bluuebunny/crossref_metadata_2025 huggingface dataset:
Abstract 4003: Colon sessile serrated polyps associate with endometrial polyps
Authors: Brim Hassan , Tarjoman Taraneh , Ganjali Shirin , Azam Saman , Farsi Negin , Lee Edward , Shokrani Babak , Aduli Farshad , Williams Carla , Laiyemo Adeyinka , Soleimani Akbar , Sherif Zaki , Nouraie Mehdi syed , Ashktorab Hassan
Published: August 2019

Background: Colon and endometrial polyps are current health issues and their incidences are on the rise. Due to conflicting reports of higher colon polyps in patients with endometrial polyps, we aimed to evaluate if there is such an association in African Americans, a population at high risk for colorectal cancer.

Method: We retrospectively reviewed all female patients who had endometrial (EN) presentations (n=3,600) and colonoscopy (n=14,888) at Howard University Hospital from January 2004 to December 2015. We selected cases with endometrial poly and control without this diagnosis who underwent colonoscopy. Clinical [Body Mass Index (BMI), diabetes (DM), hypertension (HTN), Tamoxifen intake, age] and pathological (histology, location, type of lesions) features of Cases and Controls were collected. Association between EN and colon polyp was tested using multivariable logistic regression analysis. We excluded the patients with endometrial cancer.

Results: We recruited 664 cases and 118 controls. Cases were statistically significant older (median age of 60 vs. 57 in controls) and had higher rates of smoking and DM than controls. However, the overall colon polyp prevalence in the two groups was not statistically different (54% vs 52% in controls and cases, respectively). In subgroups of subject with colon polyp, sigmoid and rectal polyps were more prevalent in controls than cases. Sessile serrated polyps/adenomas (SSPs) were more frequent in cases (8% vs. 2%, p=0.03) while benign mucosa was more so in controls. Whether considering the overall study population or just those with colon polyps, SSPs lesions associated with an Odds Ratio of 4.6 (p=0.02) for EN polyp occurrence, after adjusting for confounder.

Conclusion: Our study shows that the overall colon polyp’s prevalence was similar in endometrial polyp patients compared to endometrial-polyp free controls. However, a significant association between SSP with endometrial polyps was noted. Females with colon lesions of the SSP type might benefit from a screening for endometrial polyp in an age-independent manner.

Note: This abstract was not presented at the meeting.

Citation Format: Hassan Brim, Taraneh Tarjoman, Shirin Ganjali, Saman Azam, Negin Farsi, Edward Lee, Babak Shokrani, Farshad Aduli, Carla Williams, Adeyinka Laiyemo, Akbar Soleimani, Zaki Sherif, Mehdi syed Nouraie, Hassan Ashktorab. Colon sessile serrated polyps associate with endometrial polyps [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 4003.
Abstract 4006: Effect of Prior Cancer on Trial Eligibility and Treatment Outcomes in Nasopharyngeal Carcinoma: Implications for Clinical Trial Accrual
Authors: Wang Ya-Qin , Lv Jia-Wei , Tang Ling-Long , Du Xiao-Jing , Chen Lei , Li Wen-Fei , Liu Xu , Guo Ying , Lin Ai-Hua , Mao Yan-Ping , Sun Ying , Chen Yu-Pei , Ma Jun
Published: August 2019

Background: In cancer trials, prior cancer is a common exclusion criterion. We evaluated the characteristics of prior cancer exclusion criteria in nasopharyngeal carcinoma (NPC) trials and determined its prognostic effect on patients with NPC.

Methods: We reviewed NPC trials for prior cancer exclusion criteria. Then we estimated the effect of prior cancer among NPC patients using the Surveillance, Epidemiology, and End Results database. Propensity score-matching was used to compensate for differences in baseline characteristics between patients with and without prior cancer.

Results: There were 109 clinical trials involving 10,437 patients; 49 trials (45%) excluded patients with prior cancer. Prior cancer exclusion was more common in recent or phase III trials. We identified 10,195 NPC patients; 6.2% had prior cancer. More than 70% of these cancers were in situ/localized/regional and diagnosed relatively close to the NPC diagnosis (median 3.3 years). Patients with certain prior cancer type (prostate, breast, gynecological, hematological), time of diagnosis (>5 years ago), or stage (in situ/localized) did not have inferior survival compared with patients with no prior cancer. We tested one form of prior cancer exclusion criteria in an NPC cohort resembling a modern trial population: it did not adversely affect overall and NPC-specific survival.

Conclusions: Many NPC trials excluded patients with prior cancer, which impacts trial accrual and generalizability. Our findings suggest that broader inclusion in trials of patients with NPC with prior cancer might not affect trial outcomes. More research is needed to understand the appropriateness of this exclusion policy across cancer types and trials.

Citation Format: Ya-Qin Wang, Jia-Wei Lv, Ling-Long Tang, Xiao-Jing Du, Lei Chen, Wen-Fei Li, Xu Liu, Ying Guo, Ai-Hua Lin, Yan-Ping Mao, Ying Sun, Yu-Pei Chen, Jun Ma. Effect of Prior Cancer on Trial Eligibility and Treatment Outcomes in Nasopharyngeal Carcinoma: Implications for Clinical Trial Accrual [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 4006.
Abstract 4008: Association of lung cavitation development with apatinib treatment outcomes in advanced gastric and NSCLC patients with multichemotherapy failure
Authors: Jiang Man , Zhang Chuantao , Liu Dong , Zhou Na , Wang Yongjie , Hou Helei , Li Tianjun , Lv Hongying , Zhu Jingjuan , Zhang Chuanyu , Zhang Xiaochun
Published: August 2019

Purpose:

Patients who receive antiangiogenic agents commonly develop cavitation in their lung lesions. This symptom has also been found in apatinib-treated patients with lung tumors. The present study evaluated the frequency and clinical outcomes of patients who developed tumor cavitation following apatinib treatment.

