bluuebunny/crossref_metadata_2025 huggingface dataset:
Abstract 4900: Dynamic metabolic response of prostate cancer patients treated with ADT and low carb diet
Authors: Freedland Stephen , Lin Pao-Hwa , Chen Emily Y. , Tolstikov Vladimir , Chi Jen-Tsan , Sarangarajan Rangaprasad , Narain Niven R. , Kiebish Michael A.
Published: August 2019

Background: Prostate cancer (PrCa) is one of the most common cancers among men and managed through surgery, hormonal therapy, chemotherapy, radiation, and cryotherapy. Depending on the stage of PrCa, androgen deprivation therapy (ADT) is commonly utilized as an intervention. ADT has proven effective in intervening in PrCa progression, although there are several side effects. Recently, the use of low carb diets has been shown to alter patients’ metabolic phenotype and as such may reduce the side effects of ADT.

Methods: In this study 35 men, who were beginning ADT, were randomized to low carb diet intervention with recommended exercise or control (no lifestyle change) for 6 months. Primary results have been reported showing significant weight loss at both 3-month (17lb, p<0.01) and 6-month (23 lbs, p<0.01) and improved insulin control at 3-month (HOMA=-19 as compared to baseline, p=0.02) but not at 6-month (HOMA=-4 as compared to baseline, p=0.13). For this exploratory analysis, sera collected at baseline, 3, and 6 months was used for metabolomics analysis utilizing GC/MS TOF, QqQ LC-HILIC-MS/MS, and TripleTOF 6600 LC-RP-MS and lipidomics analysis using TripleTOF 5600+ MS/MSALL workflows to quantify the chemical diversity of the metabolome and lipidome.

Results: Over 450 metabolites and 1000 lipid species were quantified. Metabolomics analysis of sera from the control patients (ADT alone) demonstrated alterations in steroid biosynthesis, androgen/estrogen metabolism, fatty acid metabolism, and lysine degradation. Lipidomics analysis of the control patients demonstrated changes in selective long chain polyunsaturated phosphatidylcholine species. Analysis of the ADT plus low carb patients revealed a change in pyruvate, glucose-alanine cycle, selenoamino acid, phenylalanine/tyrosine, and taurine metabolism. Further, lipidomic analysis revealed changes in several triglycerides, plasmalenylethanolamine, and phosphatidic acid species.

Conclusion: In summary, integration of metabolomic and lipidomic analysis in hormonal and metabolic interventions of PrCa patients revealed dynamic changes providing novel insight for tailoring therapeutic intervention and monitoring.

Citation Format: Stephen Freedland, Pao-Hwa Lin, Emily Y. Chen, Vladimir Tolstikov, Jen-Tsan Chi, Rangaprasad Sarangarajan, Niven R. Narain, Michael A. Kiebish. Dynamic metabolic response of prostate cancer patients treated with ADT and low carb diet [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 4900.
Abstract 4925: Development of a novel PTK7 immunohistochemistry (IHC) assay and analytical validation in epithelial malignancies of lung, breast and ovary
Authors: Whalen Pamela M. , Jackson-Fisher Amy , Vizcarra Pamela , Painter Cory , Pirie-Shepherd Steven , Fernandez Diane R. , Lee Joseph , Powell Eric L.
Published: August 2019

An immunohistochemistry (IHC) assay was developed to detect protein tyrosine kinase 7 (PTK7) membrane staining in formalin-fixed paraffin embedded (FFPE) human tumors with the intention of selecting non-small cell lung carcinoma (NSCLC) patients, breast cancer (BrCa) patients, or ovarian cancer (OVCA) patients for enrollment in an early stage clinical trial for a PTK7-ADC (antibody drug conjugate) experimental therapeutic. The IHC assay was developed using an anti-PTK7 specific antibody and a panel of five cancer cell lines with a range of constitutive PTK7 expression as characterized by gene transcript and protein based assays. PTK7 messenger RNA (mRNA) and protein levels were highly correlative in the control cell lines in vitro and the corresponding cell line xenografts. A pre-analytic variables study (PAV) using these characterized control cell line xenografts demonstrated that the PTK7 IHC assay was robust to the pre-analytic variables of under-fixation and ischemia. This IHC assay was used to profile PTK7 levels in human lung cancers, breast cancers, and ovarian cancers, and a dynamic range of PTK7 H-scores was detected (see Table 1). The resulting PTK7 H-scores correlated with orthogonal methods for measuring PTK7 protein and gene transcript levels in the human tumors.

Table 1.PTK7 H-scoresNumber of SamplesMinimum H-scoreMaximum H-scoreMean H-scoreMedian H-scoreNSCLC3034230117.4117OvCa2424240151151

Citation Format: Pamela M. Whalen, Amy Jackson-Fisher, Pamela Vizcarra, Cory Painter, Steven Pirie-Shepherd, Diane R. Fernandez, Joseph Lee, Eric L. Powell. Development of a novel PTK7 immunohistochemistry (IHC) assay and analytical validation in epithelial malignancies of lung, breast and ovary [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 4925.
Abstract 493: The landscape of adenosine signaling in cancer
Authors: Sidders Ben , Zhang Pei , Goodwin Kelly , O'Connor Greg , Borodovsky Alexandra , Dry Jonathan , Woessner Richard , Sachsenmeier Kris
Published: August 2019

Adenosine is a key suppressor of immune cell function. CD73 and other nucleotidases/phosphatases dephosphorylate and convert extracellular AMP into adenosine, which binds the A2A adenosine receptor (A2aR). There are several agents in early clinical trials targeting components of the adenosine pathway including A2AR and CD73. The identification of cancers with a significant adenosine drive is therefore critical to our understanding of the potential for these molecules. However, it is challenging to measure tumour adenosine levels on a pan-cancer scale. Here we describe the generation & validation of a gene expression signature that we use to characterise the pan-cancer landscape of adenosine signalling and its role within the tumour microenvironment.

