For over half a century, prostate cancer research has focused on the Androgen receptor (AR), its role in the initiation of the disease and progression to the highly metastatic stage, castration resistant prostate cancer (CRPC). While AR antagonists such as enzalutamide or androgen- synthesis inhibitor abiraterone dampen AR functional activity, these are often ineffective long term, as the recalcitrant disease recurs within 2-3 years. CRPCs not only thrive under low or castrate levels of androgen, but also fastidiously maintain functional AR. Consequently, anti- androgen-resistance has become one of the most vexing problems in prostate cancer therapy. Despite intensive efforts, targeting co-factors that regulate AR and its splice variant, AR-V7 transcription, directly and efficaciously, with small molecule inhibitors has not yet been achieved. We uncovered that AR recruits the oncogene ACK1, a non-receptor tyrosine kinase, to regulate its own expression- setting up a pathological positive feedback loop in androgen deprived condition. Mechanistically, activated ACK1 modifies chromatin via phosphorylation of the histone H4 at a novel site, tyrosine 88 (pY88-H4), upstream of the AR gene at three sites AREM1-3, to promote transcription. Conversely, reversal of this pY88-H4 histone modification by treatment with a novel ACK1 inhibitor, (R)-9bMS, or overexpression of the H4Y88F mutant significantly suppressed transcription of the full length AR as well as AR-V7 splice variant, consequently downregulating expression of AR-target genes, such as PSA. Moreover, we demonstrate that WDR5/MLL2 histone-Lysine N-Methyltransferase complex, interact with the pY88-H4 epigenetic marks, deposit the transcriptionally activating H3K4 tri-methyl marks in trans, thus uncovering a novel mode of epigenetic regulation at the AR locus. Based on these extensive preliminary data, we hypothesize that inhibition of ACK1 epigenetic regulator activity that suppresses AR transcription and block production of AR splice variants will be critical to combat enzalutamide- and abiraterone-resistant CRPCs. To address this hypothesis, we will pursue the following aims:
Aim 1. Assess the functional role of pY88-H4 deposition sites AREM1-3 on AR/AR-V7 transcription in CRPCs.
Aim 2. Examine genome wide MLL2/WDR5/ASH2L and pY88-H4 co-association and its role in driving AR/AR-V7 expression and CRPC progression.
Aim 3. Detail characterization of ACK1 inhibitor (R)-9bMS to overcome enzalutamide & abiraterone-resistance and ADME/DMPK studies.

Public Health Relevance

Presence of elevated AR mRNA levels in spite of prolonged androgen receptor (AR) antagonist treatment in castration resistant prostate cancer (CRPC) is a paradox that has mystified researchers. This proposal uncovers a novel mechanism of auto- regulation of AR transcription wherein AR protein coordinates functionally with ACK1 tyrosine kinase to modify chromatin within the AR gene. Significantly, these data have exposed an Achilles' heel for targeting new treatment strategies in CRPCs. Our preliminary data indicates that reversal of these epigenetic marks by a novel ACK1 inhibitor (R)-9bMS, that we have developed, opens up a new therapeutic option for this essentially incurable malignancy. This proposal will perform detailed mechanistic & in vivo studies to test efficacy of (R)-9b as novel epigenetic inhibitors that suppress both AR and AR-V7 transcription overcoming CRPC tumor growth.

Agency
National Institute of Health (NIH)
Institute
National Cancer Institute (NCI)
Type
Research Project (R01)
Project #
7R01CA208258-03
Application #
9843333
Study Section
Basic Mechanisms of Cancer Therapeutics Study Section (BMCT)
Program Officer
Sathyamoorthy, Neeraja
Project Start
2017-03-20
Project End
2022-02-28
Budget Start
2019-03-01
Budget End
2020-02-29
Support Year
3
Fiscal Year
2019
Total Cost
Indirect Cost
Name
Washington University
Department
Surgery
Type
Schools of Medicine
DUNS #
068552207
City
Saint Louis
State
MO
Country
United States
Zip Code
63130
Mahajan, Nupam P; Coppola, Domenico; Kim, Jongphil et al. (2018) Blockade of ACK1/TNK2 To Squelch the Survival of Prostate Cancer Stem-like Cells. Sci Rep 8:1954
Mahajan, Nupam P; Malla, Pavani; Bhagwat, Shambhavi et al. (2017) WEE1 epigenetically modulates 5-hmC levels by pY37-H2B dependent regulation of IDH2 gene expression. Oncotarget 8:106352-106368
Wu, Xinyan; Zahari, Muhammad Saddiq; Renuse, Santosh et al. (2017) The non-receptor tyrosine kinase TNK2/ACK1 is a novel therapeutic target in triple negative breast cancer. Oncotarget 8:2971-2983
Mahajan, Kiran; Malla, Pavani; Lawrence, Harshani R et al. (2017) ACK1/TNK2 Regulates Histone H4 Tyr88-phosphorylation and AR Gene Expression in Castration-Resistant Prostate Cancer. Cancer Cell 31:790-803.e8