Experimental Design:

This was a retrospective study of 189 patients with lung tumors who were treated with 250 mg per day of apatinib between February 1, 2015, and May 19, 2017. Clinical data were retrieved from their medical records, and chest imaging findings were documented. Survival data were analyzed with Kaplan-Meier estimates and were compared with a log-rank test. OR values were analyzed for the effect factor of cavitation development.

Results:

Cavitation development showed a benefit for patients on apatinib therapy regardless of whether they had primary or metastatic lung cancer. For patients without cavitation, metastatic lung cancer had greater 6-month LRC, PFS and OS rates than did primary lung cancer. No significant association was found between histological type and survival analysis in apatinib-treated NSCLC patients. Increased CEA levels promoted PFS in patients on apatinib who developed cavitation. However, changes in CEA values did not predict LRC, OS or PFS in patients who developed no cavitation. OR analysis showed that increased CEA levels were a risk factor for cavitation development.

Interpretation:

This was a retrospective real-world study approved by the ethics committee of the Affiliated Hospital of Qingdao University. Patients with advanced or metastatic gastric cancer and NSCLC who had progressed or relapsed after undergoing at least two lines of systemic therapy in accordance with NCCN guidelines and had never received antiangiogenic therapy were included.

Conclusions:

Lung cavitation development is common with apatinib therapy and is a potential prognostic biomarker. CEA is not a prognostic biomarker but is a prospective biomarker for lung cavitation in apatinib-treated patients.

Note: This abstract was not presented at the meeting.

Citation Format: Man Jiang, Chuantao Zhang, Dong Liu, Na Zhou, Yongjie Wang, Helei Hou, Tianjun Li, Hongying Lv, Jingjuan Zhu, Chuanyu Zhang, Xiaochun Zhang. Association of lung cavitation development with apatinib treatment outcomes in advanced gastric and NSCLC patients with multichemotherapy failure [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 4008.
Abstract 4009: GLUT3 variant is associated withprognosis of patients with non-small cell lung cancer after surgical resection
Authors: Lee Shin Yup , Do Sook Kyung , Choi Sun Ha , Choi Jin Eun , Kang Hyo Gyoung , Hong Mi Jeong , Baek Dong Won , Yoo Seung Soo , Son Ji Woong , Park Jae Yong
Published: August 2019

This study was conducted to explore whether polymorphisms of glucose transporter 3 (GLUT3) gene affect the prognosis of patients with non-small cell lung cancer (NSCLC) after surgical resection. Four single nucleotide polymorphisms (SNPs) in GLUT3 were investigated in a total of 782 patients with NSCLC who underwent curative surgery. The association of the SNPs with overall survival (OS) and disease free survival (DFS) was analyzed. Among the four SNPs investigated, GLUT3 rs7309332C>T was significantly associated with OS and DFS in multivariate analyses. The SNP was associated with significantly worse OS (adjusted hazard ratio [aHR] = 1.62, 95% confidence interval [CI] = 1.04-2.53, P = 0.03, under recessive model), and worse DFS (aHR = 1.64, 95% CI = 1.18-2.29, P = 0.003, under recessive model). When stratified by tumor histology, the association between the GLUT3 rs7309332C>T and OS/DFS was not limited to either squamous cell carcinoma (SCC) or adenocarcinoma (AC), although the significant association remained only in AC for OS (P = 0.40 for SCC and P = 0.04 for OS) and only in SCC for DFS (P = 0.03 for SCC and P = 0.08 for OS). When AC patients were stratified according to EGFR mutation status, the SNP was significantly associated with DFS in patients with EGFR mutant tumors (aHR = 2.47, 95% CI = 1.15-5.30, P = 0.02, under recessive model), but not in those with EGFR wild-type tumors. This study suggests that genetic variation in GLUT3 may be useful in predicting survival of patients with early stage NSCLC.

Citation Format: Shin Yup Lee, Sook Kyung Do, Sun Ha Choi, Jin Eun Choi, Hyo Gyoung Kang, Mi Jeong Hong, Dong Won Baek, Seung Soo Yoo, Ji Woong Son, Jae Yong Park. GLUT3 variant is associated withprognosis of patients with non-small cell lung cancer after surgical resection [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 4009.
Abstract 4019: High TERT expression in NSCLC is associated with poor prognosis
Authors: Li Tianhong , Ma Weijie , Li Qianping , Wei Sixi , Wang Tao , Xiang Jingyi , Liu Cheng
Published: August 2019

Background: Activation of human telomerase reverse transcriptase (TERT) has been linked to tumorigenesis and aggressive tumor behavior. This study was undertaken to evaluate if TERT expression was correlated with clinicopathological parameters and survival in patients with non-small cell lung cancer (NSCLC).

