bluuebunny/crossref_metadata_2025 huggingface dataset:
Abstract 6379: Dipeptidyl peptidase-4 inhibitor impairs the outcomes of patients with type 2 diabetes mellitus after curative resection for colorectal cancer
Authors: Saito Akira , Ohzawa Hideyuki , Kaneko Yuki , Tamura Kohei , Futoh Yurie , Takahashi Kazuya , Kimura Yuki , Tojo Mineyuki , Kawashima Rie , Miyato Hideyo , Sata Naohiro , Kitayama Joji
Published: June 2022

Background: Type2 diabetes mellitus(T2DM) is a risk factor for cancer. Recent studies have shown that DPP-4 inhibitor(DPP-4i) can either promote or suppress cancer progression. However, the detailed mechanisms remain unknown. Here, in this study, we investigated the effect of DPP-4i on tumor microenvironment and its effect on the prognosis of cancer patients.

Method: We retrospectively examined the outcome of colorectal cancer (CRC) patients with T2DM who received curative surgery in Jichi Medical University Hospital and asked the impact of DPP-4i intake on their outcome. In addition, we performed immunohistochemistry to examine the phenotypes of TIL, TAM and epithelial-mesenchymal transition (EMT) of cancer cells in surgically removed CRC tissues in 40 CRC patients who had taken DPP-4i and propensity score-matched 40 patients who had not.

Results: A total of 1696 patients underwent curative colectomy from April 2010 to March 2020. Among them, 260 patients had T2DM at the time of surgery, and 135 patients had been treated with drugs including DPP-4i. The postoperative disease-free survival rate (DFS) was significantly worse in the DPP-4i intake group compared with non-intake group (5 year DFS 18.5% vs 5.4%, HR=2.0, p<0.05). The number of Zeb1-positive tumor cells significantly increased in DPP-4i intake group. (Median(M)=29.0 (0-189)/mm2 vs 9.0 (0-71)/mm2, p<0.01). The number of CD3 (+) TIL in DPP-4i intake group was Median(M)=277.4(min 121.6-max 523.2)/mm2 which was significantly lower than those in non-intake group (M= 319.6, min 493.0-max 823.6/mm2). CD8(+) TIL in DPP-4i intake was reduced more significantly than in non-intake group (M=187.6, min 67.8-max 347.4/mm2 vs M=336.8, min 200.2-max 588.4/mm2, p<0.05) with less CD8/CD3 ratio (61.2% vs 67.1%, p<0.05). On the other hand, the density of CD68(+)CD163(+) TAM was significantly higher(M=202.6, min 122.8-max 257.0/mm2 vs M=126.6, min 94.8-max 247.6/mm2, p<0.05) in DPP-4i intake group.

Conclusion: DPP-4i increases the number of M2-type TAM while decreases the number of effector TIL and promote EMT of cancer cells in tumor microenvironment, which might lead to the poor outcome of the patients with CRC.

Citation Format: Akira Saito, Hideyuki Ohzawa, Yuki Kaneko, Kohei Tamura, Yurie Futoh, Kazuya Takahashi, Yuki Kimura, Mineyuki Tojo, Rie Kawashima, Hideyo Miyato, Naohiro Sata, Joji Kitayama. Dipeptidyl peptidase-4 inhibitor impairs the outcomes of patients with type 2 diabetes mellitus after curative resection for colorectal cancer [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2022; 2022 Apr 8-13. Philadelphia (PA): AACR; Cancer Res 2022;82(12_Suppl):Abstract nr 6379.
Abstract 6394: The SB Image Processing Toolkit: Machine learning for cancer research on the CGC
Authors: Koc Soner , Babic Jovana , Nikolic Nevena , Stankovic Ana , Ventre Daniel , Ray Manisha , Worman Zelia F. , Subramanian Sai Lakshmi , Dean Dennis A. , DiGiovanna Jack , Dusenbery Brandi Davis
Published: June 2022

Applying machine learning algorithms to analysis of multi-omics datasets has provided a wealth of analysis and interpretations of data not easily achievable by conventional methods. The Cancer Genomics Cloud (CGC), powered by Seven Bridges, offers many helpful features to simplify and streamline the performance of machine learning (ML) tasks on the platform. The CGC platform contains features and tools for all of the steps of a ML project, from data exploration, to model generation, to production, so that all key steps occur on the platform. The Data Cruncher interactive analysis tool enables users to perform data exploration, modeling, training, and visualization using familiar frameworks like Jupyterlab and RStudio. We applied this ML methodology to imaging tools, to create The SB Image Processing Toolkit. Here we demonstrate the use of this toolkit with specific cancer datasets, including training and analysis, with all steps taking place on the cloud.

The SB Image Processing Toolkit is a collection of various deep-learning, preprocessing, and utility tools and workflows created with the purpose of performing image class prediction on any type of image data. The toolkit supports a variety of images used in cancer research, such as histopathology and radiology images, and common formats like JPG or PNG. There are nine distinct tools within the toolkit, including methods for quality control, normalization, preprocessing for histopathology or radiology images, classification, and deep learning. Together, these tools create an easy-to-use infrastructure to enable various stages of ML and Image Processing. Most importantly, it was designed to enable users to create and use ML image classifiers on the CGC without any coding experience. All workflows can be searched for and implemented via the Public Apps Gallery on the CGC. Users can import image data, run analysis, and compare results all in one environment.

By harnessing the scale and flexibility of cloud computing, the SB Image Processing Toolkit brings massive speed and price improvements compared to manual writing and running model training scripts. In conjunction with the access to data and ease of use, the CGC makes complex machine learning methods accessible to any researcher.

Citation Format: Soner Koc, Jovana Babic, Nevena Nikolic, Ana Stankovic, Daniel Ventre, Manisha Ray, Zelia F. Worman, Sai Lakshmi Subramanian, Dennis A. Dean, Jack DiGiovanna, Brandi Davis Dusenbery. The SB Image Processing Toolkit: Machine learning for cancer research on the CGC [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2022; 2022 Apr 8-13. Philadