bluuebunny/crossref_metadata_2025 huggingface dataset:
Abstract 1410: Clinicopathological characteristics of lung cancer patients with concomitant idiopathic pulmonary fibrosis
Authors: Araki Kota , Shien Kazuhiko , Miyauchi Shunsaku , Miura Akihiro , Takahashi Yuta , Kurihara Eisuke , Ogoshi Yusuke , Namba Kei , Suzawa Ken , Yamamoto Hiromasa , Tomida Shuta , Soh Junichi , Sakaguchi Masakiyo , Toyooka Shinichi
Published: August 2019

Background: Idiopathic interstitial pneumonias (IIPs) are diffuse parenchymal lung disease of unknown etiology. Among IIPs, idiopathic pulmonary fibrosis (IPF) is the most frequent subtype. IPF is characterized by the accumulation of activated fibroblasts and extracellular matrix within the parenchyma. IPF is associated with pathologic and radiologic pattern of usual interstitial pneumonia (UIP). There are several possible underlying mechanisms linking lung cancer and IPF, however, many of them are still unknown. The purpose of this study is to reveal the clinicopathological characteristics of lung cancers patients with concomitant IPF.

Materials and Methods: The clinical and pathological data of lung cancer patients with IPF who underwent surgical resection of lung cancer between 2001 to 2016 at our institution were analyzed. We defined IPF as IIPs with UIP pattern, based on the chest CT scan and pathological findings. Mutations of EGFR (L858R point mutation and exon19 deletion), AKT1, BRAF, EGFR, KRAS, MEK1, NRAS, PIK3CA, and PTEN were analyzed.

Result: A total of 37 patients’ data of lung cancer with concomitant IPF was collected. Two patients had metachronous lung cancers. Thus, we analyzed 39 lung cancers from 37 patients. Based on the radiographic findings, 33 (84.2%) of the available 38 tumors arose from the fibrotic legion, which were mainly observed in peripheral region. The frequency of EGFR mutation was detected in 1 (2.6%) of 39 lung cancers. As control, we had EGFR mutation profile of 583 lung cancer without IPF. For this population, EGFR mutation was found in 164 (28.1%) of 583 cases. There was a significant difference in EGFR mutation between lung cancers with or without IPF (p = 0.0001). Among other examined genes in 16 tumors, one PIK3CA mutation was found.

Conclusion: The lung cancer with IPF mainly arouse from fibrotic lesion. The frequency of EGFR mutation was less common in lung cancers with IPF than those without IPF. Our result suggests that molecular profile of lung cancer with IPF is different from that without IPF.

Citation Format: Kota Araki, Kazuhiko Shien, Shunsaku Miyauchi, Akihiro Miura, Yuta Takahashi, Eisuke Kurihara, Yusuke Ogoshi, Kei Namba, Ken Suzawa, Hiromasa Yamamoto, Shuta Tomida, Junichi Soh, Masakiyo Sakaguchi, Shinichi Toyooka. Clinicopathological characteristics of lung cancer patients with concomitant idiopathic pulmonary fibrosis [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 1410.
Abstract 1420: Novel expansion of CD8+PD1+ spleen cells for therapeutic intent in pancreatic adenocarcinoma cancer
Authors: Cole Kathryn , Ly Quan P. , Cox Jesse L. , Hollingsworth Michael A. , Padussis James C. , Foster Jason M. , Vargas Luciano M. , Talmadge James E.
Published: August 2019

Pancreatic ductal adenocarcinoma (PDAC) is a devastating disease with poor prognosis and overall survival. In 2018 it is estimated that there will be 44,333 deaths making PDAC the 3rd cause of cancer related deaths. A distal pancreatomy for PDAC in the body/tail of the pancreas usually includes a splenectomy, with current practice to discard the spleen post-surgery. Spleens can provide a large number of lymphocytes, including CD8+ T-cells with potential to be utilized for adoptive cellular therapy. Our studies have focused on spleen from this patient population, which we posit (and show herein) provides a large number of CD8+PD1+ cells for expansion and the first clinical strategy to expand activated, tumor specific T-cells for subsequent infusion with therapeutic intent. In our studies we used Milteny magnetic bead isolation of CD8+PD1+ T-cells from the spleens of PDAC patients with disease in the distal pancreas. The purity of the cells was 98% with a yield of 79% providing 9x108 CD8+PD1+ cells from an idealized 160 gm spleen. CD8 expansion was undertaken with various growth factor cocktails incorporating combinations of three interleukins (IL); IL-7, IL-15, and IL-21 at varying concentrations. IL-7 supports T-cell survival and proliferation, IL-15 is a potent T-cell growth factor and promotes naïve and memory T-cell survival, and IL-21 limits T-cell exhaustion in response to chronic stimulation. The optimized cocktail incorporated IL-7, IL-15, and IL-21 at 40 ng/mL, which demonstrated maximum T-cell expansion, based on cellularity and cell viability. The predominant T-cell memory phenotype of the CD8+PD1+ cells following expansion was transitional memory (TTM) T-cells that increased from 35.6 ± 4.3 to 52 ± 3%. This memory T-cell phenotype has a high proliferative potential while retaining effector function. Following six days of culture with the three interleukins we observed a 10 fold increase in CD8+PD1+ cells such that 9x109 cells are obtained from the 160 gm spleen. These cells, based on ELISPOT analysis with autologous tumor lysate pulsed dendritic cells retained tumor specificity. In conclusion, CD8+PD1+ cells can be isolated from resected spleen, expanded with retention of tumor specificity and potentially used for adoptive cellular therapy.

