --> 00:32:41,600
SO DISRUPTION OF THIS POLARITY

998
00:32:41,600 --> 00:32:43,760
OF PROTEINS ALMOST ALWAYS LEADS

999
00:32:43,760 --> 00:32:47,240
TO THE DIRECTIONAL PROCESSES

1000
00:32:47,240 --> 00:32:48,320
WHICH LEAD TO DISEASE.

1001
00:32:48,320 --> 00:32:49,320
NOW THERE'S BEEN A LOT OF

1002
00:32:49,320 --> 00:32:50,680
STUDIES IN MODEL ORGANISMS

1003
00:32:50,680 --> 00:32:53,840
TRYING TO UNDERSTAND THIS,

1004
00:32:53,840 --> 00:32:56,880
MOSTLY FROM DROSOPHILA C.

1005
00:32:56,880 --> 00:32:58,400
ELEGANS -- POLARITY PROTEINS,

1006
00:32:58,400 --> 00:33:00,640
THEY'RE SCAFFOLDING MOLECULES

1007
00:33:00,640 --> 00:33:01,600
AND SOME KINASES.

1008
00:33:01,600 --> 00:33:03,080
I WON'T GO INTO THE DETAILS OF

1009
00:33:03,080 --> 00:33:05,000
ALL THESE MOLECULES AT THIS

1010
00:33:05,000 --> 00:33:05,240
POINT.

1011
00:33:05,240 --> 00:33:07,000
THE REASON I GOT INTERESTED IN

1012
00:33:07,000 --> 00:33:10,240
POLARITY PROTEINS A LONG TIME

1013
00:33:10,240 --> 00:33:13,160
AGO IS SOME OF THE STUDIES

1014
00:33:13,160 --> 00:33:14,600
PUBLISHED IN DROSOPHILA.

1015
00:33:14,600 --> 00:33:15,880
IF YOU HAVE A LOSS OF FUNCTION

1016
00:33:15,880 --> 00:33:17,360
MUTATION OF A POLARITY PROTEIN,

1017
00:33:17,360 --> 00:33:19,280
NOT ONLY DO YOU LOSE CELL

1018
00:33:19,280 --> 00:33:22,240
STRUCTURE, BUT THE CELL GAINS

1019
00:33:22,240 --> 00:33:22,920
UNCONTROLLABLE GROWTH ADVANTAGE.

1020
00:33:22,920 --> 00:33:25,360
IT ACTUALLY GROWS TO FORM A

1021
00:33:25,360 --> 00:33:28,440
TUMOR-LIKE PHENOTYPE IN -- AND

1022
00:33:28,440 --> 00:33:30,080
THESE FLIES ARE IN THE LARVAE.

1023
00:33:30,080 --> 00:33:32,880
SO LED TO GENETICIST TO COIN THE

1024
00:33:32,880 --> 00:33:34,960
TERM THAT THESE ARE NEOPLASTIC

1025
00:33:34,960 --> 00:33:35,800
TUMOR SUPPRESSORS.

1026
00:33:35,800 --> 00:33:36,880
BUT IT WAS NOT KNOWN AT THAT

1027
00:33:36,880 --> 00:33:38,720
TIME WHETHER THEY ARE TUMOR

1028
00:33:38,720 --> 00:33:40,040
SUPPRESSORS IN MAMMALIAN CELLS

1029
00:33:40,040 --> 00:33:40,720
OR NOT.

1030
00:33:40,720 --> 00:33:42,040
SO THAT'S HOW WE STARTED IN MY

1031
00:33:42,040 --> 00:33:44,200
LAB ASKING THE QUESTION, ARE

1032
00:33:44,200 --> 00:33:46,200
THESE TUMOR SUPPRESSORS IN

1033
00:33:46,200 --> 00:33:47,760
MAMMALIAN CONTEXT?

1034
00:33:47,760 --> 00:33:51,920
HE PUBLISHES -- LOSS OF FUNCTION

1035
00:33:51,920 --> 00:33:53,880
OF POLARITY PROTEINS WILL

1036
00:33:53,880 --> 00:33:56,520
COOPERATE WITH ONCOGENES TO

1037
00:33:56,520 --> 00:33:59,800
INDUCE TUMOR, TRANSFORM CELLS TO

1038
00:33:59,800 --> 00:34:01,440
INDUCE METASTATIC PHENOTYPE AND

1039
00:34:01,440 --> 00:34:02,080
SO FORTH.

1040
00:34:02,080 --> 00:34:06,920
ALONG THE WAY WE MADE SOME

1041
00:34:06,920 --> 00:34:08,720
POLARITY PROTEINS THAT, WHEN

1042
00:34:08,720 --> 00:34:10,560
OVEREXPRESSED, ACTUALLY PROMOTE

1043
00:34:10,560 --> 00:34:11,320
PROLIFERATION.

1044
00:34:11,320 --> 00:34:12,520
SOME MIGHT SHOW THAT EVEN IF YOU

1045
00:34:12,520 --> 00:34:14,320
MAKE A TRANSGENIC MOUSE MODEL,

1046
00:34:14,320 --> 00:34:16,360
WE CAN INDUCE GENERATION OF

1047
00:34:16,360 --> 00:34:16,600
TUMORS.

1048
00:34:16,600 --> 00:34:18,920
SO THIS REALLY PUZZLED US FOR A

1049
00:34:18,920 --> 00:34:20,520
WHILE AND ALSO WHEN YOU LOOK AT

1050
00:34:20,520 --> 00:34:22,280
THE GENOMICS DATABASE, THERE WAS

1051
00:34:22,280 --> 00:34:23,600
ACTUALLY A CLASS OF POLARITY

1052
00:34:23,600 --> 00:34:24,840
PROTEINS, A GROUP OF THEM, A FEW

1053
00:34:24,840 --> 00:34:26,040
THAT ARE FREQUENTLY

1054
00:34:26,040 --> 00:34:28,000
OVEREXPRESSED AND AMPLIFIED IN

1055
00:34:28,000 --> 00:34:28,440
MULTIPLE CANCER TYPES.

1056
00:34:28,440 --> 00:34:30,200
SO THIS MADE US THINK THAT MAYBE

1057
00:34:30,200 --> 00:34:31,840
NOT ALL POLARITY PROTEINS ARE

1058
00:34:31,840 --> 00:34:33,880
THE SAME, MAYBE SOME OF THES ARE

1059
00:34:33,880 --> 00:34:35,080
POSITIVE REGULATORS OF BIOLOGY.

1060
00:34:35,080 --> 00:34:37,320
AND THE REASON -- WE WANTED TO

1061
00:34:37,320 --> 00:34:38,640
REALLY PURSUE THIS BECAUSE IF

1062
00:34:38,640 --> 00:34:42,480
THEY ARE POSITIVE REGULATORS,

1063
00:34:42,480 --> 00:34:43,840
THEY MAY USE A NEW CLASS OF

1064
00:34:43,840 --> 00:34:46,480
TARGETS THAT YOU CAN GO AFTER,

1065
00:34:46,480 --> 00:34:48,000
IT'S NOT A DNA DAMAGE REPAIR OR

1066
00:34:48,000 --> 00:34:52,400
ANY OF THOSE PROCESSES THAT ARE

1067
00:34:52,400 --> 00:34:53,280
ROUTINE STUDIES THERAPEUTIC

1068
00:34:53,280 --> 00:34:53,560
ADVANTAGE.

1069
00:34:53,560 --> 00:34:56,800
WITH THAT IN MIND WE INITIATED A

1070
00:34:56,800 --> 00:35:00,200
PROGRAM IN THE LAB TO ASK --

1071
00:35:00,200 --> 00:35:01,520
PROMOTERS OF CANCER BIOLOGY.

1072
00:35:01,520 --> 00:35:04,400
THE GENE WE FOCUSED ON WAS

1073
00:35:04,400 --> 00:35:07,040
CALLED LETHAL GIANT LARVAE,

1074
00:35:07,040 --> 00:35:09,520
RECENTLY DISCOVERED IN

1075
00:35:09,520 --> 00:35:10,080
DROSOPHILA.

1076
00:35:10,080 --> 00:35:13,360
LOSS OF MUTATION RESULTS IN

1077
00:35:13,360 --> 00:35:17,880
LETHAL GIANT LARVAE BECAUSE --

1078
00:35:17,880 --> 00:35:19,760
GROW OUT OF CONTROL.

1079
00:35:19,760 --> 00:35:21,960
AT THAT TIME THE LITERATURE WAS

1080
00:35:21,960 --> 00:35:24,400
REALLY VAGUE IN IS IT REALLY A

1081
00:35:24,400 --> 00:35:26,240
TUMOR SUPPRESSOR OR PROMOTER.

1082
00:35:26,240 --> 00:35:29,960
THERE WERE RESULTS THAT WERE

1083
00:35:29,960 --> 00:35:31,000
PUBLISHED, BOTH CAN GO IN BOTH

1084
00:35:31,000 --> 00:35:31,480
DIRECTIONS.

1085
00:35:31,480 --> 00:35:33,360
SO BECAUSE OF THAT, WE ACTUALLY

1086
00:35:33,360 --> 00:35:37,880
TOOK THIS MOLECULE AND YOU ALSO

1087
00:35:37,880 --> 00:35:39,800
SAY TO -- IT WAS SO COMPLICATED

1088
00:35:39,800 --> 00:35:41,840
THAT IT WAS CALLED THE TAR BABY,

1089
00:35:41,840 --> 00:35:43,720
BECAUSE ANYBODY THAT TOUCHES LGL

1090
00:35:43,720 --> 00:35:45,120
CANNOT REALLY COMPLETE THEIR

1091
00:35:45,120 --> 00:35:47,280
STUDY BECAUSE IT GOES BOTH WAYS,

1092
00:35:47,280 --> 00:35:48,520
DEPENDING ON THE TOPIC, SO

1093
00:35:48,520 --> 00:35:51,680
NOBODY COULD REALLY UNDERSTAND

1094
00:35:51,680 --> 00:35:54,200
HOW LGL WAS WORKING WELL AT THAT

1095
00:35:54,200 --> 00:35:54,840
TIME.

