299 research outputs found
Abstract 1333: CD4 Th1 cytokines and HER-2/HER-3 blockade induces tumor apoptosis in breast cancer
No full textAbstract
Background: The therapeutic benefit of blocking HER2/HER3 signaling in breast cancer (BC) has been demonstrated by several studies. We have previously shown that the pleiotropic T-helper type 1 (Th1) cytokines IFN-γ and TNF-α induce senescence in BC cells and that all BC cell lines tested express IFN-γ and TNF-α receptors by western blot analysis. We have also demonstrated an inverse correlation between the HER2 expression level and the senescence induced by the treatment with both cytokines. Moreover, simultaneous HER-2/HER-3 blockade significantly enhanced cytokine-induced senescence. Here, we studied whether these Th1 cytokines induce apoptosis of HER-2 expressing BC cells and assessed the impact of IFN-γ and TNF-α with simultaneous HER-2 and HER-3 blockade on permanent tumor abrogation.
Results: To determine the Th1-mediated effects on HER2high (SK-BR-3, BT-474), HER2intermediate (MCF-7, T-47D), and HER2low (MDA-MB-231) human BC cell lines in vitro, we performed co-culture of increasing number of HER2 Class II peptide-specific CD4+ T-cells (generated by priming CD4+ T cells with HER2 peptide loaded type-1 polarized DCs) with BC cells using a transwell culture system. This resulted in a cell number-dependent apoptosis of SK-BR-3 and MCF-7, but not MDA-MB-231 cells compared with CD4+ T cells primed either with immature dendritic cells (DC) or mature DC plus irrelevant (Class II BRAF) peptide. In addition, SK-BR-3 cells incubated with supernatants from the CD4+ T cells-immature DC or mature DC co-culture demonstrated similar results. Compared with controls, HER2-specific Th1 cells generated a 25-fold increase in SK-BR-3 apoptosis by DAPI staining. Neutralizing antibodies against IFN-γ and TNF-α significantly reduced apoptosis induction. Also, IFN-γ and TNF-α treatment resulted in significant apoptosis of SK-BR-3 and MCF-7, but not MDA-MB-231 cells. However, MDA-MB-231 cells transfected with a wild type HER2 plasmid, were highly susceptible to cytokine-induced apoptosis. Treatment of HER-2-depleted cells (by RNAi) with IFN-γ and TNF-α resulted in an increased apoptotic phenotype. Although the combined treatment of IFN-γ and TNF-α in HER3-depleted cells did not enhanced the apoptosis, the double knock down with HER2 and HER3 RNAi strongly increased apoptosis induction as observed by western blot analysis of active caspase-3 and flow cytometric analysis of Anexin V-PI staining. Interestingly, the double depleted cells treated with IFN-γ alone evidenced slightly higher cleaved caspase-3 levels than TNF-α alone but the combined treatment with both cytokines had a strong synergistic effect.
Conclusions: Our results establish a role for IFN-γ and TNF-α in inducing tumor apoptosis in BC. An effective CD4 Th1 response, combined with HER-2 and HER-3 blockade can significantly drive tumor apoptosis that can be explored as treatment to effectively eliminate residual BC cells and prevent recurrence.
