Kathy Robinson

Immune Characterization of the Programmed Death Receptor Pathway in High Risk Prostate Cancer

Micro‐Abstract The objective of this study was to determine the expression of programmed cell death‐1 (PD‐1) and programmed cell death ligand‐L1 (PD‐L1) in high‐grade prostate cancer tissues, and correlate the expression with disease and patient characteristics. Of the 25 samples, 2 (8%) scored high for PD‐1 expression, 2 (8%) scored high for PD‐L1 expression, and 18 (72%) scored high for CD3 expression. Background: Programmed cell death‐1 (PD‐1), a T‐cell inhibitory receptor, and its ligand, PD‐L1, have been reported to be expressed in many tumor types, and this expression has led to the development of many drugs targeting the PD‐1 pathway. The objective of this study was to determine the expression of PD‐1 and PD‐L1 in high‐grade prostate cancer tissues, and correlate the expression with disease and patient characteristics. Materials and Methods: Immunohistochemistry for PD‐1 (CD279), PD‐L1 (B7‐H1), and CD3 was performed and scored from 0 to 5 on prostatectomy/biopsy tissue samples taken from 25 men with high‐grade prostate cancer. Charts were then retrospectively reviewed for numerous patient and disease characteristics. Statistical analyses were done to investigate the association of these patient and disease characteristics with PD‐1, PD‐L1, and CD3 expression. Results: A score of 3 to 5 on the semiquantitative 0 to 5 score was deemed “high” expression whereas a score of 0 to 2 was deemed “low” expression. Of the 25 samples, 2 (8%) scored high for PD‐1 expression, 2 (8%) scored high for PD‐L1 expression, and 18 (72%) scored high for CD3 expression. There was no statistically significant difference between high and low expression groups of PD‐1, PD‐L1, or CD3 for any of the variables we collected. Conclusion: An overall low expression of PD‐1 and PD‐L1, and a concurrent high expression of CD3+ T cells was found in high‐risk prostate cancer tissue. No significant association was found between expression of PD‐1, PD‐L1, or CD3, and patient or disease characteristics. Because of this, one might be able to question the role of PD‐L1 in local immune suppression in prostate cancer.

Characterization of immortalized human mammary epithelial cell line HMEC 2.6.

Primary human mammary epithelial cells have a limited life span which makes it difficult to study them in vitro for most purposes. To overcome this problem, we have developed a cell line that was immortalized using defined genetic elements, and we have characterized this immortalized non-tumorigenic human mammary epithelial cell line to establish it as a potential model system. human mammary epithelial cells were obtained from a healthy individual undergoing reduction mammoplasty at SIU School of Medicine. The cells were transduced with CDK4R24C followed by transduction with human telomerase reverse transcriptase. Post all manipulation, the cells displayed a normal cell cycle phase distribution and were near diploid in nature, which was confirmed by flow cytometry and karyotyping. In vitro studies showed that the cells were anchorage dependent and were non-invasive in nature. The cell line expressed basal epithelial markers such as cytokeratin 7, CD10, and p63 and was negative for the expression of estrogen receptor and progesterone receptor. Upon G-band karyotyping, the cell line displayed the presence of a few cytogenic abnormalities, including trisomy 20 and trisomy 7, which are also commonly present in other immortalized mammary cell lines. Furthermore, the benign nature of these cells was confirmed by multiple in vitro and in vivo experiments. Therefore, we think that this cell line could serve as a good model to understand the molecular mechanisms involved in the development and progression of breast cancer and to also assess the effect of novel therapeutics on human mammary epithelial cells.

