Abstract
Racial disparities in prostate cancer have not been well characterized on a genomic level. Here we show the results of a multi-institutional retrospective analysis of 1,152 patients (596 ...African-American men (AAM) and 556 European-American men (EAM)) who underwent radical prostatectomy. Comparative analyses between the race groups were conducted at the clinical, genomic, pathway, molecular subtype, and prognostic levels. The EAM group had increased
ERG
(
P
< 0.001) and
ETS
(
P
= 0.02) expression, decreased SPINK1 expression (
P
< 0.001), and basal-like (
P
< 0.001) molecular subtypes. After adjusting for confounders, the AAM group was associated with higher expression of
CRYBB2, GSTM3
, and inflammation genes (
IL33, IFNG, CCL4, CD3, ICOSLG
), and lower expression of mismatch repair genes (
MSH2
,
MSH6
) (p < 0.001 for all). At the pathway level, the AAM group had higher expression of genes sets related to the immune response, apoptosis, hypoxia, and reactive oxygen species. EAM group was associated with higher levels of fatty acid metabolism, DNA repair, and WNT/beta-catenin signaling. Based on cell lines data, AAM were predicted to have higher potential response to DNA damage. In conclusion, biological characteristics of prostate tumor were substantially different in AAM when compared to EAM.
Abstract
Background:
Certain solid malignancies like prostate cancer pose two major challenges for effective immunotherapy. The inherently low mutation load and spatial and temporal intra-tumor ...heterogeneity yields an immune exclusion and development of an “immune desert” within the tumor micro-environment (TME). Additionally, there is a response failure to immunomodulation, due to tumor/patient immunosuppressive mechanisms. In an effort to transform the prostate tumor environment into an immunogenic ecosystem, we are using PolyIC:LC as an immunemodulator. The novelty of this approach is a “host targeted”, in-situ “autovaccination” strategy, which uses the patient’s own tumor as the antigen source leading to activation of both an innate and adaptive immune response. As all patients will have their cancer removed after investigational therapy we can study baseline versus treatment induced changes in bio-specimens collected before, during, and after patients are exposed to PolyIC:LC. Correlative studies include characterization of tissue and systemic biomarkers of response using multiple platforms like cytometry by time of flight (CyTOF), RNA-seq, whole exome sequencing, seromics, TCR-sequencing, neoantigen specific T-cell responses, as well as assessing circulating tumor DNA (ctDNA) at multiple time points to determine the potential in detecting tumor response to immunemodulation.
Methods:
This is a Phase I dose escalation study (NCT03262103) seeking to determine a safe dose and schedule of intratumoral (IT) plus intramuscular (IM) PolyIC:LC injections prior to radical prostatectomy in patients with prostate cancer. The dose and frequency of IT PolyIC:LC will be increased in successive cohorts using a 3+3 design and traditional dose escalation rules. The dose and schedule of IM PolyIC:LC will remain fixed in successive cohorts. The study will consist of 24 enrolled subjects, recruited into cohorts consisting of a minimum of 3 and maximum of 6 patients per cohort. The first cohort, consisting of three patients, has been completed. Recruitment for the second cohort has begun with two patients already enrolled. The inclusion criteria extend to patients diagnosed with high risk (Gleason 7-10, cT2a-cT3b) clinically localized prostate cancer with no prior hormonal or radiation therapy and with plans to undergo radical prostatectomy. Week 1 serves as the priming course with the patient coming in for a pre-treatment biopsy followed by an IT injection. Weeks 3-6 consist of a booster treatment course with IM injections two times a week. Weeks 7-9 are a rest period with no injections followed by radical prostatectomy at week 10. Blood is drawn at weeks 1, 3, 6, and 9. At the time of surgery, blood, tissue, and lymph node are collected for research purposes. Following each IT and IM injection, the subject remains in clinic for monitoring for at least 1 hour or 30 minutes respectively. Patients are seen 6 weeks post-prostatectomy as per standard of care. The next follow-up visit is approximately 3 months following surgery where the first post-operative PSA check is performed. Assuming PSA levels are undetectable, the patient is followed up at routine intervals for PSA testing.
Citation Format: Sujit S. Nair, Rachel Weil, Elena Gonzalez-Gugel, Marcia Meseck, Alex Rubinsteyn, Yulia Kodysh, Akriti Gupta, Keerthi Sadanala, Kacie Schlussel, Kamala Bhatt, Avinash Reddy, Rajan Patel, Tin Thawte, Adam Farkas, Siarhei Dzedzik, Kenneth Haines, Julia Wagner, Macy Robison, Cynthia Knauer, Andres Salazar, Matthew Galsky, Nina Bhardwaj, Ashutosh Tewari. Phase I study of in situ autologous vaccination for prostate cancer in a neo-adjuvant setting abstract. In: Proceedings of the American Association for Cancer Research Annual Meeting 2019; 2019 Mar 29-Apr 3; Atlanta, GA. Philadelphia (PA): AACR; Cancer Res 2019;79(13 Suppl):Abstract nr CT096.