1096
00:35:54,840 --> 00:35:57,600
SO SATO NOW HAS HIS OWN LAB IN

1097
00:35:57,600 --> 00:36:00,920
JAPAN AND LOOKED AT LLGL EGGS

1098
00:36:00,920 --> 00:36:03,200
PRETION IN CANCER AS A FUNCTION

1099
00:36:03,200 --> 00:36:04,800
OF CLINICAL OUTCOME.

1100
00:36:04,800 --> 00:36:10,120
IF HE LOOKS, HIGH EXPRESSION

1101
00:36:10,120 --> 00:36:16,800
CORRELATED WITH GOOD OUTCOME IN

1102
00:36:16,800 --> 00:36:19,200
ERPR POSITIVE CANCERS.

1103
00:36:19,200 --> 00:36:20,480
THIS IS CONSISTENT WITH THE

1104
00:36:20,480 --> 00:36:22,120
REASONING THAT LLGL1 MAY BE A

1105
00:36:22,120 --> 00:36:23,320
ACTUAL MORE SUPPRESSOR, SO HIGH

1106
00:36:23,320 --> 00:36:24,040
IS GOOD.

1107
00:36:24,040 --> 00:36:25,160
THE RESULTS WERE OPPOSITE WHEN

1108
00:36:25,160 --> 00:36:28,200
YOU LOOKED AT LGL2.

1109
00:36:28,200 --> 00:36:31,160
WHEN YOU SEE HIGHER LEVELS OF

1110
00:36:31,160 --> 00:36:33,240
LGL2, IT HAD A STRONG

1111
00:36:33,240 --> 00:36:34,640
CORRELATION WITH POOR OUTCOME.

1112
00:36:34,640 --> 00:36:36,280
SO YOU'LL SEE AT THE RED LINE

1113
00:36:36,280 --> 00:36:38,280
HERE WHICH IS HIGH, THE PATIENTS

1114
00:36:38,280 --> 00:36:40,160
DIE FASTER COMPARED TO THE BLACK

1115
00:36:40,160 --> 00:36:40,400
ONE.

1116
00:36:40,400 --> 00:36:44,280
AND THIS BIOLOGY WAS MORE

1117
00:36:44,280 --> 00:36:50,040
SPECIFIC TO ERPR POSITIVE, NOT

1118
00:36:50,040 --> 00:36:50,600
NEGATIVE.

1119
00:36:50,600 --> 00:36:52,760
SO YOU TOOK A PANEL AND STARTED

1120
00:36:52,760 --> 00:37:00,760
LOOKING FOR LGL1 AND  LGL2.

1121
00:37:00,760 --> 00:37:04,320
LGL2 EXPRESSION WAS HIGHER IN  

1122
00:37:04,320 --> 00:37:05,800
ERP POSITIVE COMPARED TO

1123
00:37:05,800 --> 00:37:09,040
NEGATIVE.

1124
00:37:09,040 --> 00:37:10,800
LGL1 IS MORE NEGATIVE THAN ALL

1125
00:37:10,800 --> 00:37:14,040
THAT HE TSH THERE'S A SELECTION

1126
00:37:14,040 --> 00:37:16,520
FOR HIGHER LEVELS OF LGL2 IN

1127
00:37:16,520 --> 00:37:18,040
THESE CELLS.

1128
00:37:18,040 --> 00:37:20,560
TO TEST IT, HE KNOCKED DOWN --

1129
00:37:20,560 --> 00:37:23,200
I'M SORRY -- WHAT HE DID WAS HE

1130
00:37:23,200 --> 00:37:26,720
ACTUALLY GENERATED MCF 7 CELLS

1131
00:37:26,720 --> 00:37:29,000
THAT OVER -- AND ASK DO THEY

1132
00:37:29,000 --> 00:37:30,320
GIVE ANY SORT OF PROLIFERATIVE

1133
00:37:30,320 --> 00:37:30,600
ADVANTAGE.

1134
00:37:30,600 --> 00:37:31,800
THE RESULTS WERE CONFUSING FOR A

1135
00:37:31,800 --> 00:37:32,320
LONG TIME.

1136
00:37:32,320 --> 00:37:33,920
HE COULD NOT FIND ANY BIOLOGY

1137
00:37:33,920 --> 00:37:37,040
THAT WAS ACTUALLY SUGGESTING

1138
00:37:37,040 --> 00:37:39,040
THAT LGL1 AND L

1139
00:37:39,040 --> 00:37:39,880
GL2 OVEREXPRESSION WAS ACTUALLY

1140
00:37:39,880 --> 00:37:41,440
HELPFUL FOR THESE CELLS.

1141
00:37:41,440 --> 00:37:42,840
UNTIL HE CHALLENGED THESE CELLS

1142
00:37:42,840 --> 00:37:44,400
WITH THE NUTRIENT STRESS

1143
00:37:44,400 --> 00:37:45,160
CONDITIONS.

1144
00:37:45,160 --> 00:37:50,000
HE HE SWITCHED THESE, THE

1145
00:37:50,000 --> 00:37:53,280
CONTROL AND LGL1 WILD TYPE

1146
00:37:53,280 --> 00:37:54,080
EXPRESSING CELLS DIDN'T REALLY

1147
00:37:54,080 --> 00:37:55,720
LIKE IT, DIDN'T GROW WELL,

1148
00:37:55,720 --> 00:37:59,240
WHEREAS THE LGL2 CELLS CONTINUED

1149
00:37:59,240 --> 00:38:00,120
TO GROW WELL.

1150
00:38:00,120 --> 00:38:01,720
SO THIS GAVE US A CONNECTION

1151
00:38:01,720 --> 00:38:03,640
THAT MAYBE THERE'S A CONNECTION

1152
00:38:03,640 --> 00:38:08,880
BETWEEN NUTRIENT AND LGL2.

1153
00:38:08,880 --> 00:38:14,040
THIS IS SIMPLY REMOVING THE --

1154
00:38:14,040 --> 00:38:17,040
IF YOU DIAL -- YOU'LL HAVE --

1155
00:38:17,040 --> 00:38:18,840
THEN YOU GAIN THE PHENOTYPE.

1156
00:38:18,840 --> 00:38:20,440
AND WHAT YOU LOSE HERE ARE

1157
00:38:20,440 --> 00:38:30,480
THINGS LAKE AMINO ACI LIKE AMIN.

1158
00:38:30,480 --> 00:38:35,280
WE ALSO GENERATED LGL -- HE PUT

1159
00:38:35,280 --> 00:38:38,800
THEM IN XENOGRAPH TRANSPLANTS.

1160
00:38:38,800 --> 00:38:42,760
KNOCKDOWN OF LGL2 REALLY IMPAIRS

1161
00:38:42,760 --> 00:38:45,800
TUMOR -- IT IS A POSITIVE

1162
00:38:45,800 --> 00:38:49,040
REGULATOR OF GROWTH FOR THESE ER

1163
00:38:49,040 --> 00:38:50,240
POSITIVE BREAST CANCER CELLS.

1164
00:38:50,240 --> 00:38:52,600
SINCE THIS IS ALL NUTRITIONALLY

1165
00:38:52,600 --> 00:38:54,320
RELATED, WE ALSO VENTURED INTO

1166
00:38:54,320 --> 00:38:57,480
REALLY LOOKING AT THE

1167
00:38:57,480 --> 00:39:00,280
METABALOME, YOU SAID ANY -- WHEN

1168
00:39:00,280 --> 00:39:02,840
YOU ANALYZE ALL THE METABOLITES,

1169
00:39:02,840 --> 00:39:05,040
IT TURNS OUT THAT THE

1170
00:39:05,040 --> 00:39:09,040
INTRACELLULAR LEVELS WERE

1171
00:39:09,040 --> 00:39:10,320
DIFFERENTIALLY REGULATED BETWEEN

1172
00:39:10,320 --> 00:39:13,680
CONTROL AND KNOCK DOWN CELLS.

1173
00:39:13,680 --> 00:39:15,400
THE CELL CANNOT MAKE THEM SO

1174
00:39:15,400 --> 00:39:16,800
THEY RELY ON EXTRACELLULAR

1175
00:39:16,800 --> 00:39:20,120
SOURCE TO TAKE UP AMINO ACIDS

1176
00:39:20,120 --> 00:39:21,800
FOR BIOLOGICAL PROCESSES.

1177
00:39:21,800 --> 00:39:25,400
NOW AMONG THE NINE ESSENTIAL

1178
00:39:25,400 --> 00:39:33,440
AMINO ACIDS, SIGNIFICANT LOWER

1179
00:39:33,440 --> 00:39:34,760
IN KNOCK DOWN CELL THAN COMPARED

1180
00:39:34,760 --> 00:39:35,480
TO OTHERS.

1181
00:39:35,480 --> 00:39:36,560
SINCE THEY WERE MULTIPLE, YOU

1182
00:39:36,560 --> 00:39:38,920
ALSO WANTED TO KNOW IF ANY OF

1183
00:39:38,920 --> 00:39:40,920
THEM ARE SPECIFICALLY INVOLVED

1184
00:39:40,920 --> 00:39:42,160
OR A COLLECTION OF ALL OF THEM.

1185
00:39:42,160 --> 00:39:44,120
TO DO THAT HE LOWERED ALL THE

1186
00:39:44,120 --> 00:39:46,120
ESSENTIAL AMINO ACIDS AND THEN

1187
00:39:46,120 --> 00:39:47,960
ADDED BACK ONE AT A TIME TO SEE

1188
00:39:47,960 --> 00:39:50,840
WERE ANY OF THOSE RESCUE

1189
00:39:50,840 --> 00:39:53,680
PROLIFERATION.