Natural Language Processing (NLP) was used to define a network of regulatory interactions between the A2aR receptor and downstream entities. 14 genes were identified as being positively regulated by A2aR and robustly expressed in tumours and so constitute the signature, which we demonstrate accurately recapitulates adenosine drive in mouse syngeneic models. We therefore applied the signature to a pan-cancer analysis of tumour transcriptomes from TCGA and found a significant negative impact on both overall survival (HR = 0.6, Cox PH p=0) and progression free survival (HR = 0.77, Cox PH p=0.0000006) in adenosine high tumours. An adenosine related survival deficit was also seen in CD8+ T-cell infiltrated tumours (OS HR = 0.47, Cox PH p=0; PFS HR = 0.65, Cox PH p=0.0000002). Studying adenosine signalling in the context of tumour genetics identified highly significant associations with TGFB super-family members including TGFBR2 (p=0.0000000013). These data support the adenosine pathway as a key mediator of a successful anti-tumour immune response, demonstrate the prognostic potential of the gene expression signature and could inform patient selection strategies for adenosine pathway modulators.

Citation Format: Ben Sidders, Pei Zhang, Kelly Goodwin, Greg O'Connor, Alexandra Borodovsky, Jonathan Dry, Richard Woessner, Kris Sachsenmeier. The landscape of adenosine signaling in 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 493.
Abstract 4939: Alteration of TGFB signaling pathway predicts worse prognosis in pancreatic ductal adenocarcinoma
Authors: Li Mao , Li Ang , Zhao Xiaochen , Hou Shengzhong , Lu Shan , Mou Yu , Liu Xubao , Cao Dan , Cai Shangli , Tian Bole
Published: August 2019

Background: For pancreatic ductal adenocarcinoma (PDAC), a reliable and practical prognostic biomarker is still in unmet needs. Transforming growth factor beta (TGFB) signaling pathway is one of the core pathways involved in tumor initiation and progression via canonical and non-canonical pathway. The prognostic value of TGFB pathway genes as a functionally related group in PDAC is rarely studied.Methods: 72 PDAC patients who underwent surgery between November 30, 2015 and September 13, 2017 in West China Hospital, Sichuan University were identified and included in this study. Whole-exome sequencing (WES) or targeted next-generation sequencing was performed with tumor tissue from primary tumors. Clinical data was retrospectively collected. Clinicopathologic characteristics and overall survival were analyzed.Results: Genetic alterations were detected in 70 patients (97.2%). While 1 patient (1.4%) had only one genetic alteration, 28 patients (38.9%) had 2-4 genetic alterations and 43 patients (59.7%) had ≥5 genetic alterations. Overall, 25 patients (34.7%) with alteration of TGFB pathway and 47 patients (65.3%) with wild type of TGFB pathway were identified as TGFBm+ group and TGFBm- group respectively. TGFBm+ group had worse overall survival (HR, 1.875, 95% CI: 1.06-4.72; P = 0.04), especially in patients who had radical surgery (HR, 3.11, 95% CI: 1.11-20.3; P = 0.04). While in the subgroup of patients who underwent palliative surgery, OS was not statistically different (HR, 0.93, 95% CI: 0.42-2.04; P = 0.85). Subgroup analysis showed that overall survival hazard ratio favored participants with wild type of TGFB pathway in patients with ECOG performance score >1, abnormal CA199, without metastasis, stage I to II and patients who accepted radical surgery.Conclusion: Alteration of TGFB pathway genes is prevalent in PDAC. Inferior prognosis was observed in PDACs with mutations of TGFB pathway. TGFB signaling might play different role in different tumor stages, which is consistent with previous report about paradoxical effect of TGFB pathway. Genomic sequencing could help screen out patients at risk after surgery and adjuvant therapy might benefit this subgroup of PDACs.

Note: This abstract was not presented at the meeting.

Citation Format: Mao Li, Ang Li, Xiaochen Zhao, Shengzhong Hou, Shan Lu, Yu Mou, Xubao Liu, Dan Cao, Shangli Cai, Bole Tian. Alteration of TGFB signaling pathway predicts worse prognosis in pancreatic ductal adenocarcinoma [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 4939.
Abstract 4941: TTF1 negative in non small cell lung cancer adenocarcinoma: A prognostic factor
Authors: Cordoba Juan F. , Morales Serafin , Veas Joel , Rodriguez Alvaro , Reguart Noemi , Salud Antonieta
Published: August 2019

BACKGROUND: Patients with thyroid transcription factor 1 (TTF1) negative lung adenocarcinoma (ADC) have been reported to have a worse prognosis and to lack epidermal growth factor receptor (EGFR) mutations. This study describes a series of sample tumors from patients with clinically confirmed lung cancer.

METHODS: In all lung cancer patients diagnosed in the University Hospital of Lleida from January 2011 to December 2016, a real world data study of TTF1-negative ADC was performed, using TTF1 clone 8G7G3/1(DAKO®). Each patient’s clinical history, pathology specimens and molecular results were noted. Two hundred and thirty one patients with TTF1 positive lung ADC formed the control group.