Citation Format: Cinthia Rosemblit, Jashodeep Datta, Erik Berk, Brian J. Czerniecki. CD4 Th1 cytokines and HER-2/HER-3 blockade induces tumor apoptosis in breast cancer. [abstract]. In: Proceedings of the 106th Annual Meeting of the American Association for Cancer Research; 2015 Apr 18-22; Philadelphia, PA. Philadelphia (PA): AACR; Cancer Res 2015;75(15 Suppl):Abstract nr 1333. doi:10.1158/1538-7445.AM2015-133
Oncodriver inhibition and CD4+ Th1 cytokines cooperate through Stat1 activation to induce tumor senescence and apoptosis in HER2+ and triple negative breast cancer: implications for combining immune and targeted therapies
No full textIn patients with HER2-expressing breast cancer many develop resistance to HER2 targeted therapies. We show that high and intermediate HER2-expressing cancer cell lines are driven toward apoptosis and tumor senescence when treated with either CD4+ Th1 cells, or Th1 cytokines TNF-α and IFN-γ, in a dose dependent manner. Depletion of HER2 activity by either siRNA or trastuzumab and pertuzumab, and subsequent treatment with either anti-HER2 Th1 cells or TNF-α and IFN-γ resulted in synergistic increased tumor senescence and apoptosis in cells both sensitive and cells resistant to trastuzumab which was inhibited by neutralizing anti-TNF-α and IFN-γ. Th1 cytokines induced minimal senescence or apoptosis in triple negative breast cancer cells (TNBC); however, inhibition of EGFR in combination with Th1 cytokines sensitized those cells causing both senescence and apoptosis. TNF-α and IFN-γ led to increased Stat1 phosphorylation through serine and tyrosine sites and a compensatory reduction in Stat3 activation. Single agent IFN-γ enhanced Stat1 phosphorylation on tyrosine 701 and similar effects were observed in combination with TNF-α and EGFR inhibition. These results demonstrate Th1 cytokines and antioncodriver blockade cooperate in causing tumor senescence and apoptosis in TNBC and HER2-expressing breast cancer, suggesting these combinations could be explored as non-cross-reactive therapy preventing recurrence in breast cancer.Fil: Rosemblit, Cinthia. H. Lee Moffitt Cancer Center; Estados Unidos. University of Pennsylvania; Estados Unidos. Consejo Nacional de Investigaciones Científicas y Técnicas; ArgentinaFil: Datta, Jashodeep. University of Pennsylvania; Estados UnidosFil: Lowenfeld, Lea. University of Pennsylvania; Estados UnidosFil: Xu, Shuwen. University of Pennsylvania; Estados UnidosFil: Basu, Amrita. H. Lee Moffitt Cancer Center; Estados UnidosFil: Kodumudi, Krithika. H. Lee Moffitt Cancer Center; Estados UnidosFil: Wiener, Doris. H. Lee Moffitt Cancer Center; Estados UnidosFil: Czerniecki, Brian J.. H. Lee Moffitt Cancer Center; Estados Unidos. University of Pennsylvania; Estados Unido
Identification of Patients for Adjuvant Therapy After Resection of Carcinoma of the Extrahepatic Bile Ducts: A Propensity Score-Matched Analysis
No full textBACKGROUND: Resectability rates for extrahepatic cholangiocarcinoma have increased over time, but long-term survival after resection alone with curative intent remains poor. Recent series suggest improved survival with adjuvant therapy. Patient subsets benefiting most from adjuvant therapy have not been clearly defined. METHODS: Patients with extrahepatic cholangiocarcinoma who underwent resection with curative intent and received adjuvant therapy (chemotherapy ± radiotherapy) or surgery alone (SA) were identified in the U.S. National Cancer Data Base (2004-2014). Cox regression identified covariates associated with overall survival (OS). Adjuvant therapy and SA cohorts were matched (1:1) by propensity scores based on the survival hazard in Cox modeling. Overall survival was compared by Kaplan-Meier estimates. RESULTS: Of 4872 patients, adjuvant chemotherapy was used frequently for 2416 (49.6%), often in conjunction with radiotherapy (RT) (n = 1555, 64.4%). Adjuvant chemotherapy with or without RT was used increasingly for cases with higher T classification [reference: T1-2; T3: 1.36; 95% confidence interval (CI), 1.19-1.55; T4: 1.77; 95% CI 1.38-2.26], nodal positivity [odds ratio (OR), 1.26; 95% CI 1.01-1.56], lymphovascular invasion (OR 1.21; 95% CI 1.01-1.46), or margin-positive resection (OR 1.85; 95% CI 1.61-2.12), and was associated with significant improvements in OS for each high-risk subset in the propensity score-matched cohort. Adjuvant therapy was associated with improved median OS for hilar tumors (40.0 vs 30.6 months; p = 0.025) but not distal tumors (33.0 vs 30.3 months; p = 0.123). Chemoradiotherapy was associated with superior outcomes compared with chemotherapy alone in the subset of margin-positive resection [hazard ratio (HR), 0.63; 95% CI 0.42-0.94]. CONCLUSIONS: Adjuvant multimodality therapy is associated with improved survival for patients with resected extrahepatic cholangiocarcinoma and high-risk features
Recommended from our members
Immunotherapy for Solid Tumors: A Comprehensive Roadmap for the Surgical Oncologist
No full textInternational students in United States’ medical schools: does the medical community know they exist?