1190
00:39:53,680 --> 00:39:56,600
THIS GAVE US A DIRECT INDICATION

1191
00:39:56,600 --> 00:39:57,840
COMPLETELY UNEXPECTED THAT

1192
00:39:57,840 --> 00:39:59,440
THERE'S SOME CONNECTION BETWEEN

1193
00:39:59,440 --> 00:40:01,600
A POLARITY PROTEIN LGL2 AND

1194
00:40:01,600 --> 00:40:03,400
LOSING UP TAKE BY THESE CELLS IN

1195
00:40:03,400 --> 00:40:04,280
THE WAY THEY USE IT.

1196
00:40:04,280 --> 00:40:05,800
SO THE QUESTION IS HOW DOES THIS

1197
00:40:05,800 --> 00:40:06,120
HAPPEN?

1198
00:40:06,120 --> 00:40:07,840
I'LL JUST SUMMARIZE IT SO I CAN

1199
00:40:07,840 --> 00:40:09,400
TELL YOU ABOUT SOME UNPUBLISHED

1200
00:40:09,400 --> 00:40:09,640
WORK.

1201
00:40:09,640 --> 00:40:11,600
WHAT HE FOUND WAS THAT THIS

1202
00:40:11,600 --> 00:40:13,400
POLARITY PROTEIN NOW FUNCTIONS

1203
00:40:13,400 --> 00:40:15,360
AS A TRAFFICKING REGULATOR.

1204
00:40:15,360 --> 00:40:18,480
IT BINDS TO AN AMINO ACID

1205
00:40:18,480 --> 00:40:20,760
TRANSPORTER ALSO KNOWN AS

1206
00:40:20,760 --> 00:40:22,360
LAT1 AND IT BINDS TO THE CYTOSOL

1207
00:40:22,360 --> 00:40:24,200
AND TRANSPORTS TO THE CELL

1208
00:40:24,200 --> 00:40:24,480
MEMBRANE.

1209
00:40:24,480 --> 00:40:29,720
IF YOU KNOCK DOWN LGL2, THE

1210
00:40:29,720 --> 00:40:30,200
LOCALIZATION CHANGES.

1211
00:40:30,200 --> 00:40:32,080
NOW IT ACCUMULATES IN THE

1212
00:40:32,080 --> 00:40:33,000
CYTOSOL, DOESN'T GET TO THE

1213
00:40:33,000 --> 00:40:33,280
MEMBRANE.

1214
00:40:33,280 --> 00:40:36,760
SO WHEN YOU HAVE HIGHER LEVELS

1215
00:40:36,760 --> 00:40:42,520
OF L TB. L2 -- AND THAT GIVES

1216
00:40:42,520 --> 00:40:43,880
THE CELL AN ADVANTAGE.

1217
00:40:43,880 --> 00:40:46,840
TO SUMMARIZE THESE, WHAT HE

1218
00:40:46,840 --> 00:40:47,720
FOUND -- IN THE MEMBRANE IT

1219
00:40:47,720 --> 00:40:51,360
BINDS TO A SNARE PROTEIN CALLED

1220
00:40:51,360 --> 00:40:52,880
YTK6 AND THAT FACILITATES IN

1221
00:40:52,880 --> 00:40:53,760
MEMBRANE INSERTION OF THESE

1222
00:40:53,760 --> 00:40:54,160
CELLS.

1223
00:40:54,160 --> 00:40:55,200
NOW THERE ARE A LOT OF QUESTIONS

1224
00:40:55,200 --> 00:40:56,640
HERE WE NEED TO UNDERSTAND

1225
00:40:56,640 --> 00:40:57,960
EXACTLY HOW THIS PROCESS WORKS,

1226
00:40:57,960 --> 00:41:00,640
AND THAT IS SOMETHING HE'S DOING

1227
00:41:00,640 --> 00:41:04,200
IN HIS OWN LAB IN JA JAPAN.

1228
00:41:04,200 --> 00:41:05,440
SO IN ADDITION TO REALLY

1229
00:41:05,440 --> 00:41:09,600
UNDERSTANDING THE ROLE FOR

1230
00:41:09,600 --> 00:41:12,440
LDR2 IN -- HE MADE AN

1231
00:41:12,440 --> 00:41:13,280
INTERESTING OBSERVATION, IT

1232
00:41:13,280 --> 00:41:14,280
TURNS OUT WHEN YOU HAVE HIGH

1233
00:41:14,280 --> 00:41:17,080
LEVELS OF LGL2, THESE DRUGS CAN

1234
00:41:17,080 --> 00:41:17,640
RESIST TAMOXIFEN.

1235
00:41:17,640 --> 00:41:19,800
SO THAT MEANT THAT MAYBE IT IS

1236
00:41:19,800 --> 00:41:22,120
INVOLVED IN SOME -- IN DRUG --

1237
00:41:22,120 --> 00:41:23,920
DEVELOPMENT OF DRUG RESISTANCE.

1238
00:41:23,920 --> 00:41:25,840
NOW FOR THOSE OF YOU THAT KNOW,

1239
00:41:25,840 --> 00:41:28,400
70% OF BREAST CANCERS ARE THE

1240
00:41:28,400 --> 00:41:30,160
ERPR POSITIVE SUBTYPE AND ABOUT

1241
00:41:30,160 --> 00:41:32,800
30% OF THEM WILL ACTUALLY

1242
00:41:32,800 --> 00:41:33,920
DEVELOP RESISTANCE TO TREATMENT

1243
00:41:33,920 --> 00:41:34,440
CURRENTLY USED.

1244
00:41:34,440 --> 00:41:37,040
IN THE CLINIC, THOSE 30% OF THE

1245
00:41:37,040 --> 00:41:40,920
PATIENTS RECEIVE CDK46 INHIBITOR

1246
00:41:40,920 --> 00:41:41,720
AS A TREATMENT PARADIGM FOR

1247
00:41:41,720 --> 00:41:42,600
THEM.

1248
00:41:42,600 --> 00:41:44,680
SO WE BEGAN ASKING IS THIS

1249
00:41:44,680 --> 00:41:48,480
REALLY RELEVANT IN THE --

1250
00:41:48,480 --> 00:41:49,240
CONTEXT AS WELL.

1251
00:41:49,240 --> 00:41:52,440
SO THESE ARE ALL ONGOING STUDIES

1252
00:41:52,440 --> 00:41:53,640
BEING DONE BY TALENTED POSTDOC

1253
00:41:53,640 --> 00:41:56,160
IN THE LAB, MIKE OLIPHANT, AND

1254
00:41:56,160 --> 00:41:57,680
I'M GOING TO SHARE WITH YOU AND

1255
00:41:57,680 --> 00:41:58,920
HAPPY TO TAKE ANY SUGGESTIONS ON

1256
00:41:58,920 --> 00:42:00,680
THIS PART OF THE PROJECT.

1257
00:42:00,680 --> 00:42:02,080
SO HE GENERATED A PANEL OF CELL

1258
00:42:02,080 --> 00:42:10,600
LINES THAT EXPRESS SLC7A5.

1259
00:42:10,600 --> 00:42:13,840
WHEN YOU OVEREXPRESS 7A5 THAT BY

1260
00:42:13,840 --> 00:42:15,040
ITSELF WAS SUFFICIENT TO GIVE A

1261
00:42:15,040 --> 00:42:18,920
THREE TO FIVE FOLD SHIFT IN

1262
00:42:18,920 --> 00:42:20,240
ISA50 VALUE, SO THE RED LINE

1263
00:42:20,240 --> 00:42:21,560
SHOWS THE OVEREXPRESSING CELLS

1264
00:42:21,560 --> 00:42:23,640
AND THE BLUE LINE IS THE CONTROL

1265
00:42:23,640 --> 00:42:25,880
CELLS, AND YOU CAN SEE THERE IS

1266
00:42:25,880 --> 00:42:28,480
APPROXIMATELY A FOUR PHOTO

1267
00:42:28,480 --> 00:42:31,000
INCREASE IN THE IC50 VALUES

1268
00:42:31,000 --> 00:42:34,080
MEANING THEY'RE -- THAN THE

1269
00:42:34,080 --> 00:42:34,360
REST.

1270
00:42:34,360 --> 00:42:36,600
THIS IS AN INDUCED RESISTANCE SO

1271
00:42:36,600 --> 00:42:39,600
THIS SUGGESTS 75 IS SUFFICIENT

1272
00:42:39,600 --> 00:42:41,160
TO CONFER RESISTANCE.

1273
00:42:41,160 --> 00:42:42,640
THEN THE QUESTION WAS IS IT

1274
00:42:42,640 --> 00:42:43,400
NATURALLY SELECTED.

1275
00:42:43,400 --> 00:42:46,560
SO WE WANTED TO LOOK AT SOME

1276
00:42:46,560 --> 00:42:49,040
SPECIFIC RESISTANT MODELS TO SEE

1277
00:42:49,040 --> 00:42:50,560
IS THERE -- SO WE COLLABORATED

1278
00:42:50,560 --> 00:42:54,200
WITH NICK TURNER TO GET THE CELL

1279
00:42:54,200 --> 00:42:56,640
LINES MADE WITH CDK46 AND WHEN

1280
00:42:56,640 --> 00:42:59,360
MIKE LOOKS AT THE SURFACE

1281
00:42:59,360 --> 00:43:01,440
LEVELS, THEY'RE SIGNIFICANTLY

1282
00:43:01,440 --> 00:43:03,200
HIGHER IN THESE MODELS COMPARED

1283
00:43:03,200 --> 00:43:04,840
TO THE SENSITIVE ONES,

1284
00:43:04,840 --> 00:43:06,160
SUGGESTING THE FACT THERE IS

1285
00:43:06,160 --> 00:43:07,040
PROBABLY SELECTION FOR IT DURING

1286
00:43:07,040 --> 00:43:08,920
THE DEVELOPMENT OF RESISTANCE IN

1287
00:43:08,920 --> 00:43:09,440
THESE CELL LINES.

1288
00:43:09,440 --> 00:43:12,200
THE MORE POWERFUL DATA CAME WHEN

1289
00:43:12,200 --> 00:43:14,880
BEGAN A COLLABORATION WITH NICK

1290
00:43:14,880 --> 00:43:18,160
WAGLEY'S GRAWP AT DANA FARBER.