In last couple of decades the use of natural compounds like flavonoids as chemopreventive agents has gained much attention. Our current study focuses on identifying chemopreventive flavonoids and ...their mechanism of action on human prostate cancer cells. Human prostate cancer cells (PC3), stably transfected with activator protein 1 (AP-1) luciferase reporter gene were treated with four main classes of flavonoids namely flavonols, flavones, flavonones, and isoflavones. The maximum AP-1 luciferase induction of about 3 fold over control was observed with $20\;{\mu}M$ concentrations of quercetin, chrysin and genistein and $50\;{\mu}M$ concentration of kaempferol. At higher concentrations, most of the flavonoids demonstrated inhibition of AP-1 activity. The MTS assay for cell viability at 24 h showed that even at a very high concentration $(500\;{\mu}M)$, cell death was minimal for most of the flavonoids. To determine the role of MAPK pathway in the induction of AP-1 by flavonoids, Western blot of phospho MAPK proteins was performed. Four out of the eight flavonoids namely kaempferol, apigenin, genistein and naringenin were used for the Western Blot analysis. Induction of phospho-JNK and phospho-ERK activity was observed after two hour incubation of PC3-AP1 cells with flavonoids. However no induction of phospho-p38 activity was observed. Furthermore, pretreating the cells with specific inhibitors of JNK reduced the AP-1 luciferase activity that was induced by genistein while pretreatment with MEK inhibitor reduced the AP-1 luciferase activity induced by kaempferol. The pharmacological inhibitors did not affect the AP-1 luciferase activity induced by apigenin and naringenin. These results suggest the possible involvement of JNK pathway in genistein induced AP-1 activity while the ERK pathway seems to play an important role in kaempferol induced AP-1 activity.
Abstract PURPOSE To test the safety of sequential intratumoral plus systemic intramuscular injection of poly-ICLC, and its efficacy in priming antitumor immune responses in patients with prostate ...cancer (PCa). EXPERIMENTAL PROCEDURES This is an open-label dose-escalating phase 1 neoadjuvant clinical trial of poly-ICLC (NCT03262103) in 12 patients diagnosed with clinically localized PCa with plans to undergo radical prostatectomy (RP). Poly-ICLC was administered intratumorally (Artemis MRI-TRUS-guided) and intramuscularly (e.g., deltoid muscle). Comprehensive transcriptional profiling of tissues, phenotypic and transcriptional analysis of longitudinally collected peripheral blood, and analysis of the prostate tumor microenvironment (TME) were performed before and after poly-ICLC treatment in order to identify innate and adaptive antitumor immune responses within the tumor and in peripheral blood. This trial is the first to use human intratumoral immunotherapy instead of systemic immunomodulation for high-risk PCa patients. RESULTS Poly-ICLC was well tolerated (safe) in all 12 patients. There were no dose-limiting toxicities or TEAE (treatment-emergent adverse events) withdrawals during treatment. Eight of 10 evaluable patients (80%) had undetectable PSA (<0.1 ng/ml) at 1 year of follow-up post-RP. Eight of 12 evaluable patients (66.7%) and 7 of 10 patients (70%) with biopsy Gleason 8-10 had a lower Gleason score in the post-treatment RP specimen. Transcriptome profiling of paired biopsy and RP specimens showed significant upregulation of gene signatures associated with immune cell infiltration and TP53-associated metabolic genes, but downregulation of gene signatures associated with metastasis and DNA replication. Genes upregulated by poly-ICLC were associated with a good prognosis. Treatment with poly-ICLC increased systemic immune responses, as demonstrated by an increase in cytolytic signatures, T- and NK-cell signatures, and NK-cell subsets in the blood. Multiplex immunohistochemistry analysis revealed a significant increase in the densities of CD4- and CD8-T cells and B cells, as well as evidence of tertiary lymphoid structures (TLS) within the TME of post-treatment RP specimens compared to baseline. CONCLUSIONS Poly-ICLC treatment can reliably transform the cold TME of PCa into a hot, immune-enhanced ecosystem. The identified baseline and response biomarkers, tertiary lymphoid structures, potential clinical benefit, and immunologic correlates require validation in larger studies. Citation Format: Sujit S. Nair, Dimple Chakravarty, Sreekumar Balan, Alexander Hakansson, Manuel Duval, Elai Davicioni, Yang Liu, Swati Bhardwaj, Tin Htwe Thin, Monica Garcia-barros, Kenneth Haines, Majd Al Shaarani, Rachel Weil, Marcia Meseck, Parita Ratnani, Monali Fatterpekar, Ivan Jambor, Elena Gonzalez-Gugel, Adam Farkas, Vinayak Wagaskar, Kacie Schlussel, Cristina Pasat-karasik, Kamala Bhatt, Zachary Dovey, Adriana Pedraza, Akriti Gupta, Dara Lundon, Ante Peros, Sneha Parekh, Lily Davenport, Xiangfu Zhang, Raghav Gupta, Macy Robison, Cynthia Knauer, Ethan Ellis, Dmitry Rykunov, Boris Reva, Babu Padanilam, Matthew D. Galsky, Rachel Brody, Mani Menon, Andres Salazar, Nina Bhardwaj, Ashutosh K. Tewari. Prostate cancer in situ autovaccination with the intratumoral viral mimic poly-ICLC: Making a cold tumor hot abstract. In: Proceedings of the American Association for Cancer Research Annual Meeting 2024; Part 2 (Late-Breaking, Clinical Trial, and Invited Abstracts); 2024 Apr 5-10; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2024;84(7_Suppl):Abstract nr CT023.
