Cargando…

Development of a multiplex assay to assess activated p300/CBP in circulating prostate tumor cells

Reduced SIRT2 deacetylation and increased p300 acetylation activity leads to a concerted mechanism of hyperacetylation at specific histone lysine sites (H3K9, H3K14, and H3K18) in castration-resistant prostate cancer (CRPC). We examined whether circulating tumor cells (CTCs) identify patients with a...

Descripción completa

Detalles Bibliográficos
Autores principales: Filon, Mikolaj, Yang, Bing, Purohit, Tanaya A., Schehr, Jennifer, Singh, Anupama, Bigarella, Marcelo, Lewis, Peter, Denu, John, Lang, Joshua, Jarrard, David F.
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Impact Journals LLC 2023
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10360924/
https://www.ncbi.nlm.nih.gov/pubmed/37477521
http://dx.doi.org/10.18632/oncotarget.28477
_version_ 1785076120508432384
author Filon, Mikolaj
Yang, Bing
Purohit, Tanaya A.
Schehr, Jennifer
Singh, Anupama
Bigarella, Marcelo
Lewis, Peter
Denu, John
Lang, Joshua
Jarrard, David F.
author_facet Filon, Mikolaj
Yang, Bing
Purohit, Tanaya A.
Schehr, Jennifer
Singh, Anupama
Bigarella, Marcelo
Lewis, Peter
Denu, John
Lang, Joshua
Jarrard, David F.
author_sort Filon, Mikolaj
collection PubMed
description Reduced SIRT2 deacetylation and increased p300 acetylation activity leads to a concerted mechanism of hyperacetylation at specific histone lysine sites (H3K9, H3K14, and H3K18) in castration-resistant prostate cancer (CRPC). We examined whether circulating tumor cells (CTCs) identify patients with altered p300/CBP acetylation. CTCs were isolated from 13 advanced PC patients using Exclusion-based Sample Preparation (ESP) technology. Bound cells underwent immunofluorescent staining for histone modifying enzymes (HMEs) of interest and image capture with NIS-Elements software. Using the cBioPortal PCF/SU2C dataset, the response of CRPC to androgen receptor signaling inhibitors (ARSI) was analyzed in 50 subjects. Staining optimization and specificity revealed clear expression of acetyl-p300, acetyl-H3K18, and SIRT2 on CTCs (CK positive, CD45 negative cells). Exposure to A-485, a selective p300/CBP catalytic inhibitor, reduced p300 and H3K18 acetylation. In CRPC patients, a-p300 strongly correlated with its target acetylated H3k18 (Pearson’s R = 0.61), and SIRT2 expression showed robust negative correlation with a-H3k18 (R = −0.60). A subgroup of CRPC patients (6/11; 55%) demonstrated consistent upregulation of acetylation based on these markers. To examine the clinical impact of upregulation of the CBP/p300 axis, CRPC patients with reduced deacetylase SIRT2 expression demonstrate shorter response times to ARSI therapy (5.9 vs. 12 mo; p = 0.03). A subset of CRPC patients demonstrate increased p300/CBP activity based on a novel CTC biomarker assay. With further development, this biomarker suite may be used to identify candidates for CBP/p300 acetylation inhibitors in clinical development.
format Online
Article
Text
id pubmed-10360924
institution National Center for Biotechnology Information
language English
publishDate 2023
publisher Impact Journals LLC
record_format MEDLINE/PubMed
spelling pubmed-103609242023-07-22 Development of a multiplex assay to assess activated p300/CBP in circulating prostate tumor cells Filon, Mikolaj Yang, Bing Purohit, Tanaya A. Schehr, Jennifer Singh, Anupama Bigarella, Marcelo Lewis, Peter Denu, John Lang, Joshua Jarrard, David F. Oncotarget Research Paper Reduced SIRT2 deacetylation and increased p300 acetylation activity leads to a concerted mechanism of hyperacetylation at specific histone lysine sites (H3K9, H3K14, and H3K18) in castration-resistant prostate cancer (CRPC). We examined whether circulating tumor cells (CTCs) identify patients with altered p300/CBP acetylation. CTCs were isolated from 13 advanced PC patients