Methods: TMA slides made from 254 NSCLC patients (cohort 1 and cohort 2), including 127 adenocarcinoma (LUAD), 78 squamous carcinomas (LUSC) and 46 other types, were included. TMA Formalin-fixed and paraffin-embedded (FFPE) tissue sections were stained using the immunohistochemistry stain (IHC). The percentage of positive nuclei was determined for each case and the cut-off value was set as 5%, 20% and 50% positive tumor cells. TERT expression levels were analyzed for associations with patient survival rates and clinicopathological parameters (such as age, gender, tumor grade, tumor stage, metastasis). Survival analysis was performed using the Kaplan-Meier method.

Results: TERT was positive in 199/254 (78%) of the samples. High TERT expression was seen in tumor specimens compared with normal adjunct tissues (p < 0.001). Kaplan-Meier curves indicated that high TERT expression was significantly associated with poor overall survival in NSCLC patients (p < 0.001 and p < 0.0001, 20% and 50% cut-off, respectively). Multivariate analysis of survival demonstrated that TERT expression at 50% cutoff was an independent prognostic factor for NSCLC patients (HR = 0.1461, 95% CI: 0.0862-0.2475, p < 0.0001). High TERT expression was also associated with short survival of LUSC and LUAD patients at 50% cutoff (p < 0.0001). No significant association with OS was observed at 5% cutoff. There was also significant correlation between TERT expression with metastasis but no correlation with clinical stage, histological type and tumor differentiation.

Conclusion: Our study showed that high TERT expression is present in 78% of NSCLC tumors and is associated with poor prognosis. TERT may serve as a prognostic biomarker and novel drug target for NSCLC.

Citation Format: Tianhong Li, Weijie Ma, Qianping Li, Sixi Wei, Tao Wang, Jingyi Xiang, Cheng Liu. High TERT expression in NSCLC is associated with poor prognosis [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 4019.
Abstract 4028: Tenascin C in gastric cancer stroma is a predictive marker for postoperative prognosis
Authors: Kikuchi Hirotoshi , Murakami Tomohiro , Kawata Sanshiro , Hirotsu Amane , Matsumoto Tomohiro , Ozaki Yusuke , Hiramatsu Yoshihiro , Kamiya Kinji , Morita Yoshifumi , Sakaguchi Takanori , Konno Hiroyuki , Takeuchi Hiroya
Published: August 2019

Background: Tenascin C, a glycoprotein in the extracellular matrix is reportedly associated with poor clinical outcome in several malignancies. We have previously reported that tenascin C in colorectal cancer stroma is a predictive marker for liver metastasis (Br J Cancer 117:1360-70, 2017). In this study, we evaluated the clinicopathological and prognostic significance of tenascin C expression in gastric cancer stroma.

Materials and methods: Expression levels of tenascin C protein were immunohistochemically analyzed in 176 primary gastric cancer samples. Tenascin C staining intensity and relativity were classified into four scores and two groups, respectively. The correlation of the tenascin C staining intensity and relativity with clinicopathological factors, postoperative overall survival (OS) and disease-free survival (DFS) were analyzed.

Results: There were positive correlations between the tenascin C staining intensity or relativity and tumor depth, lymphatic invasion, venous invasion, lymph node metastasis, pathological stage, high preoperative carcinoembryonic antigen (CEA)>5ng/ml, and high preoperative carbohydrate antigen (CA19-9) >37U/ml. Multivariate analysis revealed that high tenascin C staining intensity significantly correlated with tumor invasion into the muscularis propria (p=0.015) and venous invasion (p=0.042), and that high tenascin C staining relativity correlated with venous invasion (p=0.013). Gastric cancer patients with high tenascin C staining intensity had significantly shorter postoperative OS (p<0.001), whereas tenascin C staining relativity did not correlate with postoperative OS (p=0.05). High tenascin C staining intensity and relativity significantly correlated with shorter postoperative DFS (p<0.001 and 0.005, respectively). Cox proportional hazard model identified high tenascin C staining intensity as an independent prognostic factor for poor postoperative OS (hazard ratio (HR): 13.16, 95% confidence interval (CI): 1.362-125.000, p=0.026) and DFS (HR: 6.02, 95%CI: 1.538-23.810, p=0.010).

Conclusions: Tenascin C protein expression in primary tumor stroma was identified as a predictive marker for postoperative prognosis in patients with gastric cancer.

Citation Format: Hirotoshi Kikuchi, Tomohiro Murakami, Sanshiro Kawata, Amane Hirotsu, Tomohiro Matsumoto, Yusuke Ozaki, Yoshihiro Hiramatsu, Kinji Kamiya, Yoshifumi Morita, Takanori Sakaguchi, Hiroyuki Konno, Hiroya Takeuchi. Tenascin C in gastric cancer stroma is a predictive marker for postoperative prognosis [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 4028.
Abstract 4035: Protein tyrosine kinase 7 (PTK7) biomarker analysis in patients (pts) treated with PF-06647020, a PTK7 antibody-drug conjugate (ADC), in a phase I dose expansion study
Authors: Jackson-Fisher Amy , Mehra Navi , Gianani Roberto , Whalen Pamela , Vizcarra Pamela , Deng Shibing , McComish Stephen , Xin Xiaohua , Powell Eric L.
Published: August 2019

Background: PF-06647020, an ADC comprising a humanized monoclonal antibody against PTK7, a cleavable valine-citrulline linker, and an auristatin payload, is being investigated in an ongoing Phase I clinical trial in pts with advanced solid tumors. We hypothesized that response to a PTK7-directed ADC would correlate with PTK7 expression in the pts tumor. We report preliminary baseline PTK7 expression in pts tumors and clinical response using a novel CLIA validated laboratory developed test (LDT).

