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Affinity Purification of EGFR, JUN, JNK, and FOS

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Objective To explore the efficacy of primary chemoradiation with cisplatin versus cetuximab with respect to HPV/p16 and smoking statuses. Methods We retrospectively reviewed patients from our center with locally advanced non-nasopharyngeal head and neck squamous cell carcinoma (HNSCC) who received primary chemoradiation with cisplatin or cetuximab between 2006 and 2018. Results The median OS for cisplatin (n = 66) was not reached versus 132 months when treated with cetuximab (n = 55) (p = 0.03). For HPV/p16-positive patients, we found the median OS for cisplatin (n = 34) was not reached versus 60 months with cetuximab (n = 21) (p = 0.036). In the smoking group, the median OS was not reached in the cisplatin group (n = 44) versus 60 months when treated with cetuximab (n = 32) (p = 0.03). Conclusion HPV/p16-positive and smoking cohorts treated with cisplatin-based chemoradiotherapy had a significantly better OS versus cetuximab.

Purpose: Interpersonal trust is linked to therapeutic factors of patient care, including adherence to treatment, continuity with a provider, perceived effectiveness of care, and clinical outcomes. Differences in interpersonal trust across groups may contribute to health disparities. We explored whether differences in interpersonal trust varied across three racial/ethnic groups. Additionally, we explored how different health care factors were associated with differences in trust. Methods: We conducted a cross-sectional, computer-administered survey with 600 racially and ethnically diverse adults in Chicago, IL, from a wide variety of neighborhoods. We used staged ordinal logistic regression models to analyze the association between interpersonal trust and variables of interest. Results: Interpersonal trust did not differ by racial or ethnic group. However, individuals with 0–2 annual doctor visits, those reporting having a “hard time” getting health care services, those answering “yes” to “Did you not follow advice or treatment plan because it cost too much?,” and those reporting waiting more than 6 days/never getting an appointment had significantly increased odds of low trust. We did not find differences across racial/ethnic groups. Conclusion: Our study suggests that access to health care and interactions within the health care setting negatively impact individual's trust in their physician.

Simple Summary Ovarian cancer has a dismal prognosis and innovative treatment options are necessary to improve survival. Because the microRNA let-7 is often lost in this and other cancers, and its loss is associated with poor prognosis, we focused on therapeutic strategies to replace it. We report that let-7 overexpression in patient-derived cells resulted in a loss of aggressiveness: inhibition of migration and invasion (associated with metastasis), repression of cancer stem cell attributes (necessary for tumor maintenance and recurrence), and promotion of cell death (required for sensitivity to chemotherapy drugs). Further, cells in which let-7 is overexpressed were more sensitive to PARP inhibitors, even in patients who otherwise could not benefit from these drugs. We show that let-7 reduces the expression of several genes that may contribute to these effects. These actions of let-7 add to the rationale for use of this miRNA as a treatment for selected ovarian cancer patients. Abstract High-grade serous carcinoma of the ovary is a deadly gynecological cancer with poor long-term survival. Dysregulation of microRNAs has been shown to contribute to the formation of cancer stem cells (CSCs), an important part of oncogenesis and tumor progression. The let-7 family of microRNAs has previously been shown to regulate stemness and has tumor suppressive actions in a variety of cancers, including ovarian. Here, we demonstrate tumor suppressor actions of let-7i: repression of cancer cell stemness, inhibition of migration and invasion, and promotion of apoptosis, features important for cancer progression, relapse, and metastasis. Let-7i over-expression results in increased sensitivity to the PARP inhibitor olaparib in samples without BRCA mutations, consistent with induction of BRCAness phenotype. We also show that let-7i inhibits the expression of several factors involved in the homologous recombination repair (HRR) pathway, providing potential mechanisms by which the BRCAness phenotype could be induced. These actions of let-7i add to the rationale for use of this miRNA as a treatment for ovarian cancer patients, including those without mutations in the HRR pathway.

Pineoblastoma is a rare, primitive, and malignant tumor arising from the parenchyma of the pineal gland. It typically metastasizes along the cerebral neural axis, with rare extraneural metastasis and even more rare intraosseous extraneural metastasis. A patient with pineoblastoma, initially treated with chemotherapy, presented 10 years after initial diagnosis with multiple osseous metastases including his pelvis, femur, and vertebrae, and is currently undergoing chemotherapy.