Citation Format: Kathryn Cole, Quan P. Ly, Jesse L. Cox, Michael A. Hollingsworth, James C. Padussis, Jason M. Foster, Luciano M. Vargas, James E. Talmadge. Novel expansion of CD8+PD1+ spleen cells for therapeutic intent in pancreatic adenocarcinoma 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 1420.
Abstract 1422: Preclinical development of CD37CAR T-cell therapy for treatment of B-cell lymphoma
Authors: Dillard Pierre , Köksal Hakan , Josefsson Sarah , Maggadottir Solrun Melkorka , Pollmann Sylvie , Fåne Anne , Blaker Yngvild Nuvin , Beiske Klaus , Huse Kanutte , Kolstad Ane , Holte Harald , Kvalheim Gunnar , Smeland Erlend Bremertun , Myklebust June , Inderberg Else Marit , Wälchli Sebastien
Published: August 2019

T cells modified to express chimeric antigen receptor (CAR) targeting CD19 have produced remarkable clinical responses in patients with relapsed/refractory B-cell acute lymphoblastic leukemia (B-ALL). CD19CAR T-cell therapy has also demonstrated prominent effects in B-cell non-Hodgkin lymphoma (B-NHL) patients. However, a subset of patients who relapse after CD19CAR T-cell therapy have outgrowth of CD19-negative tumor cells. Hence, development of alternative CARs targeting other B-cell markers represents an unmet medical need for B-ALL and B-NHL. Here, we confirmed previous data by showing that B-NHL overall have high expression of CD37. A second generation CD37CAR was designed and its efficacy in T cells was compared to that of CD19CAR. In vitro assessment of cytotoxicity and T-cell function upon co-culture of the CAR T cells with different target B-cell lymphoma cell lines demonstrated comparable efficacy between the two CARs. In an aggressive B-cell lymphoma xenograft model, CD37CAR T cells were as potent as CD19CAR T cells in controlling tumor growth. In a second xenograft model, using U2932 lymphoma cells containing a CD19-negative subpopulation, CD37CAR T cells efficiently controlled tumors and cured the mice while CD19CAR T cells had limited effect. We further show that, unlike CD19CAR, CD37CAR was not sensitive to antigen masking. Finally, CD37CAR reactivity was restricted to B-lineage cells. Collectively, our results demonstrated that CD37CAR T cells effectively can eradicate B-cell lymphoma tumors also when CD19 antigen expression is lost, and support further clinical testing for patients with relapsed/refractory B-NHL.

Citation Format: Pierre Dillard, Hakan Köksal, Sarah Josefsson, Solrun Melkorka Maggadottir, Sylvie Pollmann, Anne Fåne, Yngvild Nuvin Blaker, Klaus Beiske, Kanutte Huse, Ane Kolstad, Harald Holte, Gunnar Kvalheim, Erlend Bremertun Smeland, June Myklebust, Else Marit Inderberg, Sebastien Wälchli. Preclinical development of CD37CAR T-cell therapy for treatment of B-cell lymphoma [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 1422.
Abstract 1431: In vivo CRISPR/Cas9 screen of adoptively transferred T cells reveals novel mediators of T cell exhaustion and immunosuppression
Authors: Park Jin K. , Canner David A. , Herbst Rebecca , Li Amy , Smith Olivia C. , Regev Aviv , Jacks Tyler E.
Published: August 2019

Adoptive cell therapy (ACT) is a form of cancer immunotherapy in which patients receive an infusion of a polyclonal population of autologous CD8+ T cells that can be genetically modified ex vivo to optimize their function against the tumor. Although clinical responses to ACT have been successful for a very limited number of indications (most notably pediatric leukemias), its efficacy in solid tumors is very limited, likely due to immunosuppressive mechanisms constraining efficacy. To date, there have not been systematic approaches to discover novel T cell-intrinsic mediators of exhaustion in the context of ACT. Here, we describe a systematic and unbiased approach to uncover novel regulators of exhaustion in adoptively transferred CD8+ T cells. We report the results of longitudinal RNA sequencing of adoptively transferred cells at various time points post-transfer, revealing the complex transcriptional changes that transferred cells undergo. In addition, we report the results of an in vivo CRISPR/Cas9-mediated genetic screen. Taken together, this integrated approach of longitudinal RNAseq and CRISPR/Cas9 screening has yielded mediators previously uncharacterized in T cells that can be targeted to drastically improve functionality, cytotoxicity, and persistence in the tumor microenvironment.

Note: This abstract was not presented at the meeting.