Methods: We developed and validated an LDT comprising an anti-PTK7 immunohistochemistry assay with digital tissue analysis using Flagship Biosciences Inc’s cTA® platform to detect membrane-associated PTK7 in formalin-fixed paraffin embedded (FFPE) epithelial malignancies of ovary, breast and lung. Tumor samples were immunolabeled with the LDT and image analysis was applied to derive a digital H-score. Specifically the digital image annotation included desired regions of analysis (ROA) and excluded regions not of interest. The cTA® platform identified each cell in a given ROA and quantified an overall H-score for malignant epithelial cell membranes immunolabeled for PTK7. Non-small cell lung cancer (NSCLC) pts with H-scores ≥ 56.8 were eligible for enrollment whereas ovarian cancer (OVCA) pts were enrolled unselected. Triple negative breast cancer (TNBC) pts were initially enrolled unselected, but after review a requirement for H-scores ≥ 92.8 was implemented for enrollment.

Results: PTK7 IHC was performed on baseline FFPE samples from 69 pts and digital H-scores were generated (Table 1). An additional 7 pt samples were not amenable to analysis with the cTA® platform.

Conclusions: A novel CLIA validated LDT was developed and implemented to assess PTK7 baseline levels in FFPE tumors from pts treated with PF-06647020. Clinical responses tended to correlate with H-scores that were higher than the mean for each tumor type.

Table 1.PTK7 digital H-scores in Phase I dose expansionNumber of SamplesMinimum H-scoreMaximum H-scoreMean H-scoreMean H-score of Non-respondersMean H-score of RespondersOVCA31120089.280.2111.2NSCLC1993287167.8159.2200.3TNBC1953258159.2147.0224.3

Citation Format: Amy Jackson-Fisher, Navi Mehra, Roberto Gianani, Pamela Whalen, Pamela Vizcarra, Shibing Deng, Stephen McComish, Xiaohua Xin, Eric L. Powell. Protein tyrosine kinase 7 (PTK7) biomarker analysis in patients (pts) treated with PF-06647020, a PTK7 antibody-drug conjugate (ADC), in a phase I dose expansion study [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 4035.
Abstract 4042: Longitudinal monitoring of neoepitope-specific T cell repertoires in patient blood following cancer immunotherapy
Authors: Peng Songming , Yuen Benjamin , Tan Joanne , Yin Fangfang , Bao Robert , Pan Zheng , Dalmas Olivier , An Duo , Quach Boi , Yi Michael , Bethune Michael , Mandl Stefanie , Walters Matt , Jaen Juan , Franzusoff Alex
Published: August 2019

T cells targeting neoepitopes derived from mutations exclusive to the tumor are one of the main drivers of cancer immunotherapy efficacy. Tracking these neoepitope-specific T cells during cancer immunotherapy has been hampered by the impracticality of repeated sampling from the tumor, and by the low frequency of neoepitope-specific T cells in peripheral blood. An ultra-sensitive and high-throughput technology (imPACT) has been developed for the identification and isolation of neoepitope-specific T cells from peripheral blood. Subjects with colorectal cancer, endometrial adenocarcinoma, nasopharyngeal carcinoma and other solid tumors were treated with AB122 (anti-PD-1 antibody) as part of an ongoing dose-escalation clinical trial. Pre-treatment blood samples were analyzed to identify the basal repertoire of neoepitope-specific T cells. Evolution of this repertoire during AB122 treatment was monitored to enable immune phenotyping and correlation with clinical outcomes. In addition, transcriptional profile changes were monitored at the single-cell level for each neoepitope-specific T cell. These data will enable us to analyze T cells targeting neoepitopes and identify driver mutations that correlate with and may be responsible for therapeutic benefit. More broadly, this platform technology promises to significantly advance our understanding of T cell-mediated mechanisms of cancer immunotherapy.

Citation Format: Songming Peng, Benjamin Yuen, Joanne Tan, Fangfang Yin, Robert Bao, Zheng Pan, Olivier Dalmas, Duo An, Boi Quach, Michael Yi, Michael Bethune, Stefanie Mandl, Matt Walters, Juan Jaen, Alex Franzusoff. Longitudinal monitoring of neoepitope-specific T cell repertoires in patient blood following cancer immunotherapy [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 4042.
Abstract 4043: Application of high complexity flow cytometry for Monitoring activated T-cells, T and B regulatory subsets in combination immunotherapy trials, melanoma case study
Authors: Tsau Jennifer , Abadier Michael , Atzmiller Brittney , Vaupel Christine , Dakappagari Naveen , Sarikonda Ghanashyam , Tarhini Ahmad
Published: August 2019

Background

Ever increasing knowledge of the interplay between immune composition and a novel drugs’ treatment efficacy or the lack thereof leads to continual advancements in immunotherapies. Given the exponential increase in combination therapies, tools to determine the most promising regimens are critical.

Immunotherapeutic agents either reverse T-cell inhibitory pathways (e.g., PD-1) or activate the immune system to induce cytotoxic T-cell activation and proliferation through agonistic stimulation (e.g., Cytokines). These treatment strategies have proven to be efficacious by enhancing the balance between immune-effector and suppressive/regulatory cells that can directly affect the outcome of immunotherapy.