RESULTS: Twenty seven (27) patients were identified with ADC TTF1 negative (74% males). The media age was 66 years. The smoking history was as follows: 55,5% former, 29,6% ex smokers and 14,8% never smokers. The clinical stages were as follows: stage I or II (n = 3 [11%]), stage III (n = 4 [14,8%]) and stage IV (n = 20 [74%]). Patients’ mean survival was 7,6 vs 24,4 months in ADC TTF1 negative patients (P= 0,00001). Three hundred and thirty four (334) patients with ADC were found, TTF1 results were available in 258 (77,2%) cases, of them twenty-seven cases (10,4%) had TTF1 negative; when they were compared with the control group, TTF1 negative patients had overall shorter survival (P= 0,00001), regardless of whether patients were treated with radical or palliative intent: 34,4 vs 10, months P=0,00001 (radical treatment) and 14,6 vs 6,7 months P= 0,007 (palliative intent). EGFR mutations were less frequent (P= 0,046) in TTF-negative tumors 5,5% vs 25,3%.

CONCLUSIONS: Patients with TTF1 negative NSCLC ADC, have worse overall survival regardless of treatment intention and a lower frequency of EGFR mutations.

KEYWORDS: lung adenocarcinoma (ADC), non small cell lung cancer (NSCLC), epidermal growth factor receptor (EGFR), prognosis in thyroid transcription factor 1 negative (TTF1) adenocarcinoma.

Citation Format: Juan F. Cordoba, Serafin Morales, Joel Veas, Alvaro Rodriguez, Noemi Reguart, Antonieta Salud. TTF1 negative in non small cell lung cancer adenocarcinoma: A prognostic factor [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 4941.
Abstract 4947: Plucked anagen scalp hair: A comparison between different analytical techniques to establish reproducible drug induced transcriptional changes and provide pharmacodynamic biomarker and target engagement information from oncology patients
Authors: Mefo Tim , Harrison Elliott , Murdoch Alan , Reed Benjamin
Published: August 2019

Plucked anagen scalp hair is an ideal surrogate tissue to monitor drug induced transcriptional changes. High vascularisation of the hair follicle, ease of sampling and high degree of similarity of expression in hair of pathways dysregulated in cancers, make the cellular bulb of plucked human scalp hair an excellent surrogate tissue for the non-invasive monitoring of pharmacodynamic (PD) and mechanism of action (MOA) effects in clinical trials. Using our ex vivo plucked hair culture platform, plucked scalp hair bulbs from several healthy donors were exposed to BEZ235, a dual pan-class PI3K and mTOR inhibitor at different concentrations and durations. Total RNA was isolated from the cellular bulb of anagen hair post-culture and samples were assessed by microarray and RNA-seq analysis to assess dose dependent global transcriptional alterations and develop a gene expression signature indicative of BEZ235 exposure. Two different microarrays, the Affymetrix Clariom D and U133 plus 2.0 arrays, were directly compared to RNA-seq data obtained from the same ex vivo culture samples. Genetic signatures indicative of exposure of plucked scalp hair to BEZ235 were established using all three analytical techniques. The transcriptional readouts from the plucked scalp hair samples revealed strong correlations (0.97-0.99) in the genes expressed in anagen scalp hair between donors. In addition, global gene expression data indicated that assessing pooled scalp hairs was sufficient to detect significant differential expression (p<0.05 & 1.5 fold change) underpinning the low variance in the majority of target genes in plucked hair as a biomarker platform. Gene signatures for compounds targeting the PI3K/mTOR pathway that were established using the ex vivo plucked scalp hair platform were subsequently used to support clinical trials and scalp hair taken from patients receiving treatment showed an excellent post-exposure target signature modulation. In summary, the results of the ex vivo plucked assessment generated a dose dependent and biologically relevant transcriptional signature following exposure to BEZ235. Plucked anagen scalp hair is therefore an ideal non-invasive surrogate tissue to obtain drug-induced pharmacodynamic biomarker and target engagement information from oncology patients.

Citation Format: Tim Mefo, Elliott Harrison, Alan Murdoch, Benjamin Reed. Plucked anagen scalp hair: A comparison between different analytical techniques to establish reproducible drug induced transcriptional changes and provide pharmacodynamic biomarker and target engagement information from oncology patients [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 4947.
Abstract 4963: A novel intestinal microbiome-derived peptide modulates immune cell activity and the tumor microenvironment
Authors: Kiefel Helena , Haria Dhwani , Katlinskaya Yuliya , Ravichandar Divya , McLaughlin Lily , Jain Sunit , Weinmaier Thomas , Iwai Shoko , DeSantis Todd , Takeuchi Toshi , Dabbagh Karim , Graham Kareem
Published: August 2019

The composition of the gut microbiota affects cancer development, progression, and response to therapy. A number of commensal bacteria, including Bifidobacterium, have been associated with increased response to immune checkpoint inhibitors in mouse tumor models and in cancer patients.

We hypothesized that secreted peptides or proteins are driving the Bifidobacterium-mediated effects. Using our unique bioinformatic-driven discovery platform we nominated putatively secreted Bifidobacterium-derived peptides for evaluation in immune cell effector assays. We demonstrate that several of the peptides induce secretion of pro-inflammatory cytokines (e.g., IL-6, TNF) by in vitro-generated mouse and human dendritic cells, as well as effector cytokine secretion (e.g., IFNγ, IL-2) by mouse splenic T lymphocytes in vitro.