No full textBackground: Matriculation of international students to United States’ (US) medical schools has not mirrored the remarkable influx of these students to other US institutions of higher education. Methods: While these students’ numbers are on the rise, the visibility for their unique issues remains largely ignored in the medical literature. Results: These students are disadvantaged in the medical school admissions process due to financial and immigration-related concerns, and academic standards for admittance also continue to be significantly higher compared with their US-citizen peers. Furthermore, it is simply beyond the mission of many medical schools – both public and private – to support international students’ education, especially since federal, state-allocated or institutional funds are limited and these institutions have a commitment to fulfill the healthcare education needs of qualified domestic candidates. In spite of these obstacles, a select group of international students do gain admission to US medical schools and, upon graduation, are credentialed equally as their US-citizen counterparts by the Accreditation Council for Graduate Medical Education (ACGME). However, owing to their foreign citizenship, these students have visa requirements for post-graduate training that may adversely impact their candidacy for residency placement. Conclusion: By raising such issues, this article aims to increase the awareness of considerations pertinent to this unique population of medical students. The argument is also made to support continued recruitment of international students to US medical schools in spite of these impediments. In our experience, these students are not only qualified to tackle the rigors of a US medical education, but also enrich the cultural diversity of the medical student body. Moreover, these graduates could effectively complement the efforts to augment US physician workforce diversity while contributing to healthcare disparity eradication, minority health issues, and service in medically underserved areas
Recommended from our members
Abstract C020: Neutrophil-mediated stromal-tumor IL-6/STAT-3 signaling underlies the association between neutrophil-to-lymphocyte ratio dynamics and chemotherapy response in localized pancreatic cancer: A hybrid clinical-preclinical study
No full textAbstract
Background: Partial/complete pathologic response following neoadjuvant chemotherapy (NAC) in pancreatic cancer (PDAC) patients undergoing pancreatectomy is associated with improved survival. We sought to determine whether neutrophil-to-lymphocyte ratio (NLR) dynamics predict pathologic response following chemotherapy in PDAC, and if manipulating NLR impacts chemosensitivity in preclinical models and uncovers potential mechanistic underpinnings underlying these effects. Methods: Pathologic response in PDAC patients (n=94) undergoing NAC and pancreatectomy (7/2015-12/2019) was dichotomized as partial/complete or poor/absent (case-cohort design). Bootstrap-validated multivariable models assessed associations between pre-chemotherapy NLR (%neutrophils÷%lymphocytes) or NLR dynamics during chemotherapy (ΔNLR=pre-surgery—pre-chemotherapy NLR) and pathologic response, disease-free survival (DFS), and overall survival (OS). To preclinically model effects of NLR attenuation on chemosensitivity, C57BL/6 mice (n=8-10/arm) were orthotopically injected with KrasG12D/+;Trp53fl/+;PdxCre(KPC) cells and randomized to vehicle, NLR-attenuating anti-Ly6G, gemcitabine/paclitaxel, or gemcitabine/paclitaxel+anti-Ly6G treatments. Results: In 94 PDAC patients undergoing NAC (median:4 months), pre-chemotherapy NLR (P<0.001) and ΔNLR attenuation during NAC (P=0.002) were independently associated with partial/complete pathologic response. An NLR score=pre-chemotherapy NLR+ΔNLR correlated with DFS (P=0.006) and OS (P=0.002). Upon preclinical modeling, combining NLR-attenuating anti-Ly6G treatment with gemcitabine/paclitaxel—compared with gemcitabine/paclitaxel or anti-Ly6G alone—not only significantly reduced tumor burden and metastatic outgrowth, but also augmented tumor-infiltrating CD107a+-degranulating CD8+ T-cells (P<0.01) while dampening inflammatory cancer-associated fibroblast (CAF) polarization (P=0.006) and chemoresistant IL-6/STAT-3 signaling in vivo. Neutrophil-derived IL-1β emerged as a novel mediator of stromal inflammation, inducing inflammatory CAF polarization and CAF-tumor cell IL-6/STAT-3 signaling in ex vivo co-cultures. Conclusions: Therapeutic strategies to mitigate neutrophil-CAF-tumor cell IL-1β/IL-6/STAT-3 signaling during NAC may improve pathologic responses and/or survival in PDAC.