Single nucleotide variant in Nucleoporin 107 may be predictive of sensitivity to chemotherapy in patients with ovarian cancer

Background Alterations in nuclear pore complex (NPC) genes have been previously associated with response to chemotherapy. Using agnostic exome sequencing, we envisioned that new alleles in NPC genes, predictive of sensitivity to platinum treatment, could be discovered. Methods Twenty-two platinum-sensitive and six platinum-resistant ovarian cancer patients were tested. Platinum sensitivity was defined as disease-free survival greater than 6 months. Next-generation sequencing of exomes was used to compare platinum-sensitive and platinum-resistant patients. Single nucleotide variants (SNVs) associated with platinum sensitivity in NPC genes (n=30 genes) were identified. Results SNVs in three NPC genes were associated with response to platinum on univariate analysis. SNV rs79419059 (10T>C) in Nucleoporin 107 (Nup107) was associated with platinum resistance (P=0.0061), whereas rs2302811 (3662-4A>G) in Nucleoporin 188 (Nup188) and rs77246077 (3420-67T>A) in Nucleoporin 214 (Nup214) were associated with platinum sensitivity (P=0.0483 and 0.0091, respectively). Controlling for other confounders, multivariate age-adjusted Cox proportional hazard analysis showed rs79419059 to be significantly associated with platinum resistance (odds ratio: 4.519, 95% confidence interval: 1.317–15.501, P=0.0457). Conclusion We identified a variant in the 3′-UTR region Nup107 unique to sensitivity to platinum in ovarian cancer. With validation of this variant, it is possible that a new marker predictive of patient response may be identified.

Evaluating Adaptation of a Cancer Clinical Trial Decision Aid for Rural Cancer Patients: A Mixed-Methods Approach

Rural-residing cancer patients often do not participate in clinical trials. Many patients misunderstand cancer clinical trials and their rights as participant. The purpose of this study is to modify a previously developed cancer clinical trials decision aid (DA), incorporating the unique needs of rural populations, and test its impact on knowledge and decision outcomes. The study was conducted in two phases. Phase I recruited 15 rural-residing cancer survivors in a qualitative usability study. Participants navigated the original DA and provided feedback regarding usability and implementation in rural settings. Phase II recruited 31 newly diagnosed rural-residing cancer patients. Patients completed a survey before and after using the revised DA, R-CHOICES. Primary outcomes included decisional conflict, decision self-efficacy, knowledge, communication self-efficacy, and attitudes towards and willingness to consider joining a trial. In phase I, the DA was viewed positively by rural-residing cancer survivors. Participants provided important feedback about factors rural-residing patients consider when thinking about trial participation. In phase II, after using R-CHOICES, participants had higher certainty about their choice (mean post-test = 3.10 vs. pre-test = 2.67; P = 0.025) and higher trial knowledge (mean percentage correct at post-test = 73.58 vs. pre-test = 57.77; P < 0.001). There was no significant change in decision self-efficacy, communication self-efficacy, and attitudes towards or willingness to join trials. The R-CHOICES improved rural-residing patients’ knowledge of cancer clinical trials and reduced conflict about making a trial decision. More research is needed on ways to further support decisions about trial participation among this population.

First Report of ARID1A Somatic Mutation in a Female Patient With Renal Cell Carcinoma

Among the many genetic alterations in renal cell carcinoma (RCC), changes in VHL gene on chromosome 3 (3p25 to 26) have been shown to play a major role in the pathogenesis of RCC. Another gene commonly altered in RCC is the PBRM1 gene, which has been found to be the second major gene altered in clear cell RCC. BAP1, which is active in the ubiquitin mediated proteolysis pathway, also has been found to be altered in RCC. In addition, the inactivation of histonemodifying genes, including SETD2 and the JARID1C, has been associated with a higher likelihood of developing RCC. Among the recurrently mutated genes related to chromatin remodeling, ARID1A has been identified commonly in several studies of cancers. However, somatic mutations in ARID1A in RCC have not been reported before in the literature. We report the first somatic splice site variant affecting ARID1A that has in clear cell RCC. A targeted analysis of the somatic DNA of an individual with RCC showed a c.1920 þ 1G > A variant in the ARID1A. The findings of splice site variants of ARID1A mutation could be of therapeutic benefit if these variants were proven to be contributing to RCC growth and aggressiveness.