To investigate the function of Bifidobacterium-derived peptides in the context of the tumor microenvironment we injected the candidate peptide SG-A directly into the tumor of CT26 tumor-bearing mice and analyzed the immune phenotype using the Nanostring PanCancer Immune Profiling panel and flow cytometry. Both Nanostring as well as flow cytometry analysis showed an increase in CD45+ tumor-infiltrating lymphocytes in SG-A treated tumors. SG-A peptide also induced the upregulation of dendritic cell function genes (CD40, CD83, and CD86) and multiple effector cytokines and chemokines. Re-stimulation of tumor-draining lymph node cells with AH1 peptide (a CT26 tumor-derived antigen) also increased induction of IFNγ in SG-A treated animals (vs. vehicle-treated controls).

Collectively, our results demonstrate the utility of the Second Genome discovery platform for leveraging microbiome science to identify novel immunoregulatory factors. This platform offers a promising approach to identify agents with potential for use as therapeutics in cancer immunotherapy.

Citation Format: Helena Kiefel, Dhwani Haria, Yuliya Katlinskaya, Divya Ravichandar, Lily McLaughlin, Sunit Jain, Thomas Weinmaier, Shoko Iwai, Todd DeSantis, Toshi Takeuchi, Karim Dabbagh, Kareem Graham. A novel intestinal microbiome-derived peptide modulates immune cell activity and the tumor microenvironment [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 4963.
Abstract 4966: Characteristics of immune repertoires in early lung cancer patients
Authors: Wang Jin , Hu Xihao , Zhang Peng , Liu Xiaole Shirley
Published: August 2019

Lung cancer is the leading cause of cancer death in the world, and early lung cancer detection is vital for improving the overall survival. Besides x-ray and CT scan, few genetic tests have been widely used for detecting early lung tumors. We hypothesize that early lung cancer patients have some unique features in the immune repertoires which can be utilized for early cancer detection and profiling disease development. In this study, we performed RNA-seq for paired tumor and tumor-adjacent normal samples from 59 early lung adenocarcinoma patients. We found that early lung tumors have more cytokine activities, higher B cell and CD4+ T cell infiltration, and lower CD8+ T cell infiltration compared to normal lung tissues. We assembled B cell receptors from RNA-seq reads and found more B cell clonal expansion and somatic hypermutations in the tumor than in tumor-adjacent normal tissues, similar to the B cell repertoires in late-stage lung cancer patients. In both early- and late-stage lung tumors, we identified a few cytokines that positively correlated with more IgA isotypes and associated with patient survival. To gain more insights on the B cell binding specificity, we performed motif enrichment analysis for paired tumor and normal samples and validated some tumor-enriched receptor motifs in matched patient blood samples, which implies a new way of early cancer detection. More interesting, we assembled microbial transcripts from RNA-seq data and identified some known lung disease-related species in both tumor and normal samples. The complex interactions between cancer cells, B cells, and microbiota in the early lung tumor development are waiting to be elucidated.

Citation Format: Jin Wang, Xihao Hu, Peng Zhang, Xiaole Shirley Liu. Characteristics of immune repertoires in early lung cancer patients [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 4966.
Abstract 4974: Germline prognostic SNPs and tumor-immunity-specific expression QTL (tis-eQTL) in cancer
Authors: Yu Xiaoqing , Wang Xuefeng
Published: August 2019

Introduction: Immunotherapy has produced promising results in treating cancers. Recent studies that aim to predict which patients can benefit from an immune checkpoint blocker have been largely focused on tumor molecular profiling such as tumor mutation burden and T cell infiltrations. However, whether the immunity landscape in tumor can be favorably or adversely affected by polymorphisms carried in the germline remains unknown and understudied.

Methods: Here we conduct the largest investigation of tumor-immunity-specific expression QTL to date, or tis-eQTL, to systematically identify germline genetic variants that affect immune landscape in tumor. We develop a reliable statistical method for tis-eQTL mapping using RNA-seq data from heterogeneous tumor samples. We analyze genomic data from 10,380 cancer patients from 33 cancer types to reveal interactions between genetic variants (derived from germline SNP array and WES data) and immune- phenotypes derived from tumor RNAseq data. These phenotypes include immune gene expression (such as CD3E, GZMA, CXCR3 and PRF1), T-cell receptor (TCR) clonality, and antigen presenting scores (APM).

Results: We observed that stratifying patients by the prioritized pathogenic germline polymorphisms exposed distinct tumor immune landscape, implicating new prognostic factors or risk genes of cancer. We further prioritize SNPs that fulfill two criteria: (a) with differential overall survival (b) have a differential expression pattern in immunogenic signatures. In support of these findings, we show that the top-ranked SNPs are associated with treatment responses to anti-PD-1 therapy in melanoma and non-small cell lung cancer. This study creates a unique and pioneering resource of prognostic germline variants with the potential to modulate immune therapy response and cancer risk, opening new avenues for developing next generation therapeutics and for personalizing risk assessment.

Note: This abstract was not presented at the meeting.

