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Induction of PD-L1 expression by forced ASS1 overexpression was accompanied by an increase in interferon type I signaling.

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Similar results were obtained in a mesothelioma cell line developing resistance to ADI-PEG20 under long-term culture.

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Next, ADI-PEG20 treatment triggered release of interferon-alpha/beta which induced PD-L1 expression by 24hrs in the ASS1-negative MPM cell lines before declining by 48hrs.

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¡´³o­Ó³¡¤ÀADI+KeytrudaÀ³¸Ó´N¬O³o¼Ë°µªº¡÷ADI-PEG 20 (36 mg/m2) weekly in combination with pembrolizumab (1 and 2 mg/kg or 200 mg) every three weeks¡÷²Ä¤@¤Ñ¥´ADI¡÷²Ä2 ©Î 3¤Ñ¥´Keytruda¡÷²Ä8,15,22¤Ñ¦A¥´ADI¡÷²Ä23 or 24¤Ñ¥´Keytruda.......

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The primary endpoint for the phase 2 stage will be overall response rate (ORR) with secondary endpoints of overall survival (OS), safety and toxicity. The phase 2 will test ORR proportions with the placebo triplet set at 15% vs. 35% for the ADI-PEG 20 triplet, with a 1:1 randomization, 80% power.

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¦¨¥\¤£¥²¦b§Ú¡A¨ÖÁʤj¨Æ¥ó¥²±Nµo¥Í¡A°ß¦³¤Q¤j¼t¤è¯à¹ê²{§d³ÕªºÄ@±æ¡A

ÅýÀù¯g¦¨¬°ºC©Ê¯f¡C

¹ï©óºô¤W¬Y¨Ç¤H¡A¬°¤F¤p§Q¡A¹H­I¨}¤ß»¡¸Ü¡A¬ÝµÛ¦³®É«Ü¥Í®ð¡A³o¬O§Ú­×¾i¤£¨¬¡C

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¤Ñ©R¤j¡A«Ü´Î¡I³Ìªñ³£¨S¦³¬Ý¨ì§A¡A­ì¨Ó§A¦b¥Î¥\Ū¦Ñ´­ªººëµØ¡Aµª®×¦b¾ú¦¸ªº¤å³¹¤¤³£¤w³°Äò¥æ«Ý¡A­Y«ÜÃø§ä©Î«ÜÃøÀ´¡A±ß¤W¦³ªÅ¦A¦^µª§A¡C­n¥D«ù±á·|¤F¡C¥[ªo¡A§A¶V¨Ó¶V´Î¤F¡C
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1. ¼Ë¥»¼Æn=225¤v©w¡A­n¦A¼W¥[°£«DFDAÂIÀY¡A¤£µM¬O¨S¦³¿ìªk¦Û¤v¨M©w¼W¥[ªº¡C

2. 4/14=28.57%¡A¦ý¦]¼Ë¥»¼Æ¤Ó¤Ö¡AÀË©w¤OPower¥u¦³0.39(39%)¡÷´N¬O»¡·Ó³o¼Ë¥h±À®i¨ì¥þ¤HÃþ®É¡A¥u¦³39%ªº¾÷·|¸ò³o¦¸ªºµª®×¤@¼Ë¡I

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3. Single arm, open label¨S¦³´Á¤¤¤ÀªR(interim analysis)ªº°ÝÃD¡I¥i¥HÀH®É¤ÀªR¨Ó½Õ¾ã¡I

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¸ò¾Ú¤ÀªRµ²ªG½Õ¾ã(adaptive)¸ÕÅç³]­p(¥D­n¬Osample size)¡A³o¯A¤ÎType I errorªº±±¨î¡C

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FDA guidance¡G Adaptive Design Clinical Trials for Drugs and Biologics www.fda.gov/downloads/Drugs/GuidanceComplianceRegulatoryInformation/Guidances/UCM201790.pdf

¥Ñ©óAdaptive Study Design For Single-Arm Trials§xÃø«Ü¦h¡A½Ð²Î­p±M®a³]­p§¹«á¡A³Ì¦n¦A°Ý°ÝFDAªº·N¨£¡C

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Â^¨ú¤@¨Ç­«­nªº¬q¸¨¤À¨É

Three ASS1-negative and one ASS1-positive MPM cell lines were assessed for PD-L1 expression by real-time quantitative PCR, western blot and FACS analysis.

