TY - JOUR
T1 - Small-molecule natural product physachenolide C potentiates immunotherapy efficacy by targeting BET proteins
AU - Tewary, Poonam
AU - Brooks, Alan D.
AU - Xu, Ya Ming
AU - Kithsiri Wijeratne, E. M.
AU - Babyak, Ashley L.
AU - Back, Timothy C.
AU - Chari, Raj
AU - Evans, Christine N.
AU - Henrich, Curtis J.
AU - Meyer, Thomas J.
AU - Edmondson, Elijah F.
AU - De Aquino, Maria T.Prudente
AU - Kanagasabai, Thanigaivelan
AU - Shanker, Anil
AU - Leslie Gunatilaka, A. A.
AU - Sayers, Thomas J.
N1 - Funding Information:
This study was funded in whole or in part with federal funds from the NCI, NIH, under contract HHSN26120080001E (awarded to T.J. Sayers). The content of this publication does not necessarily reflect the views or policies of the Department of Health and Human Services, nor does mention of trade names, commercial products or organizations imply endorsement by the U.S. Government. This work was also supported by a grant from the Arizona Biomedical Research Center (grant number ADHS-16-162515; awarded to A.A.L. Gunatilaka) and (in part) by the Intramural Research Program of NIH, Frederick National Lab, Cancer and Inflammation program (CIP), Center for Cancer Research (CCR), and by funds from the NIH grants (grant numbers U54 CA163069 and SCI CA182843; awarded to A. Shanker). The authors thank Dr. Julio Valencia (CIP, NCI Frederick) for his scientific suggestions, discussions, and valuable comments, Ms. Anna Trivett for her technical assistance, Ms. Roberta Matthai for FACS single-cell sorting of cells, Ms. Nirmala Sharma for her assistance with cFLIP blots, and Ms. Megan Karwan for her assistance with in vivo studies. They would also like to thank Drs. Maggie Cam and Jack Chen (CCR Collaborative Bioinformatics Resource, NCI) and Steven Anderson (CIP, NCI Frederick) for their help with the analysis of RNA-sequencing data, Drs. Thorkell Andresson and Sudipto Das (Protein Characterization Lab, FNL) for their assistance in themass spectrometric identification of cellular target proteins. Mass spectrometric and proteomics data for identification of cellular target proteins were also acquired by Drs. Krishna Parsawar and Cynthia David of the University of Arizona Proteomics Consortium (supported by NIEHS grant ES06694 to the SWEHSC, NIH/NCI grant CA023074 to the UA Cancer Center and by the BIO5 Institute of the University of Arizona). The authors are also grateful to Drs. Marc Hennequart and Benoit van den Eynde, Ludwig Cancer Institute, Brussels, Belgium, for their helpful advice on adoptive cell transfer in NSGmice, Dr. Esteban Celis (Georgia Cancer Center, GA) for his critical reading of the manuscript, and Andrew Sayers for his assistance with the artwork.
Funding Information:
This study was funded in whole or in part with federal funds from the NCI, NIH, under contract HHSN26120080001E (awarded to T.J. Sayers). The content of this publication does not necessarily reflect the views or policies of the Department of Health and Human Services, nor does mention of trade names, commercial products or organizations imply endorsement by the U.S. Government. This work was also supported by a grant from the Arizona Biomedical Research Center (grant number ADHS-16–162515; awarded to A.A.L. Gunatilaka) and (in part) by the Intramural Research Program of NIH, Frederick National Lab, Cancer and Inflammation program (CIP), Center for Cancer Research (CCR), and by funds from the NIH grants (grant numbers U54 CA163069 and SCI CA182843; awarded to A. Shanker). The authors thank Dr. Julio Valencia (CIP, NCI Frederick) for his scientific suggestions, discussions, and valuable comments, Ms. Anna Trivett for her technical assistance, Ms. Roberta Matthai for FACS single-cell sorting of cells, Ms. Nirmala Sharma for her assistance with cFLIP blots, and Ms. Megan Karwan for her assistance with in vivo studies. They would also like to thank Drs. Maggie Cam and Jack Chen (CCR Collaborative Bioinformatics Resource, NCI) and Steven Anderson (CIP, NCI Frederick) for their help with the analysis of RNA-sequencing data, Drs. Thorkell Andresson and Sudipto Das (Protein Characterization Lab, FNL) for their assistance in the mass spectrometric identification of cellular target proteins. Mass spectrometric and proteomics data for identification of cellular target proteins were also acquired by Drs. Krishna Parsawar and Cynthia David of the University of Arizona Proteomics Consortium (supported by NIEHS grant ES06694 to the SWEHSC, NIH/NCI grant CA023074 to the UA Cancer Center and by the BIO5 Institute of the University of Arizona). The authors are also grateful to Drs. Marc Hennequart and Benoit van den Eynde, Ludwig Cancer Institute, Brussels, Belgium, for their helpful advice on adoptive cell transfer in NSG mice, Dr. Esteban Celis (Georgia Cancer Center, GA) for his critical reading of the manuscript, and Andrew Sayers for his assistance with the artwork.