Citation Format: Jin K. Park, David A. Canner, Rebecca Herbst, Amy Li, Olivia C. Smith, Aviv Regev, Tyler E. Jacks. In vivo CRISPR/Cas9 screen of adoptively transferred T cells reveals novel mediators of T cell exhaustion and immunosuppression [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 1431.
Abstract 1432: Enhanced effector responses of regenerated CAR-T cells derived from genome edited iPSCs
Authors: Ueda Tatsuki , Iriguchi Shoichi , Kawai Yohei , Minagawa Atsutaka , Miyoshi Hiroyuki , Terakura Seitaro , Uemura Yasushi , Woltjen Knut , Kodama Yuzo , Seno Hiroshi , Hitoshi Yasumichi , Nakatsura Tetsuya , Tamada Koji , Kaneko Shin
Published: August 2019

Chimeric antigen receptor (CAR) is an artificial protein that provides HLA-independent antigen specificity to T cells. CAR-T therapy has shown remarkable clinical responses especially in hematologic malignancies. But this therapy requires cell preparation for each patient and it cause some limitations for applicability of CAR-T therapy. We have reported regeneration of T cells from iPSCs (Cell Stem Cell. 2013). Since this technology can provide unlimited number of T cells, CAR-T therapy using iPSCs is thought to broaden its applicability. To target solid tumors, it is important to avoid immunosuppressive factors from tumor microenvironment and to exert sufficient cytotoxicity. Tumor reactive T cells are known to fall into anergy state by continuous antigen stimulations. To overcome immunosuppression in tumor microenvironment, enhancement of TCR signaling by modification of genes related to TCR signal is a promising strategy. Recent studies revealed that the efficacy of CAR-iPS-T cells are not equivalent to primary CAR-T cells. To enhance the efficacy of CAR-iPS-T cells and to produce resistant CAR-T cells to immunosuppression, we focused on TCR signaling pathway. We found that antigen reactivity of CAR-iPS-T cells was insufficient compared with primary CAR-T cells. To overcome the weakness of TCR signal, we disrupted genes negatively related to TCR signal and successfully enhanced TCR signal. As a result, genome edited CAR-iPS-T cells could persist longer in vivo and displayed enhanced tumor suppressive function comparable with primary CAR-T cells. Genome edited iPSCs can be a unlimited cell source of enhanced CAR-T cells. These findings indicate that regenerated CAR-T cells derived from genome edited iPSCs would be a promising CAR-T therapy which would overcome immunosuppressive tumor microenvironment.

Citation Format: Tatsuki Ueda, Shoichi Iriguchi, Yohei Kawai, Atsutaka Minagawa, Hiroyuki Miyoshi, Seitaro Terakura, Yasushi Uemura, Knut Woltjen, Yuzo Kodama, Hiroshi Seno, Yasumichi Hitoshi, Tetsuya Nakatsura, Koji Tamada, Shin Kaneko. Enhanced effector responses of regenerated CAR-T cells derived from genome edited iPSCs [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 1432.
Abstract 1436: Development of a chimeric antigen receptor against glycosylated annexin A2 on ovarian tumors
Authors: Leong Leonard W. , Cua Simeon , Tan Heng Liang , Choo Andre B.
Published: August 2019

Chimeric antigen receptors (CARs) have been a significant focus of cancer therapeutic investigation in recent years, due to the remarkably effective responses observed in hematological cancers. However, the utility of CAR T cells has yet to be fully exhibited in solid tumours. One of the key challenges facing the treatment of solid tumors with CAR T cells is the dearth of viable antigen targets, which limits the expansion of this therapeutic space.

Here, we describe how the anti-glycan monoclonal antibody 2448, which target annexin A2 on a number of ovarian and breast cancer cell lines, has been successfully reformatted as a CAR T cell construct.

Messenger RNA (mRNA) encoding CAR(2448) was transfected into activated T cells, resulting in cytotoxicity and cytokine release upon co-incubation with target cells expressing annexin A2. While CAR(2448) variants with differing spacer chain lengths all mediated cytotoxicity against target cells compared to untransfected T cells, real-time target cell growth monitoring revealed that the long spacer CAR(2448) mediated higher levels of cytotoxicity, consistent with the membrane-proximal location of the annexin A2 target epitope.

This optimization of CAR spacer length in the in vitro setting, using mRNA transfected T cells and real-time target cell growth monitoring, suggests that several aspects of CAR construction can be optimized in vitro, thereby lowering the threshold for CAR T cell development.

These results suggest the potential for anti-glycan antibodies to be repurposed as CARs, and the development of CAR(2448) is currently being extended to lentiviral vectors for the purpose of developing long-term expressing CAR(2448) T cells, in anticipation of in vivo studies in murine models.

This project was funded by the Agency for Science, Technology, and Research (A*STAR), Singapore and the A*STAR Graduate Academy (A*GA).

Citation Format: Leonard W. Leong, Simeon Cua, Heng Liang Tan, Andre B. Choo. Development of a chimeric antigen receptor against glycosylated annexin A2 on ovarian tumors [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 1436.
Abstract 1437: Improving efficacy and accuracy of CAR-T cells for neuroblastoma
Authors: Moghimi Babak , Muthugounder Sakunthala , Asgharzadeh Shahab
Published: August 2019

Background: Anti-disialoganglioside (GD2) specific CAR T-cell therapy has shown limited efficacy in children with recurrent/refractory neuroblastoma (NBL). A recent pre-clinical study demonstrated significant neurotoxicity in mice treated with CAR T-cells bearing a high-affinity anti-GD2 short chain variable fragments (scFv; Richman S et al. 2017). To devise a safe and more efficacious CAR T-cells, we developed a gated CAR T-cell system utilizing the Syn-Notch technology.

Methods: GD2 and CD19 specific Syn-Notch gated systems were constructed by linking corresponding scFv to the Notch core followed by Gal4Vp64. Upon target recognition, Gal4VP64-dependent UAS element would initiate expression of the B7H3-BBZ CAR. Reporter activity of CD19 or GD2 Syn-Notch systems were monitored using flow cytometry after exposure to CD19+ transduced CHLA255 NBL cell line. In vitro cytotoxicity was assessed using DIMSCAN and xCELLigence impedance assays at varying Effector: Target ratios. NSG mice with established metastatic CHLA255(GD2+CD19+) NBL were treated with 1x10^7 transduced and control T cells and monitored for disease burden.