Citation Format: Xiaoqing Yu, Xuefeng Wang. Germline prognostic SNPs and tumor-immunity-specific expression QTL (tis-eQTL) in 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 4974.
Abstract 4975: Using humanized B-hSIRPa/hCD47 mouse model to evaluate the efficacy and toxicity of CD47 and SIRPa antibodies
Authors: Li Yanan , Guo Chaoshe , Xiang Jie , Lin Qingcong
Published: August 2019

Cancer immunotherapy is one of the most promising research areas in the field of cancer therapy. Along the IO drug development process, production of mouse models for in vivo efficacy evaluation has always been a rate-limiting step. Therefore, we generated a novel humanized knock-in mouse to evaluate the efficacy of human IO antibodies.

CD47 is a transmembrane protein expressed in many human tissues. SIRPa, which is expressed on phagocytic cells, was identified as the receptor of CD47. Engagement of SIRPa by CD47 serves as “do not eat me” signal, inhibiting the phagocytic activity of macrophages. Antibodies interfering with the CD47-SIRPa interaction block inhibition and promote tumor destruction. While CD47 is a potential target for antibody development, side effects associated with CD47 blockade have emerged as a major concern, especially anemia.

To evaluate the efficacy and toxicity of CD47 antibody, we generated a double humanized B-hSIRPa/hCD47 mouse. In this model, exon2 in mouse SIRPa and CD47 were replaced with the corresponding human sequences. In homozygous mice, mouse SIRPa and CD47 was absent and only human protein expression was detected, B-hSIRPa/hCD47 mice paired with genetically modified MC38-hCD47 cancer cells were used to screen CD47 antibodies by balancing their efficacy and toxicity. Antibodies with less toxicity exhibited minimal weight loss, minimal effects on blood cell and platelet production, and reduced ALT/AST elevation in mice. Thus, we demonstrated that these humanized mice could expedite and improve the development of safer CD47 antibodies.

Humanized B-hSIRPa/hCD47 mice can also be used to evaluate the efficacy of antibodies targeting SIRPa. We also generated triple humanized B-hPD-1/hSIRPa/hCD47 and B-hPD-L1/hSIRPa/hCD47 mice that would be useful to develop combination therapies of PD-1/PD-L1 and SIRPa/CD47 antibodies. Humanized B-hSIRPa/hCD47 mice are a promising in vivo efficacy model for the development of CD47 and SIRPa antibodies that can be advanced to human clinical trials.

Citation Format: Yanan Li, Chaoshe Guo, Jie Xiang, Qingcong Lin. Using humanized B-hSIRPa/hCD47 mouse model to evaluate the efficacy and toxicity of CD47 and SIRPa antibodies [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 4975.
Abstract 498: Integrating bulk and spatial profiling technologies for the discovery of RNA and protein biomarkers in muscle invasive bladder cancer
Authors: Zhang Zhaojie , Das Subhasree , Korpal Manav , Kumar Pavan , Gong Jingjing , Liang Yan , Hoang Margaret , Beecham Joseph , Rimkunas Victoria
Published: August 2019

Muscle-invasive bladder cancer (MIBC) is an aggressive disease with limited therapeutic options. Although immunotherapies are approved for MIBC, the majority of patients fail to respond, suggesting the existence of complementary immune evasion mechanisms underscoring the need for comprehensive immune profiling of patient tumor samples. The PPARγ pathway constitutes a tumor-intrinsic mechanism underlying immune evasion in MIBC. Immune cell-infiltration is controlled by activated PPARγ, which in turn inhibits expression/secretion of inflammatory cytokines. Clinical data sets indicate that PPARγHigh impairs CD8+ T-cell infiltration and confers partial resistance to immunotherapies.

Here, we report a comprehensive biomarker study aimed at identifying a single, highly multiplexed platform for profiling the tumor microenvironment in clinical-type specimens as formalin-fixed, paraffin-embedded (FFPE) samples. A cohort of PPARγHigh and PPARγLow muscle invasive bladder cancer samples were used for the technology assessments. Gene expression patterns were measured using a combination of research use only genomics and spatial genomic technologies to determine concordance including RNA-seq, spatial in situ hybridization (GeoMx™ Digital Spatial Profiler) and IO360. We determined that multiple RNA and protein-based technologies were able to accurately characterize the immune phenotypes of PPARγHigh tumors. However, the GeoMx™ technology was ideal for precious samples since it digitally quantifies target abundance while maintaining spatial resolution in only two FFPE slides.

GeoMxTM DSP technology is for Research Use Only and not for use in diagnostic procedures.

Citation Format: Zhaojie Zhang, Subhasree Das, Manav Korpal, Pavan Kumar, Jingjing Gong, Yan Liang, Margaret Hoang, Joseph Beecham, Victoria Rimkunas. Integrating bulk and spatial profiling technologies for the discovery of RNA and protein biomarkers in muscle invasive bladder 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 498.
Abstract 4988: CD 137 agonists as an adjunct to immune checkpoint inhibitors to overcome resistance in melanoma
Authors: Danturti Sreedevi , Mayer Lena , Victor Christina Twyman Saint
Published: August 2019

Background: Melanoma is currently the fifth most common cancer in men and sixth most common in women with an estimate of 9,320 deaths from the disease in the United States. Anti-PD1, an immune checkpoint inhibitor, is FDA approved to treat metastatic melanoma with durable responses observed in 53% of patients. Studies suggest that combining therapies (e.g. immune checkpoint inhibitors, radiation therapy, and/or agonist antibodies) can be more effective than monotherapy. CD137 is expressed on a variety of cell populations including T cells, dendritic cells (DCs) and macrophages. CD137 agonists have proved effective in multiple murine tumor models and have had modest responses in early clinical trials. We hypothesize that CD137 agonists can augment the anti-PD1 anti-tumor immune response resulting in improved overall survival and decreased tumor burden in melanoma.