Background: Breast cancer (BC) is the leading cause of cancer death in Hispanic/Latina women nationwide. Hispanic women are more likely to be presented with advanced disease and might have adverse prognosis. Further, the Hispanics of Mexican-American origin might reflect different clinico-pathological characteristics as opposed to other Hispanics and ethnic groups. No previous largest studies comprised with Hispanics of Mexican-American origin explored tumor characteristics and compared to other ethnic groups. Thus, the aim of this study was to describe the clinico- pathological characteristics and disparities in breast cancer in this minority group at two tertiary care University- based medical centers in 2 states with a large Hispanic presence. Methods: After IRB approval, cancer registry was used to analyze the variables of 3,441 patients with breast cancer diagnosed and treated consecutively at two large tertiary University based medical centers in El Paso, TX and Loma Linda, CA between 2005-2015. Unadjusted and adjusted associations of race/ethnicity with cancer stage, hormone receptor status and treatment option were investigated, as well as comparison to other ethnic groups. Results: Overall 45.5% of the patients were Hispanic (n= 1566). Hispanics were more likely to be diagnosed at a younger age (57 years) compared to in non-Hispanic (NH) whites, more likely to have invasive ductal carcinoma type (82.7%) & triple negative disease (17.1%, 95%CI: 15% to 19%). 58.8% of Hispanics (95%CI: 56% to 61%) have HR+ & HER2- as opposed to 71% in NH whites. In addition, Hispanic individuals presented with advanced stages (III and IV) of BC (25.3%, 95% CI: 23% to 28%) similar to African Americans (25.4%), and had a lower proportion of lumpectomy versus mastectomy compared to NH whites (50%) but similar to African Americans (50%). Hispanic patients had the highest prevalence of triple negative BC (17.1% in Hispanics Versus 13.9 % in African Americans, versus 8.5% in NH whites). Hispanics also had significantly higher relative risk of HER2+/HR - disease (RRR=1.77, p Conclusions: This large multi-institutional study shows that Hispanics are diagnosed with breast cancer at a younger age, have a higher prevalence of triple negative and HER2 positive/HR- breast cancer, are diagnosed at more advanced stages of disease and undergo less lumpectomies compared to NH whites. Increased efforts geared toward early detection, improving awareness and access to health care is desperately needed in this rapidly increasing minority in the U.S. Citation Format: Otoukesh S, Nahleh Z, Mirshahidi HR, Nguyen AL, Botrus G, Badri N, Diab N, Alvarado A, Sanchez LA, Dwivedi A. “Disparities in breast cancer: A multi-institutional comparative analysis focusing on American Hispanics” [abstract]. In: Proceedings of the 2017 San Antonio Breast Cancer Symposium; 2017 Dec 5-9; San Antonio, TX. Philadelphia (PA): AACR; Cancer Res 2018;78(4 Suppl):Abstract nr P6-10-04.

We studied ovarian cancer patient‐derived cells to determine their epithelial vs. mesenchymal phenotype, and their stemness, migration, invasion, and tumor growth characteristics. Surprisingly, stemness could be dissociated from invasiveness. We observed that lower let‐7 levels are associated with the epithelial state and stemness, reliably predict self‐renewal and tumor burden in mice, and could contribute to prognosis calculations., Patient‐derived samples present an advantage over current cell line models of high‐grade serous ovarian cancer (HGSOC) that are not always reliable and phenotypically faithful models of in vivo HGSOC. To improve upon cell line models of HGSOC, we set out to characterize a panel of patient‐derived cells and determine their epithelial and mesenchymal characteristics. We analyzed RNA and protein expression levels in patient‐derived xenograft (PDX) models of HGSOC, and functionally characterized these models using flow cytometry, wound healing assays, invasion assays, and spheroid cultures. Besides in vitro work, we also evaluated the growth characteristics of PDX in vivo (orthotopic PDX). We found that all samples had hybrid characteristics, covering a spectrum from an epithelial‐to‐mesenchymal state. Samples with a stronger epithelial phenotype were more active in self‐renewal assays and more tumorigenic in orthotopic xenograft models as compared to samples with a stronger mesenchymal phenotype, which were more migratory and invasive. Additionally, we observed an inverse association between microRNA let‐7 (lethal‐7) expression and stemness, consistent with the loss of let‐7 being an important component of the cancer stem cell phenotype. We observed that lower let‐7 levels were associated with the epithelial state and a lower epithelial mesenchymal transition (EMT) score, more efficient spheroid and tumor formation, and increased sensitivity to platinum‐based chemotherapy. Surprisingly, in these HGSOC cells, stemness could be dissociated from invasiveness: Cells with lower let‐7 levels were more tumorigenic, but less migratory, and with a lower EMT score, than those with higher let‐7 levels. We conclude that let‐7 expression and epithelial/mesenchymal state are valuable predictors of HGSOC proliferation, in vitro self‐renewal, and tumor burden in vivo.