1291
00:43:18,160 --> 00:43:21,320
NICK HAS BEEN COLLABORATING

1292
00:43:21,320 --> 00:43:25,880
SAMPLES AND ANALYZING THE --

1293
00:43:25,880 --> 00:43:31,880
WITH THE DEVELOPMENT OF C

1294
00:43:31,880 --> 00:43:32,720
CD46 INHIBITORS IN THE CLINIC.

1295
00:43:32,720 --> 00:43:34,040
IN THE FIVE PAIRS WE LOOKED AT,

1296
00:43:34,040 --> 00:43:36,360
IN ALL CASES, THERE WAS A

1297
00:43:36,360 --> 00:43:37,960
SIGNIFICANT INCREASE, A

1298
00:43:37,960 --> 00:43:38,560
NOTICEABLE INCREASE, I SHOULD

1299
00:43:38,560 --> 00:43:39,560
SAY, IT'S TOO SMALL A NUMBER TO

1300
00:43:39,560 --> 00:43:43,040
CALL IT SIGNIFICANT, INCREASE IN

1301
00:43:43,040 --> 00:43:45,800
THE LEVELS OF 7A5 PRE AND POST

1302
00:43:45,800 --> 00:43:47,720
TREATMENT.

1303
00:43:47,720 --> 00:43:49,400
ALTHOUGH THIS LOOKS LIKE FIVE

1304
00:43:49,400 --> 00:43:50,960
SAMPLES, THESE ARE VERY PRECIOUS

1305
00:43:50,960 --> 00:43:51,880
SAMPLES TO COME BY.

1306
00:43:51,880 --> 00:43:53,480
SO THIS IS REALLY REASSURING FOR

1307
00:43:53,480 --> 00:43:56,040
US TO KNOW THAT 7A5 MAY ACTUALLY

1308
00:43:56,040 --> 00:43:57,880
BE DRIVING RESISTANCE IN THESE

1309
00:43:57,880 --> 00:43:58,480
PATIENTS.

1310
00:43:58,480 --> 00:43:59,880
IT TURNS OUT IF YOU BREAK THEM

1311
00:43:59,880 --> 00:44:02,920
DOWN INTO PATIENTS WHO HAVE A

1312
00:44:02,920 --> 00:44:03,880
QUIET RESISTANCE THAN HAVE

1313
00:44:03,880 --> 00:44:07,600
PATIENT WHO HAVE -- THE INTR,

1314
00:44:07,600 --> 00:44:09,040
THE -- COMPARED TO THE SENSITIVE

1315
00:44:09,040 --> 00:44:09,520
PATIENTS.

1316
00:44:09,520 --> 00:44:10,600
SO ARMED BY THIS INFORMATION

1317
00:44:10,600 --> 00:44:12,000
THAT THIS IS PROBABLY A DRIVER

1318
00:44:12,000 --> 00:44:15,200
OF RESISTANCE, MIKE HAS BEGUN A

1319
00:44:15,200 --> 00:44:16,840
SERIES OF STUDIES TO REALLY GO

1320
00:44:16,840 --> 00:44:18,520
DOWN THE PATH OF UNDERSTANDING

1321
00:44:18,520 --> 00:44:19,400
THE MECHANISM BY WHICH THIS MAY

1322
00:44:19,400 --> 00:44:19,960
HAPPEN.

1323
00:44:19,960 --> 00:44:21,880
SO LEUCINE, AS SOME OF YOU MAY

1324
00:44:21,880 --> 00:44:23,800
KNOW, IS A BRANCH AMINO ACID SO

1325
00:44:23,800 --> 00:44:25,680
THERE ARE SOME SIMPLE KITS YOU

1326
00:44:25,680 --> 00:44:28,120
CAN USE TO MEASURE BRANCH AMINO

1327
00:44:28,120 --> 00:44:29,560
ACID LEVELS IN CELLS.

1328
00:44:29,560 --> 00:44:32,120
SO MIKE TOOK HIS OVEREXPRESSING

1329
00:44:32,120 --> 00:44:36,040
CELL LINES AND SPONTANEOUS

1330
00:44:36,040 --> 00:44:38,840
RESISTANT MODEL, IN BOTH CASES,

1331
00:44:38,840 --> 00:44:40,080
THE LEVELS GO UP.

1332
00:44:40,080 --> 00:44:42,240
NOT ONLY THE LEVELS TRANSPORTED

1333
00:44:42,240 --> 00:44:43,680
HIGHER IN THESE CELLS, THEY

1334
00:44:43,680 --> 00:44:45,000
FUNCTIONALLY ARE WORKING MORE BY

1335
00:44:45,000 --> 00:44:47,040
TAKING UP MORE AMINO ACIDS

1336
00:44:47,040 --> 00:44:48,000
INSIDE THE CELL.

1337
00:44:48,000 --> 00:44:50,280
IS THE INCREASING LEVEL OF AMINO

1338
00:44:50,280 --> 00:44:51,800
ACID, IS THAT WHAT IS DRIVING

1339
00:44:51,800 --> 00:44:52,120
THE PHENOTYPE?

1340
00:44:52,120 --> 00:44:55,400
TO TEST THAT, WHAT MIKE DID IS A

1341
00:44:55,400 --> 00:44:56,400
SIMPLE EXPERIMENT.

1342
00:44:56,400 --> 00:44:57,840
YOU COMPARE COMPLETE MEDIA

1343
00:44:57,840 --> 00:44:59,160
WHETHER IT'S NORMAL LEVELS OF

1344
00:44:59,160 --> 00:45:00,600
LEUCINE, TO THE DRUG RESPONSE IN

1345
00:45:00,600 --> 00:45:02,440
A MEDIA THAT HAS ONE-TENTH THE

1346
00:45:02,440 --> 00:45:03,360
AMOUNT OF LEUCINE.

1347
00:45:03,360 --> 00:45:04,480
SO IF YOU DROP THE LEUCINE

1348
00:45:04,480 --> 00:45:06,040
LEVELS THEN THE RESISTANCE

1349
00:45:06,040 --> 00:45:07,800
DROPS, THE CELLS BECOME A LITTLE

1350
00:45:07,800 --> 00:45:09,680
BIT MORE SENSITIVE, GIVING US

1351
00:45:09,680 --> 00:45:11,680
THE INDICATION THAT IN FACT

1352
00:45:11,680 --> 00:45:13,080
TOGGLING THE LEUCINE LEVELS

1353
00:45:13,080 --> 00:45:15,960
INSIDE THE CELL CAN TOGGLE IN

1354
00:45:15,960 --> 00:45:17,840
WHICH THE WAY THE CELLS CAN

1355
00:45:17,840 --> 00:45:18,080
FIGHT.

1356
00:45:18,080 --> 00:45:20,600
THOSE OF YOU, LEUCINE WHEN IT

1357
00:45:20,600 --> 00:45:22,720
GET INTO THE CELL IS AN

1358
00:45:22,720 --> 00:45:25,600
IMPORTANT INDICATOR OR MTOR

1359
00:45:25,600 --> 00:45:25,880
SIGNALING.

1360
00:45:25,880 --> 00:45:27,200
SO WE THOUGHT MAYBE THIS BIOLOGY

1361
00:45:27,200 --> 00:45:29,160
IS GOING TO MTOR.

1362
00:45:29,160 --> 00:45:31,640
IT'S TRUE WHEN YOU ADD LEUCINE,

1363
00:45:31,640 --> 00:45:33,520
WE ASKED IF YOU TREAT THESE

1364
00:45:33,520 --> 00:45:36,440
CELLS WITH INHIBITOR DO YOU SEE

1365
00:45:36,440 --> 00:45:38,600
A SENSITIVITY BETWEEN

1366
00:45:38,600 --> 00:45:40,280
OVEREXPRESSING CELLS,

1367
00:45:40,280 --> 00:45:42,400
DIFFERENTIAL BETWEEN -- AND

1368
00:45:42,400 --> 00:45:43,920
THE -- SO THEY ARE EQUALLY

1369
00:45:43,920 --> 00:45:44,400
SENSITIVE.

1370
00:45:44,400 --> 00:45:46,400
SO THIS TELLS US THAT THE

1371
00:45:46,400 --> 00:45:47,240
INCREASED CONCENTRATION OF

1372
00:45:47,240 --> 00:45:49,440
LEUCINE IN THESE RESISTANT CELLS

1373
00:45:49,440 --> 00:45:52,160
CONFERRED BY 7A5 OVEREXPRESSION

1374
00:45:52,160 --> 00:45:54,080
IS PROBABLY GOING THROUGH

1375
00:45:54,080 --> 00:45:56,080
SOMETHING ELSE OTHER THAN MTOR

1376
00:45:56,080 --> 00:45:56,320
BIOLOGY.

1377
00:45:56,320 --> 00:45:57,840
SO THE QUESTION IS WHAT ARE

1378
00:45:57,840 --> 00:45:58,040
THEY.

1379
00:45:58,040 --> 00:46:00,080
I THINK THIS IS AN EMERGING --

1380
00:46:00,080 --> 00:46:02,320
IT'S A PROJECT THAT IS ONGOING,

1381
00:46:02,320 --> 00:46:05,480
AND SO MIKE TOOK A TOTAL

1382
00:46:05,480 --> 00:46:07,400
METABALOME ANALYSIS, LOOKED AT

1383
00:46:07,400 --> 00:46:08,760
ALL THE ME TAB LOAN IN THESE

1384
00:46:08,760 --> 00:46:10,640
CELLS, AND SURPRISINGLY WHEN HE

1385
00:46:10,640 --> 00:46:13,040
LOOKED AT IT THE THE IPT

1386
00:46:13,040 --> 00:46:14,440
IMMEDIATES WERE ACTUALLY

1387
00:46:14,440 --> 00:46:20,760
UPREGULATED IN THESE CELLS, ALL

1388
00:46:20,760 --> 00:46:21,440
UP IN THESE CELLS.