Methods: 499 melanoma cells, which were derived from B16-F10, are implanted on both flanks of C57BL6 mice. The mice are treated with anti-CD137 and/or anti-PD1 on day 10, 13 and 16 post-implantation. The anti-tumor response is assessed by both overall survival and tumor burden. Tumors are harvested on day 19 for protein, transcriptomic and immune profiling.

Results: Monotherapy with either anti-CD137 agonists or anti-PD1 antagonists increases the overall survival by 8 days; whereas, combination therapy results in an increase of 25 days. To investigate the mechanisms of response and resistance to combination therapy, tumors were harvested and assessed via a proteomic array that identified 36 proteins that were upregulated in the non-responders. Transcriptomic profiling of these tumors using Nanostring IO 360 corroborated these findings and identified both the myeloid the T cell immune populations are upregulated in responders.

Conclusions: These results suggest a dual role for both the innate and adaptive immune response in treating melanoma. By understanding the mechanism of response and further characterization we can further best apply the combination therapy to overcome resistance after immunotherapy.

Note: This abstract was not presented at the meeting.

Citation Format: Sreedevi Danturti, Lena Mayer, Christina Twyman Saint Victor. CD 137 agonists as an adjunct to immune checkpoint inhibitors to overcome resistance in melanoma [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 4988.
Abstract 5020: Therapeutic potential of an immune modulatory CCL22-based vaccine
Authors: Lecoq Inés , Kopp Katharina L. , Christensen Rikke , Pedersen Ayako W. , Martinenaite Evelina , Zocca Mai-Britt , Andersen Mads H.
Published: August 2019

CCL22 is a macrophage-derived chemokine that exerts immunosuppressive functions by the recruitment of regulatory T cells (Tregs) through the CCL22/CCR4 axis. It is expressed by cells in the tumor microenvironment (TME) in many different cancer tissues including breast, lung, and pancreatic cancer and is thought to promote the suppression of anti-cancer immunity. Recently, we described that CCL22-specific T-cells from cancer patients can kill CCL22-expressing tumor cells, and activation of CCL22-specific T cells may directly influence the CCL22 concentration in the TME, suggesting that CCL22 may be an interesting target for an immune modulatory vaccine.

Thus, in the present study we have targeted CCL22-expressing cells by activation of CCL22-specific effector T cells by peptide vaccination in tumor models. We present here that CCL22-targeting peptide vaccination leads to expansion of predominantly CD8+, IFNγ-secreting CCL22-specific T cells in both BALB/c and C57BL/6 mice. Furthermore, CCL22 vaccination results in delayed tumor growth and enhanced survival in tumor harboring hosts in CT26 and Pan02 models. Preliminary assessment of gene expression in the tumors reveals that CCL22 vaccination leads to a reduction of CCL22 expression, alongside other immune suppressive molecules, such as IDO1 in the TME. Characterization of vaccine-mediated changes of cellular composition of the TME is currently ongoing.

In conclusion, our findings provide a rationale for a novel cancer immunotherapy to target the immune suppressive TME.

Citation Format: Inés Lecoq, Katharina L. Kopp, Rikke Christensen, Ayako W. Pedersen, Evelina Martinenaite, Mai-Britt Zocca, Mads H. Andersen. Therapeutic potential of an immune modulatory CCL22-based vaccine [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 5020.
Abstract 5030: Microtubule destabilization by plinabulin generates anti-tumor immune response through repolarization of intratumoral macrophages
Authors: Bejestani Elham Pishali , Lloyd G. Kenneth , Buchi Melanie , Ritschard Reto , Herzig Petra , Mohanlal Ramon , Huang Lan , Tonra James R. , Zippelius Alfred , Kashyap Abhishek
Published: August 2019

The M2 phenotype is dominant in tumor associated macrophages (TAMs) and correlates with poor prognosis. As M1 polarized macrophages possess anti-tumor functions, in vivo reprogramming of macrophages is a promising strategy for cancer treatment. We recently demonstrated that microtubule destabilizing chemotherapeutic agents such as plinabulin induce potent dendritic cell maturation and thereby augment anti-tumor immune responses; yet the effects of plinabulin on macrophage polarization remain unknown. Here we demonstrate that plinabulin, which is currently being investigated for both its anti-tumor effect and prevention of chemotherapy-induced neutropenia in phase II/III clinical trials,induces M1 polarization of ex vivo cultured human monocyte-derived M2 macrophages. This was characterized by a reduction in the cell surface expression of M2 markers CD163 and CD206 and an increase in the expression of co-stimulatory M1 markers CD80 and CD86. Phenotypically, plinabulin-polarized human M1 macrophages secreted increasing levels of iNOS and inflammatory cytokines IL-1β, IL-6 and IL-12 while IL-10 and IL-4 secretion decreased. This suggests a functional skewing from immunosuppressive M2 to pro-inflammatory M1 macrophages upon exposure to plinabulin. Similarly, plinabulin induced the M1 polarization of murine FACS sorted F4/80+ intratumoral M2-TAMs, characterized by increased CD80 and decreased CD206 expression. Moreover, systemic treatment (i.p.) of tumor bearing mice with plinabulin led to a significant reduction in the number of tumor-infiltrating TAMs with a concomitant M1 polarization of the remaining TAMs. Indeed, plinabulin treatment of mice bearing subcutaneous MC38 colon cancer or orthotopic EMT6 breast cancer tumors significantly delayed tumor growth. This efficacy of plinabulin remained unaltered in Rag2-/- mice lacking T cells, suggesting that macrophages are required for its anti-tumor activity. These results identify targeting of TAMs by plinabulin as a promising therapeutic strategy. Testing of plinabulin in combination with other macrophage targeting drugs are currently being explored to investigate the dual effect in reprogramming the tumor immune microenvironment.