Introduction: In order to assess hematopoietic precursor cells, bone marrow biopsies are routinely conducted, which is painful for the patient and is not applicable for frequent testing. However, precursor, maturating, and matured cells are known to produce exosomes, which are released into peripheral blood. By analyzing these exosomes in plasma, bone marrow condition may be assessed noninvasively. Myeloid precursor cells express different surface markers than when they have matured. It is reasonable to speculate that these markers would be incorporated into exosomes. In this study, we developed a novel sandwich enzyme-linked immunosorbent assay (ELISA) by using a combination of monoclonal antibodies against 2 different leukocyte antigens, to account for the number of exosomes in plasma. Methods In 6 patients undergoing chemotherapy, venous blood was drawn at regular intervals before, during, and after respective chemotherapies was given. Various clones of monoclonal antibodies against CD11b, CD33, CD34, CD35, CD81, and CD117 were obtained, and antibodies were biotinylated by EZ link Sulfo-NHS-LC-Biotin. Anti-brain antibodies were switched with anti-leukocyte antibodies, and exosome activity was trended. Results CD11b+ and CD 117+ plates showed small changes during chemotherapy (within +/- 50% of the initial value), while plasma WBC count showed a dramatic decrease during chemotherapy. In the recovery phase of WBC after chemotherapy, CD11b+ and CD16+showed a greater than 2-fold increase, with the up-trend taking place earlier than its plasma WBC counterparts. CD 117+ CD33+ also showed an up-trend, but was slower than their CD11b+ and CD16+ counterpart. Finally, each exosome value was divided by other markers to calculate the ratio, and compared between pre and post chemotherapy blood. In half of the cases, there was not a difference, versus an almost double premature/mature ratio in post-chemotherapy blood in the other half of patients. Conclusion From data collected from pre-chemotherapy studies, the lack of correlation between ELISA data compared to plasma WBC data, indicates that plasma exosome analysis might not be suitable for the monitoring of acute bone marrow suppression. Additionally, in the recovery phase, surface markers correlation with WBC was present in only some of the markers, indicating that the assay sensitivity is dependent on the marker combinations. Overall, both myeloid precursor- and mature granulocyte-derived exosomes were increased during the recovery from chemotherapy in 3 out of 5 patients, and exosome profiles were different after chemotherapy in 2 out of 5 patients. Disclosures Mitsuhash: NanoSomiX:Current Employment.

Objective: Patient-derived samples present an advantage over current cell line models of high grade serous ovarian cancer (HGSOC), which are flawed in terms of being reliable and phenotypically faithful models of in vivo HGSOC. To improve upon cell line models of HGSOC, we characterized a panel of patient-derived cells to determine their epithelial and mesenchymal characteristics, invasiveness, proliferation, stemness, and in vivo growth. Experimental procedures: Patient-derived xenograft (PDX) models of HGSOC were analyzed in vitro for phenotypic (RNA, let-7 miRNA, and protein expression, flow cytometry) and functional aspects including growth in spheroids, wound healing assays, invasion assays and in vivo (orthotopic PDX) growth characteristics. Results: Samples fell along the spectrum from epithelial to mesenchymal, and all had hybrid characteristics. Those toward the more epithelial end of the spectrum were most active in self-renewal assays, and grew most robustly in orthotopic xenograft models. Chemoresistance correlated both with the mesenchymal state and with BRCA2 wild type status. Loss of microRNA let-7 (lethal-7) is an important component of the cancer stem cell phenotype, and we observed an inverse association between let-7 expression and the epithelial state. We observed lower levels of let-7, more efficient spheroid and tumor formation, and increased sensitivity to platinum-based chemotherapy in cells with the most epithelial phenotype. Conclusions: Surprisingly, in these HGSOC cells, stemness could be dissociated from invasiveness: epithelial cells (those with least let-7 expression) were less migratory, but more tumorigenic, than the mesenchymal cells with higher let-7 expression. We conclude that epithelial/mesenchymal state and let-7 expression are valuable predictors of HGSOC proliferation, in vitro self-renewal, and tumor burden in vivo. Citation Format: Juli Unternaehrer, Evgeny Chirshev, Nozomi Hojo, Antonella Bertucci, Linda Sanderman, Anthony Nguyen, Hanmin Wang, Tise Suzuki, Emmanuel Brito, Shannalee Martine, Christine Castañón, Saied Mirshahidi, Marcelo Vazquez, Kerby C. Oberg, Yevgeniya J. Ioffe. Epithelial-mesenchymal heterogeneity of high-grade serous ovarian carcinoma samples correlates with let-7 levels and predicts tumor growth and metastasis [abstract]. In: Proceedings of the Annual Meeting of the American Association for Cancer Research 2020; 2020 Apr 27-28 and Jun 22-24. Philadelphia (PA): AACR; Cancer Res 2020;80(16 Suppl):Abstract nr 1682.