1389
00:46:21,440 --> 00:46:23,680
BY CONTRAST, NONE OF THE TCA

1390
00:46:23,680 --> 00:46:25,000
CYCLE INTERMEDIATES WERE

1391
00:46:25,000 --> 00:46:26,000
ACTUALLY CHANGING IN THESE

1392
00:46:26,000 --> 00:46:26,360
CELLS.

1393
00:46:26,360 --> 00:46:28,120
SO THIS TOLD US THERE IS SOME

1394
00:46:28,120 --> 00:46:31,400
CONNECTION BETWEEN GLYCOLYSIS

1395
00:46:31,400 --> 00:46:34,200
AND THE AMINO ACID TRANSFORMER.

1396
00:46:34,200 --> 00:46:35,480
WE DON'T KNOW EXACTLY HOW IT

1397
00:46:35,480 --> 00:46:36,480
WORKS BUT WE'RE BEGINNING TO

1398
00:46:36,480 --> 00:46:36,920
UNDERSTAND IT.

1399
00:46:36,920 --> 00:46:39,000
SO THIS IS ALL IN CELL CULTURE.

1400
00:46:39,000 --> 00:46:42,200
SO SINCE WE HAVE HUGE AMOUNTS,

1401
00:46:42,200 --> 00:46:44,080
THIS IS DRIVEN BY CULTURE

1402
00:46:44,080 --> 00:46:45,680
CONDITIONS, SO WHAT MIKE DID WAS

1403
00:46:45,680 --> 00:46:52,000
HE USED THE SAME CCF7, GENERATED

1404
00:46:52,000 --> 00:46:53,720
XENOGRAPH TUMORS IN THE MAMMARY

1405
00:46:53,720 --> 00:46:55,680
FAT PAD AND THEN ANALYZED THE

1406
00:46:55,680 --> 00:46:56,920
METABALOME OF THOSE CELLS, AND

1407
00:46:56,920 --> 00:46:58,400
WHAT HE FOUND IS EXACTLY THE

1408
00:46:58,400 --> 00:47:01,360
SAME THING.

1409
00:47:01,360 --> 00:47:07,040
 -- GO UP BUT NOT THE TCS CYCLE.

1410
00:47:07,040 --> 00:47:09,360
SO -- OR TAKING UP MORE GLUCOSE,

1411
00:47:09,360 --> 00:47:10,800
AND WE DON'T KNOW WHAT THAT IS.

1412
00:47:10,800 --> 00:47:12,480
TO TEST IT, MIKE DID A SERIES OF

1413
00:47:12,480 --> 00:47:12,880
EXPERIMENTS.

1414
00:47:12,880 --> 00:47:14,560
HE MEASURED THE INTRACELLULAR

1415
00:47:14,560 --> 00:47:16,160
LEVELS OF GLUCOSE BY USING A

1416
00:47:16,160 --> 00:47:18,800
LABELED GLUCOSE, IN THIS CASE

1417
00:47:18,800 --> 00:47:20,440
2DG, AND ASKED WHETHER THE CELLS

1418
00:47:20,440 --> 00:47:21,760
HAVE A DEFINITE ABILITY TO TAKE

1419
00:47:21,760 --> 00:47:22,880
UP MORE GLUCOSE AND THE ANSWER

1420
00:47:22,880 --> 00:47:23,960
IS YES.

1421
00:47:23,960 --> 00:47:25,640
SO BOTH THE OVEREXPRESSING CELLS

1422
00:47:25,640 --> 00:47:28,120
AND THE SPONTANEOUS RESISTANT

1423
00:47:28,120 --> 00:47:30,120
MODEL CAN TAKE UP MORE GLUCOSE.

1424
00:47:30,120 --> 00:47:33,440
WE ALSO KNOW THE

1425
00:47:33,440 --> 00:47:35,080
GLUT1 TRANSPORTERS ALSO GO UP SO

1426
00:47:35,080 --> 00:47:37,800
THERE IS A BIOLOGY HERE

1427
00:47:37,800 --> 00:47:40,760
EMERGING, IT NOW CONNECTED TO

1428
00:47:40,760 --> 00:47:41,240
GLYCOLYSIS.

1429
00:47:41,240 --> 00:47:43,880
SO THE WAY THE CELL USES THESE

1430
00:47:43,880 --> 00:47:44,880
INTERMEDIATES IS REALLY TO

1431
00:47:44,880 --> 00:47:46,640
CHANGE THE BIOENERGETICS OF

1432
00:47:46,640 --> 00:47:48,320
THESE CELLS, SO WE BEGAN TESTING

1433
00:47:48,320 --> 00:47:51,760
TO SEE IS THERE A REAL

1434
00:47:51,760 --> 00:47:53,840
DIFFERENCE BETWEEN THE CONTROL

1435
00:47:53,840 --> 00:47:54,400
AND OVEREXPRESSING CELLS.

1436
00:47:54,400 --> 00:47:56,480
WE PLEASURED USING SEAHORSE

1437
00:47:56,480 --> 00:47:56,920
ANALYSIS.

1438
00:47:56,920 --> 00:48:00,120
BOTH OCR, OXYGEN CONSUMPTION

1439
00:48:00,120 --> 00:48:02,320
RATE -- ARE INCREASED IN THESE

1440
00:48:02,320 --> 00:48:02,520
CELLS.

1441
00:48:02,520 --> 00:48:04,760
I CAN ANSWER QUESTIONS ABOUT WHY

1442
00:48:04,760 --> 00:48:07,160
THIS IS THE CASE BECAUSE YOU

1443
00:48:07,160 --> 00:48:10,400
WOULD EXPECT ECAR LEVELS TO GO

1444
00:48:10,400 --> 00:48:13,840
UP WITH GLYCOLYSIS BUT NOT

1445
00:48:13,840 --> 00:48:14,880
CONTROL LEVELS.

1446
00:48:14,880 --> 00:48:16,640
YOU CAN MAP THESE TWO VALUES AND

1447
00:48:16,640 --> 00:48:17,840
CREATE AN ENERGY MAP AND WHAT WE

1448
00:48:17,840 --> 00:48:19,840
NOTE IS WHEN WE CREATE AN ENERGY

1449
00:48:19,840 --> 00:48:22,440
MAP, THE 7FA WORKS

1450
00:48:22,440 --> 00:48:24,160
OVEREXPRESSING CELLS EXIST IN A

1451
00:48:24,160 --> 00:48:26,960
HIGH ENERGETIC STATE, WHEREAS

1452
00:48:26,960 --> 00:48:31,680
THE CONTROL WERE IN A LOWER AND

1453
00:48:31,680 --> 00:48:32,720
ENERGETIC -- AND THE WAY THEY

1454
00:48:32,720 --> 00:48:33,800
CAN RESIST TREATMENT.

1455
00:48:33,800 --> 00:48:36,320
SO WHAT MIKE DID WAS TO REALLY

1456
00:48:36,320 --> 00:48:39,480
TAKE THESE SOLES, AND THEN

1457
00:48:39,480 --> 00:48:40,920
MEASURE OXYGEN CONSUMPTION RATE

1458
00:48:40,920 --> 00:48:42,600
TO SEE DOES THE TREATMENT DO

1459
00:48:42,600 --> 00:48:44,440
ANYTHING TO THESE CELLS.

1460
00:48:44,440 --> 00:48:47,160
IT TURNS OUT THE RESULTS WERE

1461
00:48:47,160 --> 00:48:48,200
COUNTERINTUITIVE AND SURPRISING.

1462
00:48:48,200 --> 00:48:49,880
THEY ACTUALLY GO TO EVEN HIGHER

1463
00:48:49,880 --> 00:48:52,560
ENERGY STATE, AS IF THEY ARE

1464
00:48:52,560 --> 00:48:56,400
REVVING UP THEIR SYSTEM TO RUN

1465
00:48:56,400 --> 00:48:56,840
AWAY.

1466
00:48:56,840 --> 00:48:59,280
SO THIS REALLY PUTS US IN A VERY

1467
00:48:59,280 --> 00:49:00,280
CLEAR PATH.

1468
00:49:00,280 --> 00:49:01,640
MAYBE THE ENERGY STATE OF THE

1469
00:49:01,640 --> 00:49:05,960
CELL IS REALLY -- IF THAT'S THE

1470
00:49:05,960 --> 00:49:08,960
CASE, DO YOU NOW RESTORE THE

1471
00:49:08,960 --> 00:49:09,200
BIOLOGY.

1472
00:49:09,200 --> 00:49:11,320
TO DO THAT YOU CAN INHIBIT

1473
00:49:11,320 --> 00:49:15,120
GLUCOSE UPDATE BY USING A 2DG

1474
00:49:15,120 --> 00:49:16,760
GLUCOSE TO BLOCK GLYCOLYSIS.

1475
00:49:16,760 --> 00:49:19,200
WHEN YOU TREAT THESE CELLS WITH

1476
00:49:19,200 --> 00:49:20,800
2GD YOU SHUT DOWN THE ECAR AND

1477
00:49:20,800 --> 00:49:23,840
THE OCR BIG TIME AND YOU CREATE

1478
00:49:23,840 --> 00:49:25,400
AN ENERGY MAP.

1479
00:49:25,400 --> 00:49:28,840
UNDER THESE CONDITIONS,

1480
00:49:28,840 --> 00:49:29,560
RESISTANCE IS SIGNIFICANTLY

1481
00:49:29,560 --> 00:49:29,960
IMPAIRED.

1482
00:49:29,960 --> 00:49:31,160
SO WE KNOW WE CAN TOGGLE THESE

1483
00:49:31,160 --> 00:49:31,480
THINGS.