Abstract
Estrogen-induced breast carcinogenesis is characterized by global changes in histone modifications. KDM1 (also known as LSD1) a histone demethylase enzyme, plays a key role in establishing ...specific histone methylation marks at ERα target gene promoters. Emerging evidence suggests histone methylation imposes ligand dependency; and deregulation of this epigenetic pathway potentially contributes to hormonal independence and adaptive resistance in breast cancer. Our study examined the therapeutic efficacy of pargyline, a KDM1 inhibiting drug, and evaluated the therapeutic benefit to curb growth of therapy resistant breast cancer cells. We used several model cell lines shown to exhibit resistance to hormonal therapy including MCF7-HER2, MCF7-Tam, MCF7-Ca-LTLT, MCF7-PELP1 and parental MCF7 cells as controls. Reporter gene assays showed KDM1 enhanced ERα- mediated transcription in therapy resistant cells. KDM1 functionally interacts with ERα coregulator PELP1 and is co-recruited with PELP1 to ERα target genes. Pargyline treatment substantially inhibited ERα transactivation functions. ChIP analysis revealed distinct activating histone methylation modifications at growth regulatory ERα target genes in aggressively growing and therapy resistant breast cancer cells. Breast cancer models treated with pargyline facilitated reversal of the observed specific modifications and thereby inhibited the growth of breast cancer cells in vitro and in vivo nude mice models. Pargyline also showed significant effect in blocking local estrogen synthesis via alterations in histone methylation modifications at the aromatase promoter. Combinational therapy using three agents that: (a) block ERα's nuclear actions (tamoxifen or letrozole), (b) ERα's extranuclear actions (dasatinib) and (c) ERα's epigenetic modifications (pargyline) showed most promising therapeutic effect compared to single agent therapy on the growth of therapy resistant cells. Our results suggest that histone methylation modifications play a role in therapy resistance and validate the therapeutic potential of pargyline in combinational therapies. Collectively, our results suggests targeting KDM1 axis with pargyline in combination with current endocrine therapies will have better therapeutic effect and may inhibit or delay development of hormonal resistance. This study is funded by Komen grant KG090447.
Citation Format: {Authors}. {Abstract title} abstract. In: Proceedings of the 101st Annual Meeting of the American Association for Cancer Research; 2010 Apr 17-21; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2010;70(8 Suppl):Abstract nr 4613.
Estrogen receptor (ER) signaling plays an important role in breast cancer progression and ER functions are influenced by coregulatory proteins. Proline-, glutamic acid- and leucine-rich protein 1 ...(PELP1) is a nuclear receptor coregulator that plays an important role in ER signaling. Its expression is deregulated in hormonal cancers. We identified PELP1 is a novel cyclin dependent kinase (CDK) substrate. Using site-directed mutagenesis, and
in vitro
kinase assays, we identified Ser477 and Ser991 of PELP1 as CDK phosphorylation sites. Using the PELP1 S991-phospho-specific antibody, we show that PELP1 is hyper-phosphorylated during cell cycle progression. Model cells stably expressing the PELP1 mutant that lack CDK sites had defects in E2-mediated cell cycle progression and significantly affected PELP1-mediated oncogenic functions
in vivo
. Mechanistic studies showed that PELP1 modulates transcription factor E2F1 transactivation functions, that PELP1 is recruited to pRb/E2F target genes, and that PELP1 facilitates ER signaling cross talk with cell cycle machinery. We conclude that PELP1 is a novel substrate of interphase CDKs, and that its phosphorylation is important for the proper function of PELP1 in modulating hormone-driven cell cycle progression and also for optimal E2F transactivation function. Because the expression of both PELP1 and CDKs are deregulated in breast tumors, CDK-PELP1 interactions will have implications in breast cancer progression.