Funding Information:
P. Tewary reports a patent for 10849912 issued and a patent for US20210047363A1 pending. A.D. Brooks reports a patent for 10849912 issued and a patent for US20210047363A1 pending. Y. Xu reports a patent 10849912 issued, a patent US20210047363A1 pending, and a patent 20110230551 pending. E. Wijeratne reports a patent for 10849912 issued and a patent for US20210047363A1 pending. C.J. Henrich reports a patent for 10849912 issued. A. Gunatilaka reports grants from Arizona Biomedical Research Center during the conduct of the study; other support from Sun Pharma Advanced Research Co. Ltd., India and grants from Regulonix, LLC., outside the submitted work; in addition, A. Gunatilaka has a patent 10849912 issued and a patent for US20210047363A1 pending. T.J. Sayers reports a patent for
Publisher Copyright:
© 2021 American Association for Cancer Research.
PY - 2021/6
Y1 - 2021/6
N2 - Screening for sensitizers of cancer cells to TRAIL-mediated apoptosis identified a natural product of the 17β-hydroxywithanolide (17-BHW) class, physachenolide C (PCC), as a promising hit. In this study, we show that PCC was also able to sensitize melanoma and renal carcinoma cells to apoptosis in response not only to TRAIL, but also to the synthetic polynucleotide poly I:C, a viral mimetic and immune activator, by reducing levels of antiapoptotic proteins cFLIP and Livin. Both death receptor and TLR3 signaling elicited subsequent increased assembly of a proapoptotic ripoptosome signaling complex. Administration of a combination of PCC and poly I:C in human M14 melanoma xenograft and a syngeneic B16 melanoma model provided significant therapeutic benefit as compared with individual agents. In addition, PCC enhanced melanoma cell death in response to activated human T cells in vitro and in vivo in a death ligand-dependent manner. Biochemical mechanism-ofaction studies established bromo and extraterminal domain (BET) proteins as major cellular targets of PCC. Thus, by targeting of BET proteins to reduce antiapoptotic proteins and enhance caspase-8-dependent apoptosis of cancer cells, PCC represents a unique agent that can potentially be used in combination with various immunotherapeutic approaches to promote tumor regression and improve outcome.
AB - Screening for sensitizers of cancer cells to TRAIL-mediated apoptosis identified a natural product of the 17β-hydroxywithanolide (17-BHW) class, physachenolide C (PCC), as a promising hit. In this study, we show that PCC was also able to sensitize melanoma and renal carcinoma cells to apoptosis in response not only to TRAIL, but also to the synthetic polynucleotide poly I:C, a viral mimetic and immune activator, by reducing levels of antiapoptotic proteins cFLIP and Livin. Both death receptor and TLR3 signaling elicited subsequent increased assembly of a proapoptotic ripoptosome signaling complex. Administration of a combination of PCC and poly I:C in human M14 melanoma xenograft and a syngeneic B16 melanoma model provided significant therapeutic benefit as compared with individual agents. In addition, PCC enhanced melanoma cell death in response to activated human T cells in vitro and in vivo in a death ligand-dependent manner. Biochemical mechanism-ofaction studies established bromo and extraterminal domain (BET) proteins as major cellular targets of PCC. Thus, by targeting of BET proteins to reduce antiapoptotic proteins and enhance caspase-8-dependent apoptosis of cancer cells, PCC represents a unique agent that can potentially be used in combination with various immunotherapeutic approaches to promote tumor regression and improve outcome.
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UR - http://www.scopus.com/inward/citedby.url?scp=85108080992&partnerID=8YFLogxK
U2 - 10.1158/0008-5472.CAN-20-2634
DO - 10.1158/0008-5472.CAN-20-2634
M3 - Article
C2 - 33837043
AN - SCOPUS:85108080992
VL - 81
SP - 3374
EP - 3386
JO - Cancer Research
JF - Cancer Research
SN - 0008-5472
IS - 12
ER -