1484
00:49:31,480 --> 00:49:32,840
NOW THE BIG CONNECTION FOR US WE

1485
00:49:32,840 --> 00:49:36,600
NEED TO UNDERSTAND IS, HOW DOES

1486
00:49:36,600 --> 00:49:38,160
AP AMINO ACID TRANSPORTER,

1487
00:49:38,160 --> 00:49:41,000
CONNECT AND REINQUIRE THE

1488
00:49:41,000 --> 00:49:42,360
GLYCOLYSIS IN THOSE THEES CELLS

1489
00:49:42,360 --> 00:49:44,760
AND WE ARE NOW ACTIVELY WORKING

1490
00:49:44,760 --> 00:49:45,560
TO UNDERSTAND THIS BIOLOGY.

1491
00:49:45,560 --> 00:49:48,880
SO THESE STUDIES WITH LGL AND

1492
00:49:48,880 --> 00:49:50,320
7A5 HAS REALLY PUT NUSS A PATH

1493
00:49:50,320 --> 00:49:52,120
WITH HOW WE CAN STUDY POLARITY

1494
00:49:52,120 --> 00:49:57,480
PROTEINS IN THE CONTEXT OF

1495
00:49:57,480 --> 00:49:59,680
CELL -- AND POTENTIALLY EVEN THE

1496
00:49:59,680 --> 00:50:03,240
CELL SURFACE ONLY, AND ALL THESE

1497
00:50:03,240 --> 00:50:05,080
COMBINED CAN HAVE AN IMPACT IN

1498
00:50:05,080 --> 00:50:06,080
DRUG RESISTANCE, METASTASES AND

1499
00:50:06,080 --> 00:50:07,680
THE WAY THE CELL SEES THE

1500
00:50:07,680 --> 00:50:08,080
MICROENVIRONMENT.

1501
00:50:08,080 --> 00:50:08,920
THESE ARE SOME OF THE STUDIES

1502
00:50:08,920 --> 00:50:10,440
AND PROJECTS THAT WE ARE

1503
00:50:10,440 --> 00:50:11,520
CURRENTLY INITIATING IN THE LAB

1504
00:50:11,520 --> 00:50:13,360
HERE AT LCBG.

1505
00:50:13,360 --> 00:50:19,440
SO IN SUMMARY, AS I MENTIONED,

1506
00:50:19,440 --> 00:50:20,800
THE RESEARCH PROGRAM IN OUR LAB

1507
00:50:20,800 --> 00:50:22,160
REALLY IS AN INTEGRATION BETWEEN

1508
00:50:22,160 --> 00:50:23,280
BASIC AND TRANSLATION RESEARCH,

1509
00:50:23,280 --> 00:50:24,400
WE GO BACK AND FORTH.

1510
00:50:24,400 --> 00:50:26,560
AND THE CELL POLARITY STUDIES

1511
00:50:26,560 --> 00:50:31,400
REALLY UNDERSTANDING SIGN SIGNAG

1512
00:50:31,400 --> 00:50:34,840
SUR FA SOME IN ORGANOIDS, USING

1513
00:50:34,840 --> 00:50:36,880
THEM FOR EITHER MODELING

1514
00:50:36,880 --> 00:50:39,080
CLINICAL RESPONSE OR

1515
00:50:39,080 --> 00:50:43,040
UNDERSTANDING -- FINDING THERAPY

1516
00:50:43,040 --> 00:50:43,560
RESISTANCE IN THE CLINIC.

1517
00:50:43,560 --> 00:50:45,400
SO ALL OF THIS CANNOT BE DONE

1518
00:50:45,400 --> 00:50:47,880
WITHOUT POWERFUL COLLABORATIONS

1519
00:50:47,880 --> 00:50:48,760
AND THE RESEARCH GREUN.

1520
00:50:48,760 --> 00:50:50,120
SO THESE ARE THE COLLABORATORS

1521
00:50:50,120 --> 00:50:51,200
WHO HELPED US IN DIFFERENT PARTS

1522
00:50:51,200 --> 00:50:52,120
OF THE PROJECT IN THE PAST.

1523
00:50:52,120 --> 00:50:54,240
SO ALL THE ORGANOID WORK THAT

1524
00:50:54,240 --> 00:50:56,560
I'VE PRESENTED TODAY WAS DONE

1525
00:50:56,560 --> 00:50:58,520
BEFORE I GOT HERE.

1526
00:50:58,520 --> 00:51:04,400
IT'S FROM BIDMC HEAD AND NECK

1527
00:51:04,400 --> 00:51:10,440
AND ONCOLOGY WITH HELP FROM

1528
00:51:10,440 --> 00:51:11,920
MANUEL HIDALGO.

1529
00:51:11,920 --> 00:51:18,720
CELL PRO LA POLARITY AND DRUG

1530
00:51:18,720 --> 00:51:23,760
RESISTANCE WITH MARCIA HAGIS,

1531
00:51:23,760 --> 00:51:26,840
MYLES BROWN AND FORREST WHITE.

1532
00:51:26,840 --> 00:51:27,920
THE MEMBERS.

1533
00:51:27,920 --> 00:51:28,960
NEW TEAM ARE LISTED THERE, AND

1534
00:51:28,960 --> 00:51:30,320
MIKE IS STILL IN BOSTON FOR

1535
00:51:30,320 --> 00:51:31,800
FAMILY REASONS AND CONTINUING

1536
00:51:31,800 --> 00:51:33,360
THE PROJECT, AND WE HAVE NOW

1537
00:51:33,360 --> 00:51:37,920
INITIATED A STRONG COLLABORATION

1538
00:51:37,920 --> 00:51:40,240
HERE AND GETTING SAMPLES TO

1539
00:51:40,240 --> 00:51:41,440
GENERATE ORGANOID MODELS BOTH

1540
00:51:41,440 --> 00:51:43,640
FOR THE BIOBANK APPROACH AND FOR

1541
00:51:43,640 --> 00:51:45,320
THE TCR DISCOVERY STUDIES.

1542
00:51:45,320 --> 00:51:47,680
AND THE MEMBERS OF THE WORK I

1543
00:51:47,680 --> 00:51:48,760
PRESENTED, I MENTIONED THEM

1544
00:51:48,760 --> 00:51:49,160
ALONG THE WAY.

1545
00:51:49,160 --> 00:51:52,040
AND I CAN STOP HERE, AND BEFORE

1546
00:51:52,040 --> 00:51:53,280
I TAKE ANY QUESTIONS I WANT TO

1547
00:51:53,280 --> 00:51:54,720
THANK ALL OF YOU FOR BEING HERE

1548
00:51:54,720 --> 00:51:56,320
BECAUSE IT IS AMAZING TO TALK TO

1549
00:51:56,320 --> 00:51:58,320
LIVE AUDIENCE FACE-TO-FACE THAN

1550
00:51:58,320 --> 00:51:58,960
SEEING A MONITOR.

1551
00:51:58,960 --> 00:52:00,040
SO THANK YOU FOR MAKING THE

1552
00:52:00,040 --> 00:52:00,880
EFFORT OF BEING HERE.

1553
00:52:00,880 --> 00:52:10,960
[APPLAUSE]

1554
00:52:10,960 --> 00:52:13,920
>>WE'D LIKE TO OPEN IT UP
TO

1555
00:52:13,920 --> 00:52:15,200
QUESTIONS AT THIS PARTICULAR

1556
00:52:15,200 --> 00:52:15,760
POINT IN TIME.

1557
00:52:15,760 --> 00:52:16,600
WHILE YOU'RE GATHERING YOUR

1558
00:52:16,600 --> 00:52:18,240
THOUGHTS ON THAT, YOU TALKED

1559
00:52:18,240 --> 00:52:18,960
ABOUT -- JUST ABOUT EVERYTHING

1560
00:52:18,960 --> 00:52:22,480
THAT YOU COULD DO WITH ORGANO

1561
00:52:22,480 --> 00:52:23,160
ORGANOIDS, BUT YOU DIDN'T

1562
00:52:23,160 --> 00:52:25,160
MENTION ANYTHING ABOUT

1563
00:52:25,160 --> 00:52:26,120
EPIGENETICS.

1564
00:52:26,120 --> 00:52:28,520
SO ARE YOU DOING ANYTHING OR

1565
00:52:28,520 --> 00:52:29,640
THINKING ABOUT WORKING IN THAT

1566
00:52:29,640 --> 00:52:30,720
AREA AT ALL?

1567
00:52:30,720 --> 00:52:34,160
>>NO, BECAUSE WE'RE NOT
GOOD AT

1568
00:52:34,160 --> 00:52:34,680
IT.

1569
00:52:34,680 --> 00:52:35,920
SO BUT WE ARE HAPPY TO HELP

1570
00:52:35,920 --> 00:52:37,760
ANYBODY WHO WANTS TO DO.

1571
00:52:37,760 --> 00:52:42,760
>>OKAY.

1572
00:52:42,760 --> 00:52:45,040
>>HI.

1573
00:52:45,040 --> 00:52:47,240
THIS IS -- FROM SURGERY BRANCH.

1574
00:52:47,240 --> 00:52:51,160
THE PAST FEW YEARS WE'VE BEEN

1575
00:52:51,160 --> 00:52:52,360
GROWING FROM OUR RESECTIONS AND

1576
00:52:52,360 --> 00:52:54,000
WE'VE BEEN TESTING THEM ALONG

1577
00:52:54,000 --> 00:52:55,320
OUR NEOANTIGEN PIPELINE, RIGHT,

1578
00:52:55,320 --> 00:52:58,440
TO FIND THE REACTIVITIES.

1579
00:52:58,440 --> 00:52:59,840
THE CHALLENGE WE'RE CURRENTLY

1580
00:52:59,840 --> 00:53:02,600
FACING IS FINDING MANY

1581
00:53:02,600 --> 00:53:03,640
REACTIVITIES AGAINST ORGANOID

1582
00:53:03,640 --> 00:53:05,720
THAT ARE NOT NEW ANTIGENS, SO

1583
00:53:05,720 --> 00:53:07,760
THE TARGETS ARE UNKNOWN.

1584
00:53:07,760 --> 00:53:13,040
HAVE YOU WORKED ON WHAT --

1585
00:53:13,040 --> 00:53:14,000
IDENTIFYING THOSE TUMOR TARGETS

1586
00:53:14,000 --> 00:53:15,640
THAT THESE TCRs ARE SEEING?