Results and discussion: Jurkat and human primary T cells were successfully transduced with the Syn-Notch system. GD2-Notch-Gal4VP64 and CD19-Notch-Gal4VP64 T-cells effectively activated downstream reporter in exposure to CHLA255(GD2+CD19+) within 6 hours of exposure but not in the presence of NBL cells lacking GD2 or CD19. The GD2 system exhibited greater leakiness than the CD19 counterpart. GD2- Syn-Notch-B7H3-BBZ or CD19- Syn-Notch-B7H3-BBZ CAR T-cells demonstrated statistically significant cytotoxicity against CHLA255(GD2+CD19+) NBL cell line. Preliminary in vivo data also show the anti-tumor effect of these gated CAR systems compared to mock-transduced T-cells.

Conclusions: We have successfully established a novel GD2-B7H3 gated system using the Syn-Notch technology, and demonstrate successful specificity to both antigen targets. This system increases the specificity of CAR T-cells against GD2 and B7H3 positive neuroblastoma tumors and likely minimize GD2-induced toxicity. These gated CARs show great promise for establishing immunotherapies that maximize tumor specificity while minimizing toxicity.

Citation Format: Babak Moghimi, Sakunthala Muthugounder, Shahab Asgharzadeh. Improving efficacy and accuracy of CAR-T cells for neuroblastoma [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 1437.
Abstract 1442: Chimeric antigen receptor T cell therapy targeting ICAM-1 in gastric cancer
Authors: Jung Minkyu , Zaman Marjan , Vedvyas Ygindra , Zhang Xianglan , McCloskey Jaclyn E. , Yang Yanping , Min Irene M. , Zamegar Raza , Choi Yoon Young , Cheong Jae-Ho , Noh Sung Hoon , Rha Sun Young , Chung Hyun Cheol , Jin Moonsoo M.
Published: August 2019

Introduction:

Despite advances in treatment, gastric cancer (GC) remains among the most fatal malignancies. Intercellular adhesion molecule-1 (ICAM-1) is overexpressed and associated with various cancers, including GC. We developed a third-generation chimeric antigen receptor (CAR) targeting ICAM-1 (ICAM-1-CAR) and investigated ICAM-1 as a immunotherapeutic target for CAR T therapy in GC.

Methods:

We investigated ICAM-1 expression in tissues of curatively resected GC patients (n=134) by immunohistochemical staining to determine its prognostic value. Effector to target assays were performed with ICAM-1-CAR T cells co-cultured with 8 GC cell lines with varying levels of ICAM-1 expression to investigate specific target cell death. We created a firefly luciferase-expressing human GC model in mice to measure tumor growth and killing by whole body bioluminescence imaging. To find the best treatment route and optimal dose of T cells in GC mice model, ICAM-1-CAR was injected via tail vein and intraperitoneal administration (IP) at two different doses, i.e., low dose (1 million CAR T) and high dose (10 million live T cells).

Results:

The ICAM-1 expression was higher in advanced stages (22.2% in stage II vs. 48.8% in stage III, p=0.002) and the patients with high ICAM-1 expression showed significantly poor survival (disease free survival, hazard ration [HR], 4.55, p<0.001; overall survival, HR, 3.89, p<0.001). The efficacy of ICAM-1-CAR T cells in vitro showed a strong correlation with the level of ICAM-1 expression in target cells, i.e., faster killing of GC with higher ICAM-1 expression. ICAM-1-CART facilitated regression of tumor in MKN-28 xenograft IP model. Additionally, the efficacy of ICAM-1-CAR was more prominent in mice treated with high dose of T cell and IP route. Compared to no treatment, ICAM-1CAR via IP led to tumor reduction that persisted for over 80 days and significantly improved survival without toxicity (p=0.049).

Conclusion:

ICAM-1 specific CAR T cells demonstrated significant therapeutic efficacy in vitro and in vivo against preclinical GC models. ICAM-1-CAR T cells may be developed into a promising treatment strategy for patients with ICAM-1 positive GC cancers.

Citation Format: Minkyu Jung, Marjan Zaman, Ygindra Vedvyas, Xianglan Zhang, Jaclyn E. McCloskey, Yanping Yang, Irene M. Min, Raza Zamegar, Yoon Young Choi, Jae-Ho Cheong, Sung Hoon Noh, Sun Young Rha, Hyun Cheol Chung, Moonsoo M. Jin. Chimeric antigen receptor T cell therapy targeting ICAM-1 in gastric 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 1442.
Abstract 1469: Optimizing the tolerability of intravenous oncolytic viral immunotherapy administration: investigation of a low-high-high dose schedule of enadenotucirev (EnAd) administration
Authors: McElwaine-Johnn Hilary , Illingworth Sam , Lear Rochelle , Brown Richard , Champion Brian
Published: August 2019

Background:EnAd is a tumor-selective oncolytic adenovirus with highly selective replication and cell killing in a broad range of carcinoma cell lines. Following intravenous dosing in clinical studies, uptake and replication of EnAd, associated with CD8+ cell infiltration, has been shown. It is currently in combination studies with nivolumab and paclitaxel. Intravenous delivery of EnAd stimulates an acute response characterised by an increase in cytokines in peripheral blood. As previously reported (ESMO IO 2017, abstr 203), the MTD of EnAd (3x1012 viral particles (vp) Days 1, 3 and 5, flat dosing schedule) has been determined primarily based on AEs occurring on first dose (C1D1), with the cytokine response, including interleukin-6 (IL-6), to subsequent doses of EnAd appearing attenuated when delivered within a few days of the first dose.