Breast cancer (BC) is the leading cause of cancer death in Hispanic/Latino women nationwide. Hispanic women are more likely to be presented with advanced disease and adverse prognosis subtypes. The aim of this study is to describe the clinico‐ pathological characteristics and disparities in breast cancer in this group at two tertiary care University‐based medical centers. After IRB approval, Cancer registry was used to analyze the variables of 3441 patients with breast cancer diagnosed and treated consecutively at two large tertiary University based medical and cancer center database centers in El Paso, TX and Loma Linda, CA between 2005 and 2015. Association between race/ethnicity and cancer type, stage, hormone receptor status and treatment option were investigated. Overall 45.5% of the patients were Hispanic (n: 1566) and those were more likely to be diagnosed at a younger age (57 years) similar to African Americans, more likely to have invasive ductal carcinoma type (82.7%) & triple negative disease (17.1%, 95%CI: 15% to 19%). 58.8% of Hispanics (95%CI: 56% to 61%) have hormone receptor (HR)+ & HER2− as opposed to 71% in non‐Hispanic White people. In addition, Hispanic individuals presented with advanced stages of BC (25.3%, 95% CI: 23% to 28%) similar to African American (25.4%), and had a lower proportion of lumpectomy (50%) similar to African American (50%). When compared to African American patients, Hispanic patients had a higher prevalence of triple negative BC (17.11% in Hispanics Versus 13.86% in African American). Conclusion: Hispanics had significantly higher relative risk of advanced stages at presentation (Relative Risk Ratio (RRR) = 2.05, P

Objective Our objective was to explore whether there are differences in institutional trust across racial/ethnic groups and what factors might contribute to these differences. Methods We studied a convenience sample of 569 adults in Chicago grocery stores who self-identified as African American, Mexican-Hispanic, or white. We measured institutional trust and dichotomized responses into “high” and “low” trust. We used chi squared tests to examine differences in institutional trust across racial/ethnic groups and stepwise multivariable logistic regression to investigate how sociodemographic factors, health care access, health care usage, and previous negative experience with the health care system modified this relationship. Results In unadjusted analysis, race/ethnicity was significantly associated with institutional trust (p