1587
00:53:15,640 --> 00:53:16,960
>>THAT'S A MILLION DOLLAR

1588
00:53:16,960 --> 00:53:20,560
QUESTION BECAUSE IT IS SUCH A

1589
00:53:20,560 --> 00:53:22,120
DIFFICULT TASK, BUT WE HAVE

1590
00:53:22,120 --> 00:53:24,760
INITIATED A COLLABORATION USING

1591
00:53:24,760 --> 00:53:29,440
ONE OF THE TCRs, SO HE HAS A

1592
00:53:29,440 --> 00:53:30,960
DISCOVERY PIPELINE THAT HAS THE

1593
00:53:30,960 --> 00:53:34,000
ENTIRE CODED GENOME PRESENTED ON

1594
00:53:34,000 --> 00:53:36,520
THE SURFACE AS SHORT PEPTIDES.

1595
00:53:36,520 --> 00:53:40,160
IN A CELL-BASED ASSAY, YOU CAN

1596
00:53:40,160 --> 00:53:41,960
ENGINEER THESE TCR IN A T WELL

1597
00:53:41,960 --> 00:53:43,440
AND USE IT AS BAIT TO FIND OUT

1598
00:53:43,440 --> 00:53:44,520
WHAT IT BINDS TO, KEEP IN MIND

1599
00:53:44,520 --> 00:53:45,800
THESE ARE JUST PEPTIDES

1600
00:53:45,800 --> 00:53:46,040
PRESENTED.

1601
00:53:46,040 --> 00:53:47,560
IF THE PEPTIDE IS MODIFIED IN

1602
00:53:47,560 --> 00:53:49,000
ANY FORM OR SHAPE, THAT IS NOT

1603
00:53:49,000 --> 00:53:51,120
REPRESENTED HERE.

1604
00:53:51,120 --> 00:53:52,040
RIGHT?

1605
00:53:52,040 --> 00:53:54,000
SO WE CAN ALSO TAKE THE TCRs

1606
00:53:54,000 --> 00:53:56,280
AND DO COMPUTATIONALLY ASK WHAT

1607
00:53:56,280 --> 00:53:57,480
MAY BE THE LIKELY TARGET.

1608
00:53:57,480 --> 00:53:59,720
SO WE DID BOTH.

1609
00:53:59,720 --> 00:54:03,360
SO FOR ONE OF THE TCRs.

1610
00:54:03,360 --> 00:54:05,040
THE COMPUTATIONAL APPROACH GAVE

1611
00:54:05,040 --> 00:54:06,840
A PEPTIDE FOR AN ANTIGEN THAT

1612
00:54:06,840 --> 00:54:10,120
WAS A COMMON ONCOGENE, TO SEE IF

1613
00:54:10,120 --> 00:54:12,800
IT WOULD STIMULATE AND IT HAD NO

1614
00:54:12,800 --> 00:54:16,240
EFFECT.

1615
00:54:16,240 --> 00:54:17,480
THEN THROUGH STEVE'S GROUP HE

1616
00:54:17,480 --> 00:54:19,000
FOUND A PEPTIDE THAT IS

1617
00:54:19,000 --> 00:54:20,200
TRANSCRIBED IN THE REVERSE

1618
00:54:20,200 --> 00:54:23,800
DIRECTION OF A CODING GENE.

1619
00:54:23,800 --> 00:54:26,240
SO IT IS -- IT'S COMPLETELY

1620
00:54:26,240 --> 00:54:26,880
COUNTERINTUITIVE WHAT YOU WOULD

1621
00:54:26,880 --> 00:54:28,000
THINK ABOUT.

1622
00:54:28,000 --> 00:54:29,440
THIS PEPTIDE YOU WOULDN'T EVEN

1623
00:54:29,440 --> 00:54:30,760
THINK THAT WOULD EXIST BECAUSE

1624
00:54:30,760 --> 00:54:32,000
IT'S A SHORT PEPTIDE IN THE

1625
00:54:32,000 --> 00:54:32,760
REVERSE DIRECTION OF A GENE.

1626
00:54:32,760 --> 00:54:34,240
SO WHETHER THAT IS THE GENE OR

1627
00:54:34,240 --> 00:54:35,600
NOT, THE PEPTIDE OR NOT, WE

1628
00:54:35,600 --> 00:54:37,720
DON'T KNOW, BUT THAT WAS THE

1629
00:54:37,720 --> 00:54:38,600
STRONGEST HIT AND THE ONLY HIT

1630
00:54:38,600 --> 00:54:39,200
HE COULD FIND.

1631
00:54:39,200 --> 00:54:40,480
SO IT ONLY TELLS US THAT YOU

1632
00:54:40,480 --> 00:54:42,520
HAVE TO TAKE COMPLETELY UNBIASED

1633
00:54:42,520 --> 00:54:45,040
APPROACH TO GO AFTER THEM AND

1634
00:54:45,040 --> 00:54:46,360
EACH TCR WOULD BE A TASK ON ITS

1635
00:54:46,360 --> 00:54:46,680
OWN.

1636
00:54:46,680 --> 00:54:47,840
SO I DON'T HAVE AN ANSWER BUT I

1637
00:54:47,840 --> 00:54:48,880
THINK IT IS FEASIBLE.

1638
00:54:48,880 --> 00:54:51,040
>>ALL RIGHT.

1639
00:54:51,040 --> 00:54:52,600
THE OTHER THING WE'VE BEEN DOING

1640
00:54:52,600 --> 00:54:54,760
IS WHATEVER THE HARVEST SITE IS,

1641
00:54:54,760 --> 00:54:58,960
SAY IF WE TAKE COLORECTAL

1642
00:54:58,960 --> 00:55:00,080
METASTATIC TO THE LUNG, WE TRY

1643
00:55:00,080 --> 00:55:01,760
TO GROW THE NORMAL LUNG AS AN

1644
00:55:01,760 --> 00:55:03,320
ORGANOID AND USE AS A CONTROL.

1645
00:55:03,320 --> 00:55:05,160
DO YOU HAVE ANY EXPERIENCE IN

1646
00:55:05,160 --> 00:55:06,600
USING NORMAL CONTROLS OR DO YOU

1647
00:55:06,600 --> 00:55:07,680
THINK THEY'RE USELESS?

1648
00:55:07,680 --> 00:55:10,080
>>NO, I THINK FOR THE

1649
00:55:10,080 --> 00:55:11,200
CO-CULTURE NORMAL CONTROLS ARE

1650
00:55:11,200 --> 00:55:11,720
VERY POWERFUL.

1651
00:55:11,720 --> 00:55:12,920
SO WE HAVE DONE THEM, NOT FOR

1652
00:55:12,920 --> 00:55:14,320
ALL THE CANCERS.

1653
00:55:14,320 --> 00:55:16,840
WE'VE ONLY DONE THEM FOR BREAST

1654
00:55:16,840 --> 00:55:18,280
AND FOR -- IN THE MOUSE CONTEXT,

1655
00:55:18,280 --> 00:55:20,840
AND IN THE HUMAN CONTEXT, WE

1656
00:55:20,840 --> 00:55:23,200
HAVE SOME KIDNEY CANCERS.

1657
00:55:23,200 --> 00:55:24,360
AND WE CAN SEE SELECTIVITY

1658
00:55:24,360 --> 00:55:26,600
BETWEEN THE TUMOR -- IF YOU

1659
00:55:26,600 --> 00:55:29,360
SELECT THEM WITH THE TUMOR, IT

1660
00:55:29,360 --> 00:55:30,200
DOESN'T KILL THE --

1661
00:55:30,200 --> 00:55:31,720
>>CAN I HAVE ONE LAST
QUESTION?

1662
00:55:31,720 --> 00:55:36,840
SORRY.

1663
00:55:36,840 --> 00:55:40,040
FOR THE TCRs WE KNOW ARE

1664
00:55:40,040 --> 00:55:42,200
NEOANTIGEN REACTIVE -- WE TRIED

1665
00:55:42,200 --> 00:55:43,720
KILLING ASSAYS BUT WE ALSO

1666
00:55:43,720 --> 00:55:46,280
TRY -- SO WE ALSO TRY TO

1667
00:55:46,280 --> 00:55:48,800
REPLICATE THAT IN A PDX MODEL.

1668
00:55:48,800 --> 00:55:50,720
ONE ISSUE WE HAD IS IMROAING

1669
00:55:50,720 --> 00:55:53,680
THOSE ORGANOIDS IN A MOUSE AS A

1670
00:55:53,680 --> 00:55:54,680
PDX MODEL, THEY HAVE BEEN

1671
00:55:54,680 --> 00:55:55,200
FAILING.

1672
00:55:55,200 --> 00:55:56,320
I DON'T KNOW IF YOU'VE

1673
00:55:56,320 --> 00:55:57,240
EXPERIENCED THAT, IF YOU HAVE

1674
00:55:57,240 --> 00:55:58,200
ANY ADVICE.

1675
00:55:58,200 --> 00:55:59,440
THE OTHER THING IS, DO YOU THINK

1676
00:55:59,440 --> 00:56:02,000
IT'S NECESSARY TO TEST IN A

1677
00:56:02,000 --> 00:56:03,600
MOUSE OR IN VITRO ASSAY --

1678
00:56:03,600 --> 00:56:05,000
>>I WOULDN'T TOUCH THE
SECOND

1679
00:56:05,000 --> 00:56:05,760
QUESTION BECAUSE THAT'S ALMOST

1680
00:56:05,760 --> 00:56:06,920
LIKE AN FDA QUESTION SO LET'S

1681
00:56:06,920 --> 00:56:08,560
NOT TALK ABOUT THAT.

1682
00:56:08,560 --> 00:56:08,840
>>RIGHT.

1683
00:56:08,840 --> 00:56:10,440
>>FOR THE FIRST ONE, YES,
IT

1684
00:56:10,440 --> 00:56:12,120
USUALLY WORKS, WE CAN TALK MORE,

1685
00:56:12,120 --> 00:56:13,080
WE CAN DISCUSS MORE.