using Exclusion-based Sample Preparation (ESP) technology. Bound cells underwent immunofluorescent staining for histone modifying enzymes (HMEs) of interest and image capture with NIS-Elements software. Using the cBioPortal PCF/SU2C dataset, the response of CRPC to androgen receptor signaling inhibitors (ARSI) was analyzed in 50 subjects. Staining optimization and specificity revealed clear expression of acetyl-p300, acetyl-H3K18, and SIRT2 on CTCs (CK positive, CD45 negative cells). Exposure to A-485, a selective p300/CBP catalytic inhibitor, reduced p300 and H3K18 acetylation. In CRPC patients, a-p300 strongly correlated with its target acetylated H3k18 (Pearson’s R = 0.61), and SIRT2 expression showed robust negative correlation with a-H3k18 (R = −0.60). A subgroup of CRPC patients (6/11; 55%) demonstrated consistent upregulation of acetylation based on these markers. To examine the clinical impact of upregulation of the CBP/p300 axis, CRPC patients with reduced deacetylase SIRT2 expression demonstrate shorter response times to ARSI therapy (5.9 vs. 12 mo; p = 0.03). A subset of CRPC patients demonstrate increased p300/CBP activity based on a novel CTC biomarker assay. With further development, this biomarker suite may be used to identify candidates for CBP/p300 acetylation inhibitors in clinical development. Impact Journals LLC 2023-07-20 /pmc/articles/PMC10360924/ /pubmed/37477521 http://dx.doi.org/10.18632/oncotarget.28477 Text en Copyright: © 2023 Filon et al. https://creativecommons.org/licenses/by/3.0/This is an open access article distributed under the terms of the Creative Commons Attribution License (https://creativecommons.org/licenses/by/3.0/) (CC BY 3.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.
spellingShingle Research Paper
Filon, Mikolaj
Yang, Bing
Purohit, Tanaya A.
Schehr, Jennifer
Singh, Anupama
Bigarella, Marcelo
Lewis, Peter
Denu, John
Lang, Joshua
Jarrard, David F.
Development of a multiplex assay to assess activated p300/CBP in circulating prostate tumor cells
title Development of a multiplex assay to assess activated p300/CBP in circulating prostate tumor cells
title_full Development of a multiplex assay to assess activated p300/CBP in circulating prostate tumor cells
title_fullStr Development of a multiplex assay to assess activated p300/CBP in circulating prostate tumor cells
title_full_unstemmed Development of a multiplex assay to assess activated p300/CBP in circulating prostate tumor cells
title_short Development of a multiplex assay to assess activated p300/CBP in circulating prostate tumor cells
title_sort development of a multiplex assay to assess activated p300/cbp in circulating prostate tumor cells
topic Research Paper
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10360924/
https://www.ncbi.nlm.nih.gov/pubmed/37477521
http://dx.doi.org/10.18632/oncotarget.28477
work_keys_str_mv AT filonmikolaj developmentofamultiplexassaytoassessactivatedp300cbpincirculatingprostatetumorcells
AT yangbing developmentofamultiplexassaytoassessactivatedp300cbpincirculatingprostatetumorcells
AT purohittanayaa developmentofamultiplexassaytoassessactivatedp300cbpincirculatingprostatetumorcells
AT schehrjennifer developmentofamultiplexassaytoassessactivatedp300cbpincirculatingprostatetumorcells
AT singhanupama developmentofamultiplexassaytoassessactivatedp300cbpincirculatingprostatetumorcells
AT bigarellamarcelo developmentofamultiplexassaytoassessactivatedp300cbpincirculatingprostatetumorcells
AT lewispeter developmentofamultiplexassaytoassessactivatedp300cbpincirculatingprostatetumorcells
AT denujohn developmentofamultiplexassaytoassessactivatedp300cbpincirculatingprostatetumorcells
AT langjoshua developmentofamultiplexassaytoassessactivatedp300cbpincirculatingprostatetumorcells
AT jarrarddavidf developmentofamultiplexassaytoassessactivatedp300cbpincirculatingprostatetumorcells