Methods: The effect of the first EnAd dose on subsequent acute particle-mediated cytokine (IL-6) responses in mice was investigated following administration of systemic doses equivalent to between 5.5 x 1012 vp and 5.5 x 1013 vp in humans.

Results: The magnitude of IL-6 response to a low first dose, which itself induces very low or undetectable levels of IL-6 cytokine, is sufficient to attenuate the response to subsequent high doses (10 times more vp). When administered as first dose these high doses induced a significant elevation in systemic IL-6.

Conclusions: Tolerability of EnAd is primarily determined by C1D1, correlating with the cytokine response across the patient cohort. The data from this in vivo pharmacology study support the concept of delivering a lower dose of enadenotucirev on Day 1 and administering higher doses thereafter, with the aim to deliver a higher viral load to tumour tissue and potentially optimise the overall risk:benefit of EnAd administration. This type of dosing regimen (low-high-high) has been successfully implemented in clinical studies of other systemically administered oncolytic viruses (Clin Cancer Res 2006;12:2555-2562) and is under clinical investigation as part of the ongoing combination study with nivolumab. (EudraCT:2013-001276-38).

Citation Format: Hilary McElwaine-Johnn, Sam Illingworth, Rochelle Lear, Richard Brown, Brian Champion. Optimizing the tolerability of intravenous oncolytic viral immunotherapy administration: investigation of a low-high-high dose schedule of enadenotucirev (EnAd) administration [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 1469.
Abstract 148: Analysis of the tumor microenvironment in seven cancer types by single-cell RNA-seq
Authors: Dean Michael , Li Guibo , Jin Jianhua , Hou Yong , Wu Kui , Zhu Shida , Zhou Hanlin , Lv Ruqian , Lin Feng , Liu Si , Dong Shichen , Wang Lei , Zhang Cuijuan , Zhao Yi
Published: August 2019

The tumor microenvironment (TME) is composed of tumor cells, as well as immune and stromal cells. Cancer cells can interact with the tumor microenvironment to suppress anticancer immunity and host inflammatory cells can modulate that immune response to the lesion. To better understand the TME across cancer types we used single-cell RNA-seq analysis of 224,330 cells from 77 patients with breast, gastric, nasopharyngeal carcinoma (NPC), diffuse large B-cell lymphoma (DLBCL), pancreatic, colorectal and lung cancer. The tumor cells and immune cells from individual patients were analyzed simultaneously at the single-cell level to identify distinct cell clusters and subtypes. Comparison of the profile of T cells, macrophages, dendritic cells (DC), B cells, mast cells, granulocytes and cancer-associated fibroblasts (CAFs) identifies unique profiles by tumor type. Breast tumors have high numbers of macrophages, colorectal tumors are elevated for CD4+-T cells, DLBCLs are low in CAFs, gastric cancers have high levels of mast cells, lung tumors high levels of CD4+-T cells and natural killer (NK) T cells, NPC have elevated levels of B cells and DCs and pancreatic cancers high levels of CAFs and granulocytes and low levels of DC and CD8+-T cells. However, within a tumor type, there is also considerable heterogeneity. Understanding how the TME evolves and changes with therapy or predicts response to treatment and survival may provide insight into tumorigenesis and cancer progression.

Note: This abstract was not presented at the meeting.

Citation Format: Michael Dean, Guibo Li, Jianhua Jin, Yong Hou, Kui Wu, Shida Zhu, Hanlin Zhou, Ruqian Lv, Feng Lin, Si Liu, Shichen Dong, Lei Wang, Cuijuan Zhang, Yi Zhao. Analysis of the tumor microenvironment in seven cancer types by single-cell RNA-seq [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 148.
Abstract 155: Cysteine-rich secretory protein 3 expression leads to invasive prostate cancer by modulating cell motility
Authors: Furic Luc , Volpert Marianna , Hu Jinghua , O'Connor Anne , Rebello Richard J. , Keerthikumar Shivakumar , Evans Jemma , Merriner Jo , Pedersen John , Risbridger Gail P. , McIntyre Peter , O'Bryan Moira K.
Published: August 2019

Cysteine-rich secretory protein 3 (CRISP3) is one of the most highly up-regulated proteins during the transition from a healthy human prostatic epithelium to prostate cancer. The role of CRISP3 within this process has not however, been defined. Here we show using a genetically engineered mouse model of prostate cancer, that CRISP3 production greatly facilitates disease progression from carcinoma in situ to invasive prostate cancer in vivo. This observation was further validated using both human and mouse prostate cancer cell lines, which showed that exposure to CRISP3 enhanced cell motility and invasion. Further,using mass spectrometry, we showed that this activity is induced, at least in part, via changes in cell-cell adhesion proteins, including LASP1 and TJP1 both in vivo and in vitro. Collectively, these data identify CRISP3 as being pro-tumorigenic in the prostate and validate it as a bona fide marker of aggressive prostate cancer and a potential target for therapeutic intervention.