To concurrently monitor multiple immune cell subsets responsible for either immuno-suppression or activation, we developed a high-complexity (13-color) flow cytometry panel to reliably enumerate activated T-cells, Tregs (A & B subsets) and Bregs for clinical trial application.

Methods

The panel includes CD3, CD4, CD8, CD19, FoxP3, CD25, CD127, CD45RA, HLA-DR, CD38, CD24, and CD27 to monitor total Tregs, Treg-subsets, Bregs and T-cell activation. After robust analytical method validation following industry guidelines for exploratory biomarker assessments, we confirmed the clinical utility of this panel in stage III or IV melanoma patients (N= 14) enrolled in a randomized phase II clinical trial receiving high-dose of IL-2 with or without Afliberecept (VEGF inhibitor).

Results

We observed up-to 20-fold increase in activated CD8+ T-cells following treatment. Interestingly, while no discernible change was observed in the naive Treg-A cells (CD45RAhiFoxp3lo), a remarkable 5-fold expansion of effector Treg-B cells (CD45RAloFoxp3hi) was observed following treatment. Additionally, frequencies of Bregs (CD24hiCD38hi) remained unchanged after treatment.

Conclusions

In summary, we demonstrated the utility of a powerful high complexity clinical flow cytometry tool for precisely monitoring immune cell subsets involved in tumor escape such as Tregs and Bregs or those involved in tumor killing like CD8+ cytotoxic T-cells. More notably, this tool may be utilized to deprioritize agents that cause immunosuppression and focus on combination agents that lead to additive immune activation.

Citation Format: Jennifer Tsau, Michael Abadier, Brittney Atzmiller, Christine Vaupel, Naveen Dakappagari, Ghanashyam Sarikonda, Ahmad Tarhini. Application of high complexity flow cytometry for Monitoring activated T-cells, T and B regulatory subsets in combination immunotherapy trials, melanoma case study [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 4043.
Abstract 4046: Immune repertoire sequencing reveals tumor microenvironment and tracks clonally expanded B cell and T cell in blood
Authors: Song Chen , Liu Pingfang , Barry Andrew , Langhorst Bradley W. , Stewart Fiona J. , Russello Salvatore , Davis Theodore B. , Dimalanta Eileen T.
Published: August 2019

The study of complex immunological diseases and tumor microenvironments has progressed through recent developments on sequencing of the immune repertoire. Using this approach, the interrogation of disease progression is facilitated through analysis of millions of V(D)J combinations from B cell antibodies (Igs) and T cell receptors (TCRs). One major challenge of immune repertoire sequencing is to accurately capture the structural and sequence complexities of antibodies and TCR genes. We have developed a method for accurate sequencing of full length immune gene repertoires of B cells and T cells. RNA extracted from tumor tissues containing tumor infiltrating lymphocytes, as well as matched peripheral blood mononuclear cells (PBMCs) were used to generate full length Ig and TCR libraries. Unique molecule index (UMI) was used to discretely barcode each mRNA molecule, enabling absolute quantitative ranking of antibody/TCR clone abundance. Full length immune repertoire sequencing facilitates detection of distinct and shared clones in tissue and blood samples, enabling identification of disease specific clones to evaluate immunotherapy effects. Highly expanded clones in tumor samples are also found in blood samples. RNA-seq libraries were also constructed from the same RNA for Ig and TCR libraries. The expression level of IGH/IGL/IGK and TRA/TRB in the immune sequencing libraries highly correlates with the RNA-seq data. In addition, both Ig and TCR libraries can be constructed in one tube to obtain a whole immune repertoire profile.Our immune repertoire sequencing approach allows accurate clonal determination for both Ig and TCR. This technique is applicable for investigation of lymphocytes infiltration of tumor microenvironments, tracing expanded B cell and T cells in blood samples, and monitoring of minimal residual disease.

Citation Format: Chen Song, Pingfang Liu, Andrew Barry, Bradley W. Langhorst, Fiona J. Stewart, Salvatore Russello, Theodore B. Davis, Eileen T. Dimalanta. Immune repertoire sequencing reveals tumor microenvironment and tracks clonally expanded B cell and T cell in blood [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 4046.
Abstract 4048: REAlease® technology: Controlled release of antibody-fluorochrome conjugates for maximal flexibility in flow sorting and fluorescence microscopy applications
Authors: Pankratz Jennifer , Schmachtenberg Sabine , Jansen Nicole , Hansen Marie , Krauthäuser Susanne , Kinkhabwala Ali , Schlegel Karolina , Meiler Svenja , Siewert Christiane , Bartholomäus Alina , Yushchenko Dmytro A. , Dose Christian
Published: August 2019