Citation Format: Iago De Castro Silva, Anna Bianchi, Nilesh Deshpande, Prateek Sharma, Siddharth Mehra, Peter Hosein, Deukwoo Kwon, Nipun Merchant, Jashodeep Datta. Neutrophil-mediated stromal-tumor IL-6/STAT-3 signaling underlies the association between neutrophil-to-lymphocyte ratio dynamics and chemotherapy response in localized pancreatic cancer: A hybrid clinical-preclinical study [abstract]. In: Proceedings of the AACR Special Conference on Pancreatic Cancer; 2022 Sep 13-16; Boston, MA. Philadelphia (PA): AACR; Cancer Res 2022;82(22 Suppl):Abstract nr C020
Recommended from our members
Neoadjuvant FOLFIRINOX fails to extend survival in resectable pancreatic cancer
No full text[...]a prespecified subgroup analysis of resectable patients in the largest of these trials, PREOPANC, failed to demonstrate survival benefit. [...]dose reductions were observed in over twothirds of patients in the neoadjuvant arm - not typical of contemporary practice at experienced centers routinely delivering these therapies in the neoadjuvant setting. [...]the bizarre association between higher rates of margin-Znodenegative resection with worse survival in the neoadjuvant arm of this trial is difficult to reconcile with multiple lines of prior evidence
Abstract 4079: Reversal of immune evasion mediated by HER2 requires both humoral and cellular HER-2 targeted immune interventions
No full textAbstract
Objectives: HER2 over-expression in breast cancer is associated with a poor prognosis and HER2 is an intense target of immunotherapy with both humoral and T cell -base approaches. HER2 expression is also known to facilitate the escape of tumor cells from immune surveillance by down-regulating MHC class I, resulting in a reduced sensitivity to CTL lysis. Restoring tumor cell MHC Class I expression and reversing their immune evasion are therefore important strategies in the design of immunotherapy against HER2 expressing tumors. In this study we investigated the effects of monoclonal antibody trastuzumab and T help-1 associated cytokines IFN-γ and TNF-A on HLA Class I expression and the susceptibilities to CTL lysis of human HER2 expressing tumor cell lines.
Methods: Breast cancer cell lines MCF7, SKBR3, BT474 and ovarian cancer cell line SKOV3 were treated with trastuzumab only, combination of IFN-γ and TNF-A or trastuzumab with IFN-γ and TNF-A. Cell surface expressions of HLA ABC were examined by flow cytometric analysis. The susceptibility of tumor cells to HER2 specific CD8+ T cell recognition and CTL lysis were assessed by measurement of CD8+ T cell IFN-γ secretion and flow cytometric analysis respectively.
Results: Th1 cytokines IFN-γ and TNF-A dramatically increased HLA ABC expressions on all tumor lines compared to untreated cells, [ MCF7 (p=0.04); SKBR3 (p=0.02); SCOV3 (p=0.02) and BT474 (p=0.04)], while trastuzumab alone showed little impact on HLA ABC up-regulation. When treated with a combination of trastuzumab, IFN-γ and TNF-A tumor cells displayed synergistically enhanced HLA ABC expression [MCF7 (p=0.02); SKBR3 (p=0.004); SKOV3 (0.006) and BT474 (p=0.03)]. We observed that IFN-γ and TNF-A treatment was able to remarkably increase CD8+ T cell recognition and CTL lysis of low and intermediate HER2 expressing tumors [ MCF7(p=0.03) and SKOV3(0.02)], but not High HER2 tumor[ SKBR3(p=0.07)]. Only a combined treatment of trastuzumab, IFN-γ and TNF-A rendered SKBR3 susceptible to a significant killing by CTL (p=0.02). To investigate the impact of other HER family members EGFR and HER3 on MHC class I expression , HER2 over-expressing breast cancer cell lines BT474 and SKBR3 were pre-treated with EGF or Heregulin, then subjected to IFN-γ and TNF-A treatments. Activation of HER1 and HER3 signaling made HER2 over-expressing cells resistant to the effect of MHC class I up-regulation by IFN-γ and TNF-A.
Conclusion: Both humoral and cellular immune HER-2 targeted interventions are required to reverse immune escape mediated by HER2; in addition to targeting HER2, targeting HER1 and HER3 signaling elements should also be included in order to abrogate the cross talk between HER family members and more effectively inhibit the HER2 signaling pathway.