Abstract 1460: Bone marrow-derived CD11b+/Podoplanin+ cells are lymphatic progenitors directly responsible for breast cancer lymphatic formation

Introduction: Lymphatic metastasis, a key factor for poor outcome of breast cancer (BC), strongly depends on lymphatic vessel (LV) formation. Recent studies suggest that lymphangiogenesis is strongly promoted by bone marrow (BM)-derived CD11b + monocytes that differentiate into Lymphatic Endothelial Cell Progenitors dubbed here M-LECP. While BC are known to recruit BM monocytes, a specific BM subset responsible for tumor lymphatic formation has not been identified. We recently discovered that podoplanin (Pdpn) is the highest upregulated lymphatic marker that signifies transdifferentiation of either human or mouse immature myeloid cells into M-LECP. We hypothesized that BM-derived M-LECP are represented by a subset expressing podoplanin in CD11b + myeloid cells that are capable of inducing tumor lymphatics. Methods: Tumor lymphatic vessels expressing myeloid markers were detected immunohistochemically by triple-staining in clinical specimens and experimental breast tumors. Expression of Vegfr-3, Lyve-1, Sca-1, and other markers in BM-derived CD11b + /Pdpn + and Pdpn-negative subsets was determined by FACS. Expression of lymphatic-specific markers in CD11b + /Pdpn + and Pdpn − myeloid cells isolated from MDA-MB-231 tumors was determined by RT-qPCR. CD11b + /Pdpn + and Pdpn-negative subsets from BM of GFP-expressing mice were adoptively transferred to mice with orthotopic breast tumors followed by quantifying mobilized GFP + /CD11b + /Pdpn + cells and the density of tumor LV. Results: M-LECP were absent in normal human breast tissues but highly present in tumors. Analysis of clinical BC specimens showed significant correlations between tumor-mobilized M-LECP, density of LV and metastasis to regional nodes. All examined BC models exhibited very high densities (60-90%) of double-positive macrophages expressing lymphatic markers, and lymphatic vessels expressing myeloid proteins. At least a half of CD11b + cells isolated from tumors were positive for podoplanin and nearly 90% of CD11b + /Pdpn + subset expressed markers of progenitor cells. Only CD11b + /Pdpn + subset (but not other myeloid cells) expressed a broad panel of proteins specific to the lymphatic endothelial lineage. Adoptive transfer of BM-derived CD11b + /Pdpn + or Pdpn-negative fractions into tumor-bearing mice showed that only Pdpn+ BM myeloid cells integrated into preexisting LV and caused a 10-fold increase in peritumoral and intratumoral LV density. Conclusion: We show for the first time in clinical BC samples that tumor-recruited M-LECP significantly correlate with LN metastasis. We also identified a phenotypically distinct BM fraction of CD11b + /Pdpn + cells that directly promote generation of new tumor lymphatic vessels. Our data suggest that tumors induce myeloid BM cells to undergo differentiation into M-LECP that subsequently promote cancer lymphatics thereby promoting tumor spread. Citation Format: Caitlin Griggs, Kelly Hall, Lisa Volk-Draper, Kathy Robinson, Sophia Ran. Bone marrow-derived CD11b + /Podoplanin + cells are lymphatic progenitors directly responsible for breast cancer lymphatic formation. [abstract]. In: Proceedings of the 107th Annual Meeting of the American Association for Cancer Research; 2016 Apr 16-20; New Orleans, LA. Philadelphia (PA): AACR; Cancer Res 2016;76(14 Suppl):Abstract nr 1460.

A Pilot Study of Induction Triplet Chemotherapy Followed by Minimally Invasive Surgery for Stage 3-4 Resectable Oropharyngeal Cancer

The current study reports the results of an open label pilot study evaluating a novel treatment protocol for patients with locally advanced head and neck cancer. Patients received induction chemotherapy and those who had a clinical complete response to induction chemotherapy were offered trans-oral nidusectomy for the primary lesion and neck node dissection. If a pathologic complete response was achieved, patients were then observed. Patients not achieving a clinical or pathologic complete response to induction therapy were subsequently treated with chemoradiation.