Histone methylation has a key role in oestrogen receptor (ERalpha)-mediated transactivation of genes. Proline glutamic acid and leucine-rich protein 1 (PELP1) is a new proto-oncogene that functions ...as an ERalpha co-regulator. In this study, we identified histone lysine demethylase, KDM1, as a new PELP1-interacting protein. These proteins, PELP1 and KDM1, were both recruited to ERalpha target genes, and PELP1 depletion affected the dimethyl histone modifications at ERalpha target genes. Dimethyl-modified histones H3K4 and H3K9 are recognized by PELP1, and PELP1 alters the substrate specificity of KDM1 from H3K4 to H3K9. Effective demethylation of dimethyl H3K9 by KDM1 requires a KDM1-ERalpha-PELP1 functional complex. These results suggest that PELP1 is a reader of H3 methylation marks and has a crucial role in modulating the histone code at the ERalpha target genes. PUBLICATION ABSTRACT
Based on the recently established role for the master coregulator MTA1 and MTA1-containing nuclear remodeling complexes, in oncogenesis and inflammation, we explored the links between parasitism by ...the carcinogenic liver fluke
Opisthorchis viverrini
and this coregulator using both an
Mta1−/−
mouse model of infection and a tissue microarray of liver fluke induced human cholangiocarcinomas. Intense foci of inflammation and periductal fibrosis in the liver and kidney of wild type,
Mta1+/+
mice were evident at 23 days post-infection with
O. viverrini
. In contrast, little inflammatory response was observed in the same organs of infected
Mta1−/−
mice. Livers of infected
Mta1+/+
mice revealed strong upregulation of fibrosis-associated markers such as cytokeratins 18, 19, and annexin-2, as determined by both by immunostaining and by reverse transcription PCR compared to infected
Mta1−/−
mice. CD4 expression was upregulated by infection in the liver of both experimental groups; however, its levels were several folds higher in the
Mta1
+/+ mice than in infected
Mta1−/−
mice.
Mta1−/−
infected mice also exhibited significantly higher systemic and hepatic levels of host cytokines such as IL-12p70, IL-10 and IFN-γ compared to the levels of these cytokines in the
Mta1+/+
mice, suggesting an essential role of MTA1 in the cross-regulation of the Th1 and Th2 responses, presumably due to chromatin remodeling of the target chromatin genes. Immunohistochemical analysis of ~300 liver tissue cores from confirmed cases of
O. viverrini
induced cholangiocarcinoma showed that MTA1 expression was elevated in more than 80% of the specimens. These findings suggest that MTA1 status plays an important role in conferring an optimal cytokine response in mice following infection with
O. viverrini
and is a major player in parasite-induced cholangiocarcinoma in humans.
Schistosoma haematobium
is responsible for two-thirds of the world’s 200-400 million cases of human schistosomiasis. It is a group 1 carcinogen and a leading cause of bladder cancer which occurs ...following years of chronic inflammation, fibrosis and hyper-proliferation in the host liver. The co-evolution of blood flukes of the genus
Schistosoma
and their human hosts is paradigmatic of long term parasite development, survival and maintenance in mammals. However, the contribution of host genes, especially those discrete from those of the immune system, necessary for parasite establishment and development remains poorly understood. The present study investigated the role of Metastasis-associated protein-1 gene (
Mta1)
product in the survival of
S. haematobium
. Significantly fewer
S. haematobium
worms and eggs were recovered from
Mta1
−/− than wild type mice. Comparative analysis of cytokine profiles indicated a loss of cytokine interdependence and aberrant Th1/Th2 cytokine response in the
Mta1
−/− mice compared to age-matched wild type mice. By utilizing this
Mta1
-null mouse model, we identified a distinct, contribution of the mammalian MTA1 in establishing a productive host-parasite interaction and thus revealed a host factor critical for the optimal survival of schistosomes and successful parasitism. Moreover, MTA1 appears to play a significant role in driving inflammatory responses to schistosome egg induced hepatic granulomata reactions, and thus offers a survival cue for parasitism as well as an obligatory contribution of liver in schistosomiasis. These findings raise the possibility to develop intervention strategies targeting MTA1 to reduce the global burden of schistosomiasis, inflammation and neoplasia.
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