Multi-parameter immunofluorescent labeling is the method of choice for the flow-based sorting of targeted cell populations from heterogeneous samples. However, existing antibody-fluorochrome conjugates used for flow sorting limit the types of downstream analyses that can be applied to the isolated cells, as their continued presence on the cells blocks specific cellular epitopes and restricts the choice of downstream fluorescence detection channels. Herein, we present a new type of antibody-fluorochrome conjugates that facilitates a highly specific multi-parameter cell staining for flow sorting as well as subsequent complete release of the conjugate probes from the sample. These new conjugates rely on recombinantly engineered antibody fragments that individually possess low epitope binding affinities. A tailor-made covalent conjugation chemistry allows for their multimerization as well as fluorochrome labeling, generating fluorescent probes comparable to conventional antibody conjugates. Importantly, upon addition of a release reagent, the antibody-fluorophore conjugates can be rapidly released from the cell surface. Accordingly, previously blocked epitopes are free for re-labeling, which provides maximal flexibility for renewed epitope targeting in downstream applications. As many targeted cell populations require multi-parameter detection of several antigens for their isolation, the recombinant engineered antibodies were conjugated with different fluorochromes. The versatility of this technology is presented in multi-parameter panels enabling clear distinction and discrimination of different cell populations of different starting material, e.g. peripheral blood and tumor cells. The applicability of the new REAlease® antibody fluorochrome conjugates is also demonstrated by flow sorting of human naïve regulatory T cells. In addition, we show that the fast and efficient releasability of the new conjugates can serve as an optimal basis for cyclic immunofluorescence microscopy using a new MACSima™ imaging platform, which permits cell analysis with potentially unlimited number of parameters.

Citation Format: Jennifer Pankratz, Sabine Schmachtenberg, Nicole Jansen, Marie Hansen, Susanne Krauthäuser, Ali Kinkhabwala, Karolina Schlegel, Svenja Meiler, Christiane Siewert, Alina Bartholomäus, Dmytro A. Yushchenko, Christian Dose. REAlease® technology: Controlled release of antibody-fluorochrome conjugates for maximal flexibility in flow sorting and fluorescence microscopy applications [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 4048.
Abstract 4054: Mass cytometry approaches to biomarker discovery via high-dimensional antigen-specific T cell identification and profiling
Authors: Cobbett Mathew , Fehlings Michael , Newell Evan
Published: August 2019

immunoSCAPE leverages the high-dimensional immune profiling capabilities of mass cytometry combined with a unique technology for the identification of antigen-specific T-cells to support the development of immunotherapy strategies in cancer and immune-related diseases. In cancer, there is now strong evidence that immunotherapy-mediated tumor rejection can rely on tumor-specific (neoantigen-specific) CD8+ T-cells. Consequently, the discovery of neoantigens becomes valuable for personalized cancer immunotherapies. Although in silico pipelines exist, that are capable of predicting non-synonymous mutations potentially giving rise to tumor-specific neoantigens, it is not clear how accurate these methods are in identifying immunogenic and therapeutically relevant epitopes, since T-cell epitope usage can be influenced by many factors. Moreover, analysis of T-cells in cancer patients is challenging as it requires detecting rare antigen-specific T-cell populations in samples that are usually limited in volume and availability.

By applying cytometry by time of flight in conjunction with combinatorial peptide-MHC tetramer staining and high-performance dimensional analysis tools, we are able to map broadly MHC-class I epitope with a high sensitivity for rare antigen-specific T-cells and perform concurrently in-depth characterization of these cells. We will show here the application of this technology in the context of immunotherapy, through the example of a murine in vivo tumor model responsive to checkpoint blockade inhibitors, as well as through the analysis of different human cancer samples.

Together, by providing insights into the nature of neoantigen-specific T-cells, immunoSCAPE’s unique target discovery and high-dimensional immune profiling platform is a valuable tool for the development of novel diagnostic biomarkers and therapeutic strategies at different stages of drug development.

Citation Format: Mathew Cobbett, Michael Fehlings, Evan Newell. Mass cytometry approaches to biomarker discovery via high-dimensional antigen-specific T cell identification and profiling [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 4054.
Abstract 4073: Novel BETi NHWD-870 synergizes with immune checkpoint inhibitor in preclinical model of non-small cell lung cancer
Authors: Zhang Yongchang , Sun Ziyi , Yang Nong
Published: August 2019

Background: Small molecular inhibitors targeting bromodomain and extraterminal domain (BET) protein is promising in cancer therapy. NHWD-870 was a highly selective BETi and has been proved effective in lung cancer according to our previous findings. Combination of immune checkpoint inhibitors with BETi is now under investigation for lung cancer treatment.

Method: We set up a fast track preclinical model with C57 mouse. Growth rate and survival data were measured in subgroups receiving single NHWD-870, single PD-1 antibody and combined treatment respectively. Finally, pathology sections, immunohistochemistry and inflammatory cell infiltration was applied to evaluate the pathological response.

Result: Millions of mouse lung cancer cells were harvested and injected to the lateral leg. Lung cancer xenografts were visible in 3-5 days. And the total survival time was within 30 days without any treatment. Tumor volume was measured for effective evaluation. During the three groups, NHWD-870 combined with anti-PD-L1 showed robust efficacy in inhibiting tumor growth. And the survival time was also prolonged significantly. Pathological examination showed the lung cancer cells were filled with necrosis. Inflammatory cell infiltration and immunohistochemistry showed that PD-L1 was down regulated and CD4+/ CD8+ positive lymphocyte cells increased significantly.

Conclusion: We established a useful method to evaluate tumor response to immune checkpoint inhibitor. And our data proved BETi plus immune checkpoint inhibitor to be a promising treatment of NSCLC.