Citation Format: Luc Furic, Marianna Volpert, Jinghua Hu, Anne O'Connor, Richard J. Rebello, Shivakumar Keerthikumar, Jemma Evans, Jo Merriner, John Pedersen, Gail P. Risbridger, Peter McIntyre, Moira K. O'Bryan. Cysteine-rich secretory protein 3 expression leads to invasive prostate cancer by modulating cell motility [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 155.
Abstract 1564: Polymorphisms in nucleotide excision repair genes were associated with the survival of post-operative oral squamous cell carcinoma patients treated with adjuvant concurrent chemoradiotherapy
Authors: Yeh Chih-Ching , Senghore Thomas , Chien Huei-Tzu , Wang Wen-Chang , Chen You-Xin , Young Chi-Kuan , Huang Shiang-Fu
Published: August 2019

Background: The nucleotide excision repair (NER) pathway plays a major role in the repair of DNA damaged by exogenous agents including chemo-and radiotherapeutic agents. Thus, we investigate the association between key potentially functional single nucleotide polymorphisms (SNPs) in the NER pathway and clinical outcomes in oral squamous cell carcinoma (OSCC) patients treated with concurrent chemoradiotherapy (CCRT).

Methods: Thirteen SNPs in five key NER genes were genotypes in 319 OSCC patients using iPLEX MassARRAY. Multivariate Cox proportional hazards models and Kaplan-Meier survival curves were used to estimate risk of death or recurrence.

Results: Carriers of the XPC rs2228000 TT genotype showed a borderline significant increased risk for poor overall survival under the recessive model (hazard ratio [HR]=1.81, 95% confidence interval [CI]=0.99–3.29). The CC genotypes of ERCC5 rs17655 (HR=1.54, 95% CI=1.03–2.29) and ERCC1 rs735482 (HR=1.65, 95% CI=1.06–2.58) were associated with an increased risk for worse disease-free survival under the recessive model. In addition, participants carrying both CC genotype of ERCC5 rs17655 and ERCC1 rs735482 exhibited an enhanced susceptibility for recurrence (HR=2.60, 95% CI=1.11–6.09). However, no statistically significant interaction was observed between them.

Conclusion: Our findings show that the ERCC5 rs17655 CC and ERCC1 rs735482 CC genotypes were associated with an increased risk for recurrence in OSCC patients treated with CCRT. Therefore, CCRT may not be beneficial and alternative treatments are required for them.

Citation Format: Chih-Ching Yeh, Thomas Senghore, Huei-Tzu Chien, Wen-Chang Wang, You-Xin Chen, Chi-Kuan Young, Shiang-Fu Huang. Polymorphisms in nucleotide excision repair genes were associated with the survival of post-operative oral squamous cell carcinoma patients treated with adjuvant concurrent chemoradiotherapy [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 1564.
Abstract 1567: Race, RNA-based recurrence scores, and survival of early hormone receptor-positive breast cancer: A population-based study
Authors: Han Yunan , Colditz Graham A. , Liu Ying
Published: August 2019

Introduction: Compared with white women, black American women are less likely to develop hormone receptor-positive breast cancer, but experience higher mortality of the disease. The 21-gene recurrence scores (RS) predict individualized benefit of chemotherapy for early hormone receptor-positive breast cancer. We hypothesize that the RS scores in early hormone receptor-positive breast cancer are higher in black women than in white women and this contributes to the worse outcomes among black women with hormone receptor-positive breast cancer.

Methods: Utilizing the Surveillance, Epidemiology, and End Results database, we identified 75,861 women (75.7% non-Hispanic white, 7.9% non-Hispanic black, 8.0% Asian/Pacific Islander, and 8.5% Hispanic) with stage I-III hormone receptor-positive breast cancer diagnosed between 2004 and 2014. RS scores were categorized into low, medium, and high groups using traditional cutoffs (18 and 30) and the cutoffs used in the TAILORx clinical trial (11 and 25). We performed multivariable logistic regression to evaluate odds ratios (ORs) of RS categories, and Cox proportional hazard regression to estimate hazard ratios (HRs) for breast cancer-specific mortality and overall mortality.

Results: The multivariable adjusted least square mean of RS was significantly higher in black (23.2) than other racial/ethnic groups (white 22.9, Asian/PI 22.5, Hispanic 22.4; p<0.0001). Compared with white women, black women were more frequently in the high RS category defined by a cutoff of 30 (case-only OR=1.24, 95% CI: 1.11-1.39), Asian/PI (OR=0.88, 95% CI: 0.79-0.98) and Hispanic (OR=0.80, 95% CI: 0.72-0.89) were less frequently in the high RS category defined by a cutoff of 25. Compared with white women, the HR of breast cancer-specific mortality was 1.37 (95% CI: 1.10-1.69) in black women, and 0.65 (95% CI: 0.48-0.90) in Asian/PI women, after adjustment for demographic factors, RS categories and other tumor characteristics, and treatment. This association did not significantly vary by RS categories. The multivariable-adjusted associations between race/ethnicity and overall mortality were similar to those for breast cancer-specific mortality.

Conclusions: Racial/ethnic differences in early hormone receptor-positive breast cancer outcomes could not be explained by RS scores. Potential differences in tumor biology that are not related to the genes in the 21-gene assay may contribute to the worse prognosis of black women with hormone receptor-positive breast cancer.