Citation Format: Shuwen Xu, Jessica Cintolo, Jashodeep Datta, Cinthia Rosemblit, Erik Berk, Julia Terhune, Elizabeth Fitzpatrick, Brian Czerniecki. Reversal of immune evasion mediated by HER2 requires both humoral and cellular HER-2 targeted immune interventions. [abstract]. In: Proceedings of the 105th Annual Meeting of the American Association for Cancer Research; 2014 Apr 5-9; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2014;74(19 Suppl):Abstract nr 4079. doi:10.1158/1538-7445.AM2014-407
Recommended from our members
Abstract PO-046: Dissecting the role of tumor-intrinsic Cxcl1 in mediating immune exclusion in Ras-p53 cooperative pancreatic cancer
No full textAbstract INTRODUCTION: Ras-p53 cooperative mutations, which define >50% of human pancreatic ductal adenocarcinoma (PDAC), promote an immune-excluded tumor microenvironment. In Ras-p53 murine models that phenocopy human PDAC, overexpression of the myeloid chemo-attractant Cxcl1 is associated with T-cell exclusion. We sought to dissect the role of tumor-intrinsic Cxcl1 in mediating immune exclusion in Ras-p53 PDAC. METHODS: Single-cell immune deconvolution and Cxcl1 expression were queried in TCGA PDAC samples (n=150). Single-cell RNA sequencing (scRNAseq) data in KrasG12D/+;Trp53fl/+;Pdx1Cre (KPC WT) genetically engineered mice (GEM) were interrogated. CRISPR/Cas9-based genome editing was used to silence Cxcl1 in KPC tumor cells (KPC-Cxcl1KO). Multiparameter flow cytometry-based immunophenotyping of tumor-infiltrating and circulating myeloid/T-cell populations was performed in in-vivo orthotopic models utilizing KPCWT / KPC-Cxcl1KO cells. RESULTS: In human TCGA samples, Cxcl1 overexpression was strongly associated with expression of its cognate receptor CXCR2, reduced CD4+/CD8+ T-cells, and increased CD33+ myeloid-derived suppressor cells (MDSCs). In KPC GEMs, scRNAseq revealed that CXCR2 expression was exclusively present on the polymorphonuclear MDSC subset (PMN-MDSC). Exogenous Cxcl1 conditioning in murine bone marrow-derived cells induced PMN-MDSC polarization and upregulated CXCR2. Genetic ablation of Cxcl1 in KPC tumor cells disrupted transwell migration of splenocyte-enriched MDSC in-vitro; this effect was dependent on CXCR2 expression on MDSCs. There was a dramatic reduction in tumor weights and metastatic outgrowth (p<0.0001), as well as circulating PMN-MDSC and CXCR2+-MDSC populations in KPC-Cxcl1KO compared with KPCWT mice (p=0.0006). Significantly fewer PMN-MDSC and M2-like macrophages infiltrated KPC-Cxcl1KO tumors, which was accompanied by dramatically higher CD4+/CD8+ T-cell infiltration (p<0.0001). While cytolytic effector CD8+ T-cell populations (CD44+62L-CD107a+) remained unchanged, non-cytolytic “exhausted” PD-1+CD8+ T-cells (PD-1+CD107a-) increased in KPC-Cxcl1KO vs. KPCWT tumors, providing rationale for addition of checkpoint blockade to reverse T-cell exhaustion. CONCLUSION: Tumor-intrinsic Cxcl1 appears a major contributor to immune exclusion in Ras-p53 cooperative PDAC. Targeting Cxcl1 represents a promising strategy to overcome MDSC-mediated immunosuppression and improve susceptibility to checkpoint blockade in PDAC. Citation Format: Anna Bianchi, Siddharth Mehra, Daisy Dai, Iago de Castro Silva, Prateek Sharma, Antonio Colaprico, Austin R. Dosch, Samara Singh, Nagaraj S. Nagathihalli, Nipun B. Merchant, Jashodeep Datta. Dissecting the role of tumor-intrinsic Cxcl1 in mediating immune exclusion in Ras-p53 cooperative pancreatic cancer [abstract]. In: Proceedings of the AACR Virtual Special Conference on Pancreatic Cancer; 2020 Sep 29-30. Philadelphia (PA): AACR; Cancer Res 2020;80(22 Suppl):Abstract nr PO-046
- …