Citation Format: Yongchang Zhang, Ziyi Sun, Nong Yang. Novel BETi NHWD-870 synergizes with immune checkpoint inhibitor in preclinical model of non-small cell lung cancer [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 4073.
Abstract 4077: ATOR-1144 is a tumor-directed CTLA-4 x GITR bispecific antibody that acts by depleting Tregs and activating effector T cells and NK cells
Authors: Fritzell Sara , Levin Mattias , Åberg Ida , Johansson Maria , Winnerstam Magnus , Smith Karin Enell , Ellmark Peter , Furebring Christina , Norlén Per , Kvarnhammar Anne
Published: August 2019

ATOR-1144 is a human CTLA-4 x GITR bispecific IgG1 antibody generated by fusing a high affinity CTLA-4 binder, derived by FIND® optimization of the CTLA-4 binding domain of CD86, to an agonistic GITR antibody isolated from the human antibody library ALLIGATOR-GOLD®. CTLA-4 and GITR are highly expressed on Tregs in the tumor microenvironment, and GITR is also expressed on NK cells and certain tumor cells. ATOR-1144 was designed to induce a tumor-directed immune activation for treatment of solid tumors and hematological malignancies. The in vitro activity of ATOR-1144 in terms of Treg depletion and activation of T cells and NK cells was investigated using purified cells from healthy human donors. ATOR-1144 induced depletion of primary Tregs in an ADCC assay with NK cells. T-cell activation in terms of IL-2 and IFN-γ release was demonstrated in the presence of CTLA-4 and Fcγ receptor crosslinking. NK cells treated with ATOR-1144 released IFN-γ, perforin and granzyme B, and displayed enhanced cytolytic killing of tumor cells. Moreover, NK cell-mediated depletion of CTLA-4-expressing cells was significantly improved upon stimulation with ATOR-1144 compared to monospecific anti-CTLA-4, indicating that NK cells activated via GITR acquire an enhanced capacity to induce ADCC. In conclusion, ATOR-1144 is a first-in-class bispecific tumor directed antibody targeting CTLA-4 and GITR. ATOR-1144 acts through several mechanisms, including depletion of Tregs, activation of effector T cells and activation of NK cells for enhanced Treg depletion and tumor cell killing.

Citation Format: Sara Fritzell, Mattias Levin, Ida Åberg, Maria Johansson, Magnus Winnerstam, Karin Enell Smith, Peter Ellmark, Christina Furebring, Per Norlén, Anne Kvarnhammar. ATOR-1144 is a tumor-directed CTLA-4 x GITR bispecific antibody that acts by depleting Tregs and activating effector T cells and NK cells [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 4077.
Abstract 4084: Radiation enhances efficacy of TEM1-specific cancer vaccination
Authors: Pierini Stefano , Uribe-Herranz Mireia , Perales-Linares Renzo , Beghi Silvia , Costabile Francesca , Pustylnikov Sergei , Facciabene Andrea
Published: August 2019

Tumor endothelial marker 1 (TEM1 or CD248) is a protein found in both tumor vasculature and stroma. In prior experiments, we showed that immunizing with Tem1-TT plasmid-DNA (Tem1 cDNA fused to the minimal domain of the C fragment of tetanus toxoid) was effective in controlling tumor progression in three mouse tumor models in a T cell-dependent manner. Moreover, effective Tem1-TT vaccination reduced tumor vascularity without impacting normal angiogenesis. Radiation therapy (RT) is an established curative and palliative cancer treatment regimen which uses high-energy radiation to kill cancer cells. RT modulates the tumor microenvironment - by increasing MHC-I expression, inducing immunological cell death, and damaging the tumor-associated vasculature - potentially synergizing with anti-cancer immunotherapies. Here, we combined RT and Tem1-TT heterologous vaccination and showed that combination of the two therapies was more effective in controlling CT26 and TC1 tumor progression than either single therapy alone. Interestingly adding RT to Tem1-TT vaccine resulted in a stronger vaccine-specific immune response and increased epitope spreading toward gp70, a TAA expressed by CT26 tumor cell line. Further analysis of the tumor microenvironment revealed a significant increment of PD-L1 expression in both tumor cells and tumor vasculature after single or either combination therapy. This led us to test whether blocking the PD1/PD-L1 axe would further improve the efficacy of RT plus Tem1-TT therapy. Our results demonstrate that administration of anti-PD-L1 antibody further synergized with RT plus Tem1-TT and improved mice survival. In conclusion, triple combination of Tem1-TT vaccine, RT and anti-PD-L1 can offer a therapeutic advantage for TEM1-positive tumors which also express PD-L1.

Citation Format: Stefano Pierini, Mireia Uribe-Herranz, Renzo Perales-Linares, Silvia Beghi, Francesca Costabile, Sergei Pustylnikov, Andrea Facciabene. Radiation enhances efficacy of TEM1-specific cancer vaccination [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 4084.
Abstract 4095: Adding fuel to the fire: Augmenting the efficacy of adoptive T-cell therapy with pro-oxidants
Authors: Aboelella Nada , Zhou Gang , Fesenkova Kateryna , Ding Zhi-chun
Published: August 2019

Cancer cells face increased oxidative stress driven by oncogene activation, tumor suppressor gene mutation and increased metabolism. Even though cancer cells can resist this redox imbalance by activating various antioxidant defense mechanisms, they are more susceptible to further redox disruption by pro-oxidants. Our previous study indicated that adoptive T-cell therapy (ACT) leads to heightened oxidative stress in tumor cells reflected by depletion of cellular antioxidant glutathione (GSH), accumulation of reactive oxygen species (ROS), and presence of oxidative DNA damages. Based on this observation, we propose that intensifying oxidative stress in tumor with pro-oxidants can sensitize cancer cells to ACT and lead to improved therapeutic outcomes. In this study, we used clinically relevant ACT models to screen a panel of pro-oxidant agents for their ability to potentiate the antitumor effects of ACT. We identified several clinically applicable pro-oxidants, including some commonly used non-steroidal anti-inflammatory drugs (NSAIDs), that can benefit the infusion of tumor-specific CD4+ T cells or CD19-targeting CAR T cells. Our data provide a rationale for combining selected pro-oxidants with ACT to achieve synergistic antitumor effect.