INTRODUCTION: Epithelial ovarian cancer (EOC) is the fifth deadliest malignancy in women in the United States, with a high death-to-incidence ratio and five-year survival of 30%. Aggressiveness and recurrence of EOC has been attributed to cancer stem cells (CSC) within tumors, which contribute to chemoresistance and relapse. Deregulation of miRNAs has been linked to cancer initiation, progression, and the stem cell state. We focus on let-7 miRNA, which is repressed in many cancers and is associated with decreased survival. Let-7 is essential for cell differentiation, and its repression is required for reprogramming. Decreased let-7 level in cancer is associated with stemness. OBJECTIVE: Purpose for our research is to understand let-7 expression and function in EOC cell-lines and patients-derived samples in order to understand mechanisms of its abnormal regulation and develop novel treatments. MATERIALS AND METHODS: Patient-derived samples are used along with EOC cell-lines. Over-expression of Let-7 is achieved by mimic transfection and confirmed via RT-qPCR. Level of miRNA and pluripotency markers mRNA is detected via RT-qPCR, and protein expression confirmed by Western blot. Functional assays to analyze cancer phenotype include wound healing, invasion and spheroid formation assays. Embryoid body (EB) formation is used to mimic murine early embryonic development and guided toward primitive streak via cytokines. Let-7 expression is analyzed via RT-qPCR. RESULTS: We demonstrate that patient-derived EOC samples have decreased Let-7 levels and increased expression of pluripotency markers, therefore, we hypothesize that up-regulation of let-7 in EOC will decrease aggressiveness and increase chemosensitivity. We have demonstrated that up-regulation of let-7 via mimic transfection reduced cancer stem cell properties demonstrated by reduction of pluripotency markers and spheroid formation. Let-7 also decreased in vitro migration and invasion. Our data show that let-7 suppresses stem cell-like phenotype of EOC, indicating that it may be a treatment option in conjunction with conventional chemotherapies. In order to understand deregulation of let-7 in cancer, understanding its normal regulation and function during development is essential. By using mESCs we demonstrate that let-7 is dynamically expressed, contradicting conventional belief that let-7 levels slowly increase upon differentiation. Previously published data demonstrating that Hmga2, a let-7 target, peaks and is required upon exit from pluripotency supports our results. CONCLUSION: In conclusion, let-7 is repressed in EOC, and its over-expression demonstrates tumor suppressive functions via decreasing stemness, migration, invasion, and spheroid formation. During development let-7 is dynamically expressed, demonstrating complex regulation. Citation Format: Evgeny A Chirshev, Anthony Nguyen, Hojo Nozomi, Hill Alyse, Juli Unternaehrer. MICRO-RNA LET-7 REGULATION AND FUNCTION IN OVARIAN CANCER AND EARLY EMBRYONIC DEVELOPMENT [abstract]. In: Proceedings of the 12th Biennial Ovarian Cancer Research Symposium; Sep 13-15, 2018; Seattle, WA. Philadelphia (PA): AACR; Clin Cancer Res 2019;25(22 Suppl):Abstract nr TMIM-065.

e17571 Background: Squamous cell carcinoma of the head and neck (SCCHN) is currently the sixth most common cancer in the world and is linked to tobacco, alcohol and human papillomavirus (HPV). Cisplatin (Cs)-based concurrent chemoradiation is currently the standard treatment for locally advanced disease with a desire for organ preservation. In 2006, cetuximab (Cx) with radiation was approved as another option for this indication. We performed a single institute retrospective analysis to explore the difference in efficacy between Cs and Cx regimens with respect to P16 and smoking status. Methods: We retrospectively reviewed pts in Loma Linda University Medical Center (LLUMC) with locally advanced SCCHN who received concurrent chemoradiation with either Cx or Cs as first-line treatment from 2006 to present. We excluded patients with nasopharyngeal cancers, and patients having surgery as first line of treatment. Overall survival (OS) and disease free survival (DFS) with respect to p16 status and smoking status are the two primary endpoints. Results: Based on our study criteria, 115 out of 1545 screened pts qualified. The average age at diagnosis was 60 years. 75.7% were males and 24.3% were females. Median follow-up is 26 months. There are overall 55 DFS events and 23 OS events. Comparing between smokers (S, n = 44) and non-smokers (NS, n = 42), there is no significant difference in OS (NS: HR = 0.73 [0.28-1.92], p = 0.52) while the DFS was significantly better in the NS (NS: HR = 0.53 [0.29-0.98], p = 0.043). The overall median OS of the P16-pos pts (n = 49) is not reached while that of P16-neg pts (n = 12) is 36 mo (P16-neg: HR = 6.431 [1.71-24.13], p = 0.0058). The overall median DFS of the P16-pos pts is not reached while that of P16-neg pts is 6.5 mo (P16-neg: HR = 5.39 [2.32-12.54], p < 0.001). Overall, the median OS of Cx is 132 mo while that of Cs is not reached (Cs: HR = 0.38 [0.16-0.90], p = 0.027). The median DFS is 30 mo for Cs and 25 mo for Cx. Within P16-pos population, there are 4 OS events in Cx (n = 18) while 0 in Cs (n = 31)(p = 0.0062, favors Cs); there are 5 DFS events in Cx while 9 in Cs (Cs: HR = 1.17 [0.39-3.51], p = 0.78). The median OS and DFS for both groups are not reached. Conclusions: Our analysis of retrospective single institute data shows that positive P16 significantly associates with better OS. These findings are consistent with previous reports. Compared to Cx, the Cs regimen significantly associates with a better OS but not DFS in both overall and P16-pos populations.

e13130Background: Age at diagnosis is an important prognostic factor for breast cancer in general however its specific significance in triple negative breast cancer (TNBC) is unclear. Few existing ...