Citation Format: Elham Pishali Bejestani, G. Kenneth Lloyd, Melanie Buchi, Reto Ritschard, Petra Herzig, Ramon Mohanlal, Lan Huang, James R. Tonra, Alfred Zippelius, Abhishek Kashyap. Microtubule destabilization by plinabulin generates anti-tumor immune response through repolarization of intratumoral macrophages [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 5030.
Abstract 5042: Folate and other B-vitamins and the risk of breast cancer in younger women
Authors: Houghton Serena C. , Eliassen A Heather , Zhang Shumin , Selhub Jacob , Rosner Bernard A. , Willett Walter C. , Hankinson Susan E.
Published: August 2019

Purpose: Prior epidemiologic cohort studies have generally found no associations of folate and other B-vitamins with breast cancer overall, but differences by menopause status have been suggested. However, prior studies have been limited by small numbers to examine the relationship between different plasma measures and related polymorphisms and breast cancer risk among younger women. Therefore, we examined the association of plasma B-vitamins and metabolites, and related genetic variants, with risk of breast cancer among predominantly premenopausal women.

Methods: We conducted a nested case-control study within the Nurses’ Health Study II. From blood samples collected in 1996-1999 and follow-up through 2007, plasma measures were available for 610 cases and 1,207 controls. Unconditional multivariable logistic regression was used to estimate relative risks (RR) of breast cancer and 95% confidence intervals (CIs). We examined whether associations varied by methylenetetrahydrofolate reductase (MTHFR) and dihydrofolate reductase polymorphisms, breast cancer risk factors, or tumor characteristics.

Results: Plasma vitamin B12 was associated with a 71% increase in risk of breast cancer comparing the highest versus lowest quintile (95%CI=1.22-2.39, p-trend=0.02). Plasma folate (comparable RR=1.20, 95%CI=0.85-1.68), pyridoxal 5’-phosphate (RR=1.19, 95%CI=0.85-1.65), cysteine (RR=1.15, 95%CI=0.81-1.63), homocysteine (RR=0.90, 95%CI=0.65-1.25), and cysteinylglycine (RR=0.92, 95%CI=0.65-1.31) were not associated with overall breast cancer risk. Folate was significantly positively associated with invasive (RR highest tertile vs. lowest=1.48, 95%CI=1.09-2.01) and estrogen receptor-positive/progesterone receptor-positive breast cancer (RR T3 vs. T1= 1.59, 95%CI= 1.12-2.27) and the invasive association was suggestively stronger for bloods collected post-fortification (p-heterogeneity=0.19). Several nutrient/breast cancer associations varied across subgroups defined by age, smoking, alcohol, multivitamin use and MTHFR status (p-interaction<0.05).

Conclusions: Overall, plasma B-vitamins and metabolites were not consistently associated with lower breast cancer risk. For increasing plasma folate and B12 levels higher risk of invasive breast cancer was observed. Additionally, there may be associations in subgroups defined by related genetic variants, breast cancer risk factors, and tumor factors. Future studies in younger women are needed to confirm findings.

Citation Format: Serena C. Houghton, A Heather Eliassen, Shumin Zhang, Jacob Selhub, Bernard A. Rosner, Walter C. Willett, Susan E. Hankinson. Folate and other B-vitamins and the risk of breast cancer in younger women [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 5042.
Abstract 5048: Determination of haptoglobin, hemoglobin genotypes and malaria incidence in Nigerian breast cancer patients
Authors: Dokunmu Titilope M. , Obi Patience , Rotimi Oluwakemi A. , Fatiregun Omolara A. , Agodirin Sulaiman O. , Rotimi Solomon O.
Published: August 2019

Breast cancer is the second leading cause of cancer morbidity and mortality globally. Cancer chemotherapy commonly result in hemolysis, which impacts patient overall health. There is a need to determine genetic factors associated with hemolysis in breast cancer patients. Haptoglobin (Hp), a polymorphic protein plays important role in hemoglobin clearance and disease predisposition, but has been reported to have no prognostic factor in breast cancer. However, understanding selection pressure that drives certain gene mutations in specific populations and how it confers protection or susceptibility to diseases is crucial. In Nigeria, breast cancer, malaria infection and sickle cell disease are prevalent and associated with hemolysis, but little is known of their association in breast cancer patients. This study aims to determine relationship between haptoglobin, hemoglobin genotypes and submicroscopic malaria co-morbidity in clinically diagnosed breast cancer and healthy Nigerian women. DNA was extracted from blood using standard methods. Haptoglobin 2 and hemoglobin genotypes were detected by RFLP-PCR, while Plasmodium falciparum infection was detected by primer specific amplification of plasmodium cytochrome oxidase III gene (cox III) in 75 clinically diagnosed breast cancer (BC) and 287 healthy women (control; HC). Proportions were determined and compared in the two groups and test of association was carried out with significance level set at P <0.05. In BC groups, 3 (4.1%) of 72 Hp 2-2 phenotypes was detected compared to a significantly higher occurrence of 48 (16.7%) of 287 in HC group (p <0.05). Conversely, malaria infection was detected in 68 (94.4%) BC versus 255 (88.9%) in HC group. A similar proportion had Hp deletions (2 in BC and 8 in HC group). There was a low prevalence of hemoglobin S genotype in the entire population and relative risk for Hp 2-2 polymorphism in hemoglobin genotypes was not significantly different. In conclusion, this study reports in breast cancer and healthy women an inverse correlation of haptoglobin (Hp2-2) genotype with malaria incidence in southwest Nigeria. The results imply a possible protection against hemolysis and can play significant role in determining choice of cancer therapy for good patient treatment outcomes.