Citation Format: Nada Aboelella, Gang Zhou, Kateryna Fesenkova, Zhi-chun Ding. Adding fuel to the fire: Augmenting the efficacy of adoptive T-cell therapy with pro-oxidants [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 4095.
Abstract 4096: Clinical study of personalized neoantigen-based cancer vaccine in the treatment of advanced SCLC patients with brain metastasis: A case report
Authors: Gao Song , Li Yongchao , Ding Zhenyu , Xu Heng , Wang Jiaqian , Guo Xianling , Fang Juemin , Wei Guochao , Qin Huanlong , Wei Yuquan , Xu Qing , Yang Li
Published: August 2019

Background: Tumor neoantigens arising from somatic mutations are less likely to be subjected to central immune tolerance and therefore the attractive targets for designing cancer vaccine. The personalized neoantigen-based cancer vaccine, a passive immunotherapy based on the theory of adaptive immune responses, has been developed rapidly in recent years.

Case Presentation: A 75-year-old male small cell of lung carcinoma (SCLC) patient with brain metastasis, who have experienced multiple chemotherapy resistance, and disease relapse after PD-1 therapy, was enrolled in this study. Whole-exome sequencing (WES) and RNA-seq were used to evaluate patient’s mutation burden and to predict the potential neoantigens. Twelve neoantigen peptides were selected and then were synthesized to generate autologous dendritic cell (DC)-based neoantigen vaccine. At last, 1.5 × 108 DC-based neoantigen vaccine was subcutaneously injected into the patient’s body to evaluate the treatment efficacy. By following-up to 10 months after vaccination, we observed an objective response with all brain metastases regressed. The immunogenicity of each of the 12 neo-epitope administered in this study was analyzed by IFNγ ELISpot in CD4+ and CD8+ T cells in pre- and post-vaccination blood samples. Responses were detected against 66.67% (8/12) of the predicted neo-epitope.

Conclusions: The autologous DC-based neoantigen cancer vaccine can effectively trigger a positive immune response to patients’ tumor and prolong patients’ survival. The vaccination can also augment the anti-tumor response of T cells by targeting other nonsynonymous mutation-derived neoepitopes. Therefore, these data demonstrated that the autologous DC-based neoantigen cancer vaccine is safe and effective in eliciting a broad antitumor immunity. Moreover, our work has introduced a framework for testing neoantigen-based vaccine combinatorial strategies in patients with advanced recurrent cancers.

Citation Format: Song Gao, Yongchao Li, Zhenyu Ding, Heng Xu, Jiaqian Wang, Xianling Guo, Juemin Fang, Guochao Wei, Huanlong Qin, Yuquan Wei, Qing Xu, Li Yang. Clinical study of personalized neoantigen-based cancer vaccine in the treatment of advanced SCLC patients with brain metastasis: A case report [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 4096.
Abstract 4099: Intrinsic Tumor Cell Consequences of Acquired Resistance to PD-L1 Blockade
Authors: Shi Yuhao , Mastri Michalis , Dolan Melissa , Oberst Michael , Ebos John
Published: August 2019

The use of immune checkpoint inhibitors (ICIs) that block programmed cell death protein ligand 1 (PD-L1) binding to PD-1 have led to durable responses in multiple cancer types; however, a subset of patients eventually stop responding. Currently, few preclinical models faithfully recapitulate clinically-relevant acquired ICI resistance that could help identify second-line treatments. Here we describe the in vivo derivation of cells from orthotopically implanted mouse mammary EMT-6 primary tumors in mice that were initially responsive to PD-L1 inhibition but eventually developed resistance. Transcriptomic comparisons of in vivo-derived parental (EMT6P) and anti-PD-L1 resistant (EMT6PDR) cells revealed alterations in multiple signaling pathways controlling secretory profiles that govern metastasis and angiogenesis. Using a small molecule screen of ~1500 FDA approved cancer drugs, we found that anti-proliferative activity in EMT6PDR cells was decreased for ~120 compounds, including numerous receptor tyrosine kinase inhibitors (RTKIs) currently being tested in ICI-refractory patient populations. Molecular analysis indicated resistance-mediated secretory changes may be linked to intrinsic functions of PD-L1 on the resistant tumor cell which, in turn, may explain differential RTKI sensitivity. Subsequent studies show that EMT6PDR secretory changes can be interferon (IFN) mediated. Together, these results reveal that acquired resistance to PD-L1 blockade can result in PD-L1-regulated intrinsic tumor cell changes involving secretory proteins and controlled, at least in part, by IFN signaling. Current studies seek to assess the impact of these cellular changes on RTKI responsiveness to identify secondary treatments after ICI failure.

Citation Format: Yuhao Shi, Michalis Mastri, Melissa Dolan, Michael Oberst, John Ebos. Intrinsic Tumor Cell Consequences of Acquired Resistance to PD-L1 Blockade [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 4099.