Citation Format: Yunan Han, Graham A. Colditz, Ying Liu. Race, RNA-based recurrence scores, and survival of early hormone receptor-positive breast cancer: A population-based 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 1567.
Abstract 1570: Genome-wide association study identifies variants at 3p26.1 linked to survival in ovarian cancer patients among Chinese women
Authors: Dai Hongji , Chu Xinlei , Wang Wei , Yang Huijun , Wang Qiong , Li Mulin Jun , Wei Qingyi , Chen Kexin
Published: August 2019

Survival rates for ovarian cancer varies considerably among patients, and some of this variation could be explained by germline genetic variation. In order to identify genetic markers associated with ovarian cancer-specific survival, we performed a genome-wide association study (GWAS) of ovarian cancer survival among Chinese women. In a meta-analysis of 2,130 ovarian cancer patients from the southeast and north part of China, we found a significant association between SNPs at the 3p26.1 and overall survival (lead SNP rs3804994; combined P=9.8×10-9). Patients with the minor allele are at increased risk for mortality (HR: 1.55; 95% CI: 1.33-1.80) relative to patients with the major allele. Statistical fine-mapping and functional prioritization recommend a potential casual SNP rs9311399 that locates in the active chromatin region of ovarian tissue. Using EMSA, luciferase assay and ATAC-seq, we demonstrated that risk allele of rs9311399 could alter the regulatory activity of DNA sequence. Our findings suggest that genetic variants at the 3p26.1 may be promising prognostic biomarkers for ovarian cancer patients.

Citation Format: Hongji Dai, Xinlei Chu, Wei Wang, Huijun Yang, Qiong Wang, Mulin Jun Li, Qingyi Wei, Kexin Chen. Genome-wide association study identifies variants at 3p26.1 linked to survival in ovarian cancer patients among Chinese 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 1570.
Abstract 1575: DNA methylation-derived neutrophil-to-lymphocyte ratio and survival among pediatric medulloblastoma patients
Authors: Arroyo Vidal M. , Lupo Philip J. , Scheurer Michael E. , Rednam Surya P. , Murray Jeffrey C. , Okcu M F. , Chintagumpala Murali M. , Brown Austin L.
Published: August 2019

Background/Objectives: Medulloblastoma is the most common malignant brain tumor diagnosed in children. Current risk-stratification methods of pediatric medulloblastoma do not fully explain the observed variability in clinical outcomes. Methylation-derived neutrophil-to-lymphocyte ratio (mdNLR) captures immune-specific information and has been identified as a potential prognostic biomarker of outcomes in various cancers, including adult brain tumors. Therefore, we evaluated the association between blood-derived mdNLR and overall survival in a cohort of patients with pediatric medulloblastoma.

Design/Methods: We identified pediatric patients diagnosed and treated for medulloblastoma at Texas Children’s Cancer Center between 1995 and 2015. Peripheral blood DNA methylation was measured using the Infinium Human Methylation 450K Beadchip. Methylation data underwent quality control, including beta-mixture quantile normalization and batch correction. Immune cell proportions (CD4+ T-Cells, CD8+ T-Cells, B-cells, natural killer cells, monocytes, and granulocytes) were estimated using cellular deconvolution methods and used to estimate mdNLR, which was then log-transformed to improve normality. Cox regression models were estimated to evaluate the association between mdNLR and overall survival.

Results: Of the 78 eligible patients include in this analysis, 83% (n=65) were alive at last follow-up (median follow-up=7.8 years). Deceased patients (n=13; median follow-up=2.6 years) had a higher mean mdNLR than patients who were alive at last contact (12.3 vs. 4.0, P = 0.046). Elevated log-transformed mdNLR was associated with an increased risk of death in both unadjusted models (HR=1.68, 95%CI: 1.11-2.55) and models accounting for age, sex, race, and clinical risk group (HR=1.97, 95%CI: 1.12-3.45).

Conclusion: We identified a significant association between peripheral blood mdNLR and survival in pediatric medulloblastoma. As a promising prognostic biomarker of outcomes, mdNLR captures immune-specific information and is a potential avenue of research in settings where cytologic determination of NLR may not be possible. Future work should investigate the relationship between elevated mdNLR and specific pediatric medulloblastoma molecular subtypes.

Citation Format: Vidal M. Arroyo, Philip J. Lupo, Michael E. Scheurer, Surya P. Rednam, Jeffrey C. Murray, M F. Okcu, Murali M. Chintagumpala, Austin L. Brown. DNA methylation-derived neutrophil-to-lymphocyte ratio and survival among pediatric medulloblastoma 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 1575.
Abstract 158: Mutantp53, MAPK and STAT3 promote metastasis to the brain: Demonstrated by protein, gene, and functional analysis
Authors: Wainwright John V. , Cooper Jared B. , Chandy Anisha , Gandhi Chirag D. , Schmidt Meic H. , Jhanwar-Uniyal Meena
Published: August 2019