Citation Format: Titilope M. Dokunmu, Patience Obi, Oluwakemi A. Rotimi, Omolara A. Fatiregun, Sulaiman O. Agodirin, Solomon O. Rotimi. Determination of haptoglobin, hemoglobin genotypes and malaria incidence in Nigerian breast cancer patients [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 5048.
Abstract 5059: Early life risk factors and childhood cancer risk
Authors: Stacy Shaina L. , Buchanich Jeanine M. , Ma Zhen-qiang , Mair Christina , Robertson Linda , Sharma Ravi K. , Talbott Evelyn O. , Yuan Jian-Min
Published: August 2019

Prenatal exposures, including maternal, delivery, and newborn characteristics, may affect childhood cancer risk. Our objective was to examine whether such birth certificate-derived factors are associated with increased risk of subsequent cancer development, including leukemia and central nervous system (CNS) tumors, in children ages zero to 13. Pennsylvania state birth and cancer registry files obtained from the PA Department of Health were linked to create a cohort of children born from 2003 to 2015 who developed cancer from 2003 to 2016. After exclusions, 1,834,796 infants were included with a total of 13,785,309 person-years of observation. Among these, there were 2,355 children diagnosed with any cancer, 748 with leukemia, and 545 with CNS tumors. Older maternal age increased the hazard of developing any childhood cancer, leukemia, and CNS tumors. Children born to severely obese mothers (body mass index ≥40 kg/m2) were at over 50% higher hazard of leukemia (P=0.01) after adjustment for age and other significant risk factors; however, maternal obesity did not significantly increase the risk of childhood CNS tumors. Among the infant characteristics, having a fetal growth ratio 30% higher than expected was significantly associated with increased hazard of any childhood cancer, leukemia, or CNS tumors (hazard ratios 2.19, 1.79, and 1.77, respectively). Other fetal and newborn factors associated with higher risk of total childhood cancer included fetal intolerance of labor, lower gestational age, and having a low Apgar score at birth. This analysis demonstrated significant and independent impacts of maternal obesity and fetal growth rate on childhood cancer development, particularly leukemia, after accounting for multiple risk factors. If confirmed, weight control during pregnancy would be an effective means of childhood cancer prevention.

Citation Format: Shaina L. Stacy, Jeanine M. Buchanich, Zhen-qiang Ma, Christina Mair, Linda Robertson, Ravi K. Sharma, Evelyn O. Talbott, Jian-Min Yuan. Early life risk factors and childhood cancer risk [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 5059.
Abstract 5064: Sirt6 is a novel tumor suppressor in human urothelial cancer and activated by methysticin
Authors: Fu DongJun , Yokoyama Noriko N. , Tippin Mattew , Zi Xiaolin
Published: August 2019

Sirtuin 6 (SIRT6) plays an important role in longevity, metabolism, DNA-repair, and inflammatory response. In human urothelial cancer, a high level of SIRT6 expression predicts a better survival for patients. We found that overexpression of SIRT6 in human muscle-invasive bladder cancer cell line J82 reduced cell migration and invasion, but didn’t reduce the in vitro cell growth. Kava is a traditional psychotropic beverage made from the rhizomes of Piper methysticum G. Forst. Previous epidemiological studies have shown an inverse relationship between cancer incidence and kava consumption in South Pacific Island Nations. We have shown that Methysticin, a major kavalactone in the Kava plant, inhibits the growth, migration and invasion of bladder cancer lines. However, these anti-cancer activities of Methysticin are attenuated in cells with low expression of SIRT6, expression of enzymatically mutant SIRT6 or SIRT6 knockout, which suggested that the anti-cancer activities of Methysticin are at least in part dependent on SIRT6 expression. Molecular modelling indicates that Methysticin docks into the site next to a loop near the acetylated peptide substrate binding site of SIRT6. The Cellular Thermal Shift Assay (CETSA, an in vivo assay) demonstrated that Methysticin significantly decreased heat-induced protein degradation of SIRT6. Treatment of J82 cells with Methysticin also resulted in a dose-dependent reduction of H3K18 acetylation. Taken together, these results suggest that Methysticin acts a novel SIRT6 activator for its anti-bladder activity. Further studies have demonstrated that Methysticin activates the TBK1/IRF3/STING pathway by increasing the protein expression of TBK1 and the phosphorylation of TBK1 and STING, which suggest that Methysticin may be able to enhance anti-tumor immunity. Therefore, our results support further investigation of Methylation for bladder cancer prevention.

Note: This abstract was not presented at the meeting.

Citation Format: DongJun Fu, Noriko N. Yokoyama, Mattew Tippin, Xiaolin Zi. Sirt6 is a novel tumor suppressor in human urothelial cancer and activated by methysticin [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 5064.