1686
00:56:13,080 --> 00:56:14,080
BUT THERE IS PROBABLY A

1687
00:56:14,080 --> 00:56:15,240
DIFFERENCE BETWEEN THE WAY YOU

1688
00:56:15,240 --> 00:56:16,480
ENRICH THE T-CELLS AND THE WAY

1689
00:56:16,480 --> 00:56:16,880
WE DO IT.

1690
00:56:16,880 --> 00:56:17,880
SO WE CAN TALK MORE.

1691
00:56:17,880 --> 00:56:18,240
>>ALL RIGHT.

1692
00:56:18,240 --> 00:56:18,920
THANK YOU.

1693
00:56:18,920 --> 00:56:24,920
>>MORE DETAILS.

1694
00:56:24,920 --> 00:56:34,720
>>I WAS JUST WONDERING --

1695
00:56:34,720 --> 00:56:36,040
HETEROJE NATE DO YOU THINK YOU

1696
00:56:36,040 --> 00:56:37,000
OBTAIN IN YOUR CULTURES?

1697
00:56:37,000 --> 00:56:38,400
>>VERY LITTLE BECAUSE
YOU'RE

1698
00:56:38,400 --> 00:56:39,160
GETTING A BIOPSY IN THE FIRST

1699
00:56:39,160 --> 00:56:40,200
PLACE OR A SMALL SECTION OF THE

1700
00:56:40,200 --> 00:56:40,560
TUMOR.

1701
00:56:40,560 --> 00:56:42,440
SO WHATEVER YOU GET BY ITSELF IS

1702
00:56:42,440 --> 00:56:44,760
NOT REPRESENTING THE ENTIRE

1703
00:56:44,760 --> 00:56:45,200
TUMOR.

1704
00:56:45,200 --> 00:56:46,520
WHAT YOU GET ALSO, THERE IS A

1705
00:56:46,520 --> 00:56:48,000
LOT OF ATTRITION IN THE CULTURE

1706
00:56:48,000 --> 00:56:50,040
AND WHATEVER GROWS, GROWS.

1707
00:56:50,040 --> 00:56:51,440
SO WE WILL NOT BE ABLE TO ANSWER

1708
00:56:51,440 --> 00:56:53,880
THAT QUESTION IN ANY CLEAR WAY

1709
00:56:53,880 --> 00:56:54,920
BECAUSE IT IS IMPOSSIBLE TO

1710
00:56:54,920 --> 00:56:56,320
ANSWER THAT QUESTION.

1711
00:56:56,320 --> 00:56:58,160
>>DO YOU FIND IT THEN

1712
00:56:58,160 --> 00:56:59,760
SURPRISING THAT YOU GET SUCH A

1713
00:56:59,760 --> 00:57:02,360
GOOD CORRELATION BETWEEN DRUG

1714
00:57:02,360 --> 00:57:04,400
RESPONSES IN THE ORGANOIDS AND

1715
00:57:04,400 --> 00:57:06,040
WHAT WAS SEEN IN THE CLINICAL

1716
00:57:06,040 --> 00:57:06,280
SETTING?

1717
00:57:06,280 --> 00:57:07,600
>>SO YES AND NO.

1718
00:57:07,600 --> 00:57:09,480
IT IS SURPRISING THAT YOU CAN

1719
00:57:09,480 --> 00:57:11,240
SEE, BUT PERHAPS THESE ARE --

1720
00:57:11,240 --> 00:57:13,040
KEEP IN MIND THE DRUGS WE TESTED

1721
00:57:13,040 --> 00:57:14,640
ARE ALL CHEMOTHERAPY DRUGS.

1722
00:57:14,640 --> 00:57:16,840
SO IF THEY ARE SENSITIVITIES

1723
00:57:16,840 --> 00:57:18,440
DICTATED BY SOME COMMON ELEMENTS

1724
00:57:18,440 --> 00:57:20,600
THAT ARE SHAPED BY MULTIPLE

1725
00:57:20,600 --> 00:57:21,800
CELLS, MAYBE A TRUNCAL MUTATION

1726
00:57:21,800 --> 00:57:23,680
OF SOME SORT, THEN MAYBE THAT'S

1727
00:57:23,680 --> 00:57:24,960
WHAT WE ARE LOOKING AT.

1728
00:57:24,960 --> 00:57:27,000
THERE MAY BE SOME INTRATUMOR

1729
00:57:27,000 --> 00:57:27,920
DIFFERENCES IN THE WAY THEY

1730
00:57:27,920 --> 00:57:28,720
WOULD RESPOND WHEN YOU GIVE IT

1731
00:57:28,720 --> 00:57:29,840
TO THE PATIENT AND THAT IS

1732
00:57:29,840 --> 00:57:30,400
LIKELY TO BE THE CASE.

1733
00:57:30,400 --> 00:57:32,680
BUT I THINK IT IS IMPOSSIBLE TO

1734
00:57:32,680 --> 00:57:34,840
REPRESENT THE ORGANOID OR A --

1735
00:57:34,840 --> 00:57:36,920
ANY MODEL FOR THAT MATTER TO

1736
00:57:36,920 --> 00:57:40,520
CLOSELY MIMIC THE PATIENT

1737
00:57:40,520 --> 00:57:41,880
TUMOR -- ENTIRE HETEROGENEITY SO

1738
00:57:41,880 --> 00:57:43,040
THAT'S A REALITY AND WE HAVE TO

1739
00:57:43,040 --> 00:57:50,320
WORK WITH WHAT WE HAVE.

1740
00:57:50,320 --> 00:57:53,160
>>OKAY.

1741
00:57:53,160 --> 00:57:56,560
ANOTHER ONE?

1742
00:57:56,560 --> 00:57:57,560
>>HI.

1743
00:57:57,560 --> 00:57:57,880
AMAZING JOB.

1744
00:57:57,880 --> 00:57:58,880
I HAVE A QUESTION.

1745
00:57:58,880 --> 00:58:03,680
WHEN I THINK ABOUT POLARITY, I

1746
00:58:03,680 --> 00:58:06,640
ONLY THINK ABOUT CELL

1747
00:58:06,640 --> 00:58:07,200
CYTOSKELETON.

1748
00:58:07,200 --> 00:58:09,800
DID YOU HAVE ANY EVIDENCE BY

1749
00:58:09,800 --> 00:58:12,560
MODIFYING THIS POLARITY

1750
00:58:12,560 --> 00:58:13,240
PROTEIN -- IF YOU CHANGE THE

1751
00:58:13,240 --> 00:58:16,840
MORPHOLOGY OF THE CELLS LIKE

1752
00:58:16,840 --> 00:58:21,040
EVEN THE -- HOW THE CELLS MOVE?

1753
00:58:21,040 --> 00:58:23,080
>>SO IN THE CONTEXT OF
LGL2, WE

1754
00:58:23,080 --> 00:58:24,240
DID A SERIES OF EXPERIMENTS TO

1755
00:58:24,240 --> 00:58:26,720
SEE IF CELL POLARITY IS ALTERED,

1756
00:58:26,720 --> 00:58:28,920
AND WHEN WE OVEREXPRESSED LGL2,

1757
00:58:28,920 --> 00:58:30,520
WE DO NOT SEE A MAJOR CHANGE.

1758
00:58:30,520 --> 00:58:32,160
BECAUSE THERE ARE COMPENSATORY

1759
00:58:32,160 --> 00:58:32,920
MECHANISMS THAT KICK IN.

1760
00:58:32,920 --> 00:58:35,520
AND YOU DON'T SEE CHANGES IN

1761
00:58:35,520 --> 00:58:39,720
CELL-CELL JUNCTIONS OR

1762
00:58:39,720 --> 00:58:40,880
COMPOSITION OF THE --

1763
00:58:40,880 --> 00:58:41,320
CYTOSKELETON.

1764
00:58:41,320 --> 00:58:42,560
IF YOU KNOCK DOWN, THERE IS A

1765
00:58:42,560 --> 00:58:45,920
CHANCE YOU CHANGE YOU WILL SEE E

1766
00:58:45,920 --> 00:58:47,000
MORPHOLOGY, BUT IF YOU LOOK

1767
00:58:47,000 --> 00:58:48,400
AT -- JUNCTIONS AND SO FORTH,

1768
00:58:48,400 --> 00:58:49,600
IT'S NOT DRAMA IK IT ENOUGH.

1769
00:58:49,600 --> 00:58:51,000
THE GROWTH RATES ARE THE SAME,

1770
00:58:51,000 --> 00:58:52,800
THE STRUCTURE IS MORE OR LESS

1771
00:58:52,800 --> 00:58:53,080
CONSERVED.

1772
00:58:53,080 --> 00:58:54,480
IN PART THAT'S BECAUSE POLARITY

1773
00:58:54,480 --> 00:58:56,760
PROTEINS HAVE A HUGE -- PAUSE

1774
00:58:56,760 --> 00:58:58,080
YOU HAVE TO MAINTAIN THE

1775
00:58:58,080 --> 00:58:58,920
STRUCTURE AT ALL COSTS.

1776
00:58:58,920 --> 00:59:00,800
SO IT'S RARELY THAT IN A

1777
00:59:00,800 --> 00:59:05,920
MAMMALIAN CELL, YOU RUN A

1778
00:59:05,920 --> 00:59:07,000
DOMINANT PHENOTYPE -- YOU HAVE

1779
00:59:07,000 --> 00:59:08,240
TO TAKE OUT A CLASS, FOR

1780
00:59:08,240 --> 00:59:11,920
EXAMPLE.

1781
00:59:11,920 --> 00:59:14,600
>>ANY OTHER QUESTIONS?

1782
00:59:14,600 --> 00:59:14,800
OKAY.

1783
00:59:14,800 --> 00:59:15,160
>>THANK YOU.

1784
00:59:15,160 --> 00:59:21,160
[APPLAUSE]