¡´¥ÎPCR, western blot and FACS¤ÀªR3­ÓASS1-negative & 3­ÓASS1- positiveªºªÍ¶¡¥ÖÀù(MPM)²Ó­M®è(cell lines)¡C

An immunocompetent murine tumor model of ASS1 loss mimicking aggressive mesothelioma was treated with PBS control, ADI-PEG20, anti-PD-1 antibody, and ADI-PEG20 plus anti-PD-1 antibody.

¡´¼Ò¥é¦³§K¬Ì¬¡©Ê¥B¯Ê¥FASS1ªº¹«Ãþ«I²¤©ÊMPM¸~½F¼Ò«¬¡A¥ÎPBS control, ADI-PEG20, anti-PD-1 antibody, and ADI-PEG20 plus anti-PD-1 antibody¥|ºØ¤è¦¡¥h´ú¸Õ¡C

Finally, human mesothelioma biopsies from trials of ADI-PEG20 were analyzed for ASS1 and PD-L1 and immune cell subsets.

¡´¥Î·íªìMPM¥HADIªvÀøªº¯f¤Hªº¸~½F¤Á¤ù¼Ë¥»¨Ó¤ÀªR¡AASS1, PD-L1 and immune cell subsets.

µ²ªGµo²{

PDL1 protein was absent in the three ASS1 negative MPM cell lines but was present in the ASS1 positive cell line.

¡´3­ÓASS1-negativeªºMPM²Ó­M®è¬O¯Ê¥FPD-L1ªº¡F¦ý¬O1­ÓASS1- positiveªºMPM²Ó­M®è¬O¦³PD-L1ªº¡I

Transfection of ASS1 in the ASS1 negative MPM cell lines led to an increase in PD-L1 expression, which was reversible following ASS1 knockdown.

¡´§âASS1Âà¥[¨ìASS1-negativeªºMPM²Ó­M®èµo²{¡APD-L1 expression¤]¸òµÛ¤W¤É¡A³o­Ó¾÷¨î¬O¥i°fªº¡A·íASS1¤U­°®ÉPD-L1 expression¤]¸òµÛ¤U­°¤F¡I

Induction of PD-L1 expression by forced ASS1 overexpression was accompanied by an increase in interferon type I signaling.

¡´±j­¢ASS1¹L«×ªí¹F¨ÏPD-L1 expression¤]¸òµÛ¤W¤É®É¡Ainterferon type I signaling¤]¦ñÀH¤W¤É¡C

Similar results were obtained in a mesothelioma cell line developing resistance to ADI-PEG20 under long-term culture.

¡´MPM²Ó­M®è¤]¬O¦P¼Ëªºµ²ªG¡A¦bªø´Á°ö¾i¤Uµo®i¦¨¹ïADI¦³§Ü©Ê¡C

Next, ADI-PEG20 treatment triggered release of interferon-alpha/beta which induced PD-L1 expression by 24hrs in the ASS1-negative MPM cell lines before declining by 48hrs.

¡´ADIªºªvÀø¥i¿Eµointerferon- alpha/betaªºÄÀ©ñ¡Ainterferon- alpha/betaªºÄÀ©ñ¥i¦b24¤p®É¤º(¦b¤U­°«e-48¤p®É)»¤¾ÉASS1-negative ªºMPM²Ó­M®èªºPD-L1 expression¡A

Analysis of MPM biopsies of patients progressing on ADI-PEG20 revealed upregulation of ASS1 and a concomitant increase in tumoral PD-L1 and CD3 positive T cells.