Metastatic tumors to the brain arise from a specific subset of neoplastic cells originating from primary tumors such as lung, breast, colon, etc which disseminate specifically to the cerebral gray/white matter junction. Genetic and epigenetic mechanisms of this complex metastatic transformation are yet to be defined. Inactivation of the tumor suppressor gene p53 is one of the most frequent genetic alterations seen in human cancers. Further, extracellular regulated kinase (ERK), a component of mitogen activated protein kinase (MAPK) and Signal transducer and activator of transcription 3 (STAT3) plays a crucial role in differentiation, proliferation and migration, and is aberrant in many tumors. We hypothesize that aberrant p53, ERK1/2 and STAT3 contribute to metastatic brain tumors. Results demonstrated that: 1. Immunohistochemical analysis (IHC) of mutant p53 (mt-p53), phosphorylated-ERK1/2 Thr202/Tyr204 (pERK 1/2) and phosphorylated-STAT3Tyr705 revealed that 66% of metastatic brain tumors expressed mt-p53, 86% of tumors expressed pERK 1/2, and 75% nuclear expression of pSTAT3Tyr705; 2. Gene expression profiling showed that 17, 44 and 21 differentially expressed genes in metastatic tumors were p53, MAPK and STAT3 associated, respectively with alterations in genes associated with cell cycle regulation, neurogenesis, and differentiation and reprogramming and metastases; and 3. Metastatic breast cancer cells grown in astrocytic media displayed increased cell proliferation and enhanced S-phase cell cycle entry. Migration of primary tumor cells was significantly enhanced in astrocytic media as well. Inhibitors of MAPK (U0126) and STAT3 (STAT3 inhibitor SVI-201) suppressed cell proliferation and migration. Metastatic tumor cells and astrocytes were shown to grow amicably together, forming cell-to-cell interactions. These findings suggest that mt-p53 and aberrant MAPK and STAT3 play a crucial role in brain metastasis, and that the cerebral milieu provides a suitable microenvironment for metastatic cells to grow and disseminate and inhibition of aforementioned pathway can provide suitable therapeutic strategies.

Citation Format: John V. Wainwright, Jared B. Cooper, Anisha Chandy, Chirag D. Gandhi, Meic H. Schmidt, Meena Jhanwar-Uniyal. Mutantp53, MAPK and STAT3 promote metastasis to the brain: Demonstrated by protein, gene, and functional analysis [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 158.
Abstract 1593: Genetic interactions in lung cancer using machine-learning approaches in genome-wide association studies
Authors: Byun Jinyoung , Han Younghun , Amos Christopher I.
Published: August 2019

Genome-wide association studies (GWAS) consisting of hundreds of thousands or millions of single nucleotide polymorphisms (SNPs) have presented the complex inheritance patterns of disease/trait. Although GWAS have identified thousands of SNPS associated with diseases, few studies have explored interactions among these SNPs that may influence disease risks. Machine learning applications can identify SNPs that influence disease risk through interactions and define how these SNPs jointly influence disease risks. Tree-based machine-learning applications; classification and regression trees (CART) and random forest (RF) methods, have become increasingly popular and convenient tools for understanding interactions influencing disease development. Here we apply these methods to understand the genetic architecture of lung cancer. To elucidate the SNP-SNP interactions that influence lung cancer risk, we applied supervised tree-based approaches using 18,444 cases and 14,027 controls from lung cancer OncoArray GWAS data that is the largest lung cancer GWAS so far. To reduce the space of SNPs to consider in modeling, we first selected the SNPs very significantly (p<0.00001) associated with lung cancer risk. Random Forests, which consists of systematically fitting classification trees, was run 1,000 times to identify the most influential SNPs that jointly influence lung cancer risk. Subsequently we applied a classification tree approach to summarize interactions that predict risk. The final parsimonious tree included effects from genetic variants in rs55781567(CHRNA5): rs452384(CLPTM1L): rs9258608(LOC105375010): rs6154144(DNAJC5): rs116506680(HLA-G): rs9271365(near HLADQA1): rs421629(CLPTM1L). The nodes of this tree had Odds ratios for lung cancer ranging from 0.87 to 1.63. Machine-learning approaches in genomics can provide an important genetic mechanism in lung cancer development.

Citation Format: Jinyoung Byun, Younghun Han, Christopher I. Amos. Genetic interactions in lung cancer using machine-learning approaches in genome-wide association studies [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 1593.
Abstract 1594: Transcriptome-wide association study analysis of nasopharyngeal carcinoma
Authors: Su Wen-Hui , Chan Fu-Chun
Published: August 2019

Arising from the epithelial cells in nasopharynx, nasopharyngeal carcinoma (NPC) is highly prevalent in Southeastern China, including Hong Kong and Taiwan. Genetic and environmental factors play important role in NPC susceptibility. Genome-wide association studies (GWASs) have identified single-nucleotide polymorphisms (SNPs) located in major histocompatibility complex (MHC) region with strong evidence for association with NPC. Recently, combination of GWAS and expression quantitative trait loci (eQTL) datasets, transcriptome-wide association study (TWAS) was used to identify novel associations in transcriptome level. In this study, we conducted TWAS in NPC using two in house GWAS datasets and GTEx eQTL datasets from public domain. Most of the significant results were located in chromosome six MHC region. Our analysis identified significant TWAS genes that explain the most of the GWAS signals : HLA-A, GABBR1 and HLA-F genes with P values of 5.08E-13, 5.78E-09 and 2.30E-08, respectively. One of the novel genes, ZNRD1 (P = 8.22 E-08), was known to be involved in the tumorigenesis of esophageal cancer and liver cancer. Further analysis should be needed to understand the mechanisms between ZNRD1 association in NPC.

Citation Format: Wen-Hui Su, Fu-Chun Chan. Transcriptome-wide association study analysis of nasopharyngeal carcinoma [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 1594.