¡´¥Î±µ¨ü¹LADIªvÀø«á´_µoªºMPM¯f¤Hªº¸~½F¤Á¤ù¤ÀªRµo²{¡AASS1¤W¤É·|¦ñÀHPD-L1ªºªí²{©MCD3 positive T cell¤W¤É¡C

ADI-PEG20 synergized with PD-1 blockade in the immunocompetent murine tumor model that was refractory to PD-1 inhibition.

¡´©Ò¥H¡AADI©Manti-PD1¦b¹«¬ì°Êª«(§Y«K¬O¹ï§í¨îPD-1¤w¦³­@¨ü©Ê)¬O¦³¨ó¦P¥[¦¨ªºÀø®Äªº¡I

µ²½×

ASS1 and ADI-PEG20 modulate PDL1 expression via type I interferon signaling in malignant mesothelioma cell lines.

¡´ASS1 and ADI¥i¥H³z¹Ltype I interferon signaling½Õ±±PD-L1ªºªí²{

Arginine deprivation with ADI-PEG20 combined with PD-1 blockade is synergistic and warrants further exploration in the clinic.

¡´ADI+ anti-PD-1¬O¦³¨ó¦P¥[¦¨ªºÀø®Ä¡A©|»ÝÁ{§É¬ã¨s¨Ó±´°QÃÒ¹ê

A phase 1 trial of ADI-PEG20 combined with PD1 blockade is planned in patients with ASS1-negative cancers, including malignant mesothelioma.

¡´°w¹ïASS1-negativeªºÀù§O¡A Phase 1ªºADI+anti-PD1¤v¸g¦b¶i¦æ(¥]¬AMPM)¡I

¦Ñ´­ÂIµû¡G

±qcell line¡÷°Êª«¼Ë¥»¼Ò«¬¤w¸g¬Ý¨ìADI+ anti-PD-1ªº¨ó¦P¥[¦¨Àø®Ä

¡´ °w¹ïASS1-negative¸û¦hªºÀù§O(¦]¬°PD-L1 expression¤]¤Ö)--¾A¥ÎADI¡AªvÀø>8~15¶gºëÓi»Ä­°¸Ñ¤U­°®É¡AASS1-positive¤W¤É¡A³o­Ó®É­ÔPD-L1 expression¤]¤W¤É¡A­è¦nanti-PD1¬£¤W¥Î³õ¡I

¡´°w¹ïASS1-positive¸û¦hªºÀù§O(PD-L1 expression¤]¦h)--¾A¥Îanti-PD1¡AªvÀø¤@¬q®É¶¡PD-L1 expression¤U­°®É¡AASS1-positive¤]¤U­°¡A³o®ÉADI¤S¥i¥Hµo´§§ó¤jªº§@¥Î¡I

¡´¨âªÌ¬Û»²¬Û¦¨¡A»P°ê½Ã°|¦X§@ªºADI+Keytruda¥Ø«e¦¬6¦ì,4¦ì­è¦¬,¨ä¥L2¦ìªº®ÄªG³£«D±`¦n¡ANSCLC 1¦ì, H&NSCC 1¦ì¡I¤½¥q¹w­p©ú¦~ASCO±N´¦ÅS³o¨Ç¼Æ¾Ú¡I·íµM¦b¥¼¨Óªº3­Ó¤ë¤º´N·|¦³¤wñ«O±K¨ó©wªº¤jÃļt¬Ý¹L³o¨Ç¼Æ¾Ú¡I©¡®É¡A¾_¾Ù±NçE¤Ñ»\¦a¡I

¡´¥H¥_·¥¬Pªº¬ãµo¯à¤O¬ãµo¥Xªº±M§Q¡BÃÄ«~©M¼t¦a¡A´Nºâ¯}²£½â½æÀ³¸Ó¤]¥i¥H½æ­Ó10E~20E¬üª÷¡I---¦Û¤v´«ºâ¤@¤U¡I¦pªG¥u¹ÏÁÈ­Ó50%~100%§Q¼íªº§ë¸êªÌ¡A§A¬Æ¦Ü¥i¥H¬èë¥_·¥¬P»°§Ö¯}²£¡I·|½æªº§ó§Ö¤@ÂI¡I

¡´¨p¶Ò´Ú¦¬»ô·|¦b10/06 or 10/11¤½§i¡A·Q·Q18¶ô¼W¸ê©M33.6¶ô¨p¶Ò¤½§i

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Result:

PDL1 protein was absent in the three ASS1 negative MPM cell lines but was present in the ASS1 positive cell line. Transfection of ASS1 in the ASS1 negative MPM cell lines led to an increase in PD-L1 expression, which was reversible following ASS1 knockdown. Induction of PD-L1 expression by forced ASS1 overexpression was accompanied by an increase in interferon type I signaling. Similar results were obtained in a mesothelioma cell line developing resistance to ADI-PEG20 under long-term culture. Next, ADI-PEG20 treatment triggered release of interferon-alpha/beta which induced PD-L1 expression by 24hrs in the ASS1-negative MPM cell lines before declining by 48hrs. Analysis of MPM biopsies of patients progressing on ADI-PEG20 revealed upregulation of ASS1 and a concomitant increase in tumoral PD-L1 and CD3 positive T cells. ADI-PEG20 synergized with PD-1 blockade in the immunocompetent murine tumor model that was refractory to PD-1 inhibition.

Conclusion:

ASS1 and ADI-PEG20 modulate PDL1 expression via type I interferon signaling in malignant mesothelioma cell lines. Arginine deprivation with ADI-PEG20 combined with PD-1 blockade is synergistic and warrants further exploration in the clinic. A phase 1 trial of ADI-PEG20 combined with PD1 blockade is planned in patients with ASS1-negative cancers, including malignant mesothelioma.

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Dr. Peter Szlosarek°w¹ïPegylated Arginine Deiminase Potentiates PD-1/PD-L1 Immune Checkpoint Blockade in Malignant Mesothelioma

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MA 19.05 - Pegylated Arginine Deiminase Potentiates PD-1/PD-L1 Immune Checkpoint Blockade in Malignant Mesothelioma (ID 9207)

11:30 - 11:35 AM | Presenting Author(s): Peter Szlosarek | Author(s): R. Khadeir, M. Sheaff, M. Locke, K. Lau, B. Wu, J. Bomalaski, S. Martin, S. Quezada

 Abstract

The abstract for this presentation will be made available at 21:00 UTC on September 27, 2017 (14:00 Pacific Daylight Time). Please return after this date to review more information about this presentation.

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¤¤°ê-«H°U 65(¶R) 2645(½æ) -2580(½æ¶W) 92.11(§¡»ù)= 2.37E

¤¸¤j-·s¦Ë 25(¶R) 1882(½æ) -1856(½æ¶W) 90.48(§¡»ù)= 1.68E

¤¸¤j-ªO¾ô 15(¶R) 195(½æ) -180(½æ¶W) 91.52(§¡»ù)= 0.16E

³Í°ò-¥_ªù 2(¶R) 96(½æ) -94(½æ¶W) 88.58(§¡»ù)= 0.08E

106/09/25~106/09/26

¤¤°ê-«H°U 113(¶R) 243(½æ) -130(½æ¶W) 91.98(§¡»ù)= 0.12E

¤¸¤j-·s¦Ë 5(¶R) 223(½æ) -218(½æ¶W) 92.07(§¡»ù)= 0.2E

³Í°ò-¥_ªù 1(¶R) 296(½æ) -295(½æ¶W) 92.92(§¡»ù)= 0.27E

­P©M-«n¨Ê 0(¶R) 406(½æ) -406(½æ¶W) 88.70(§¡»ù)= 0.36E

106/09/27

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­P©M-«n¨Ê 0(¶R) 256(½æ) -256(½æ¶W) 81.38(§¡»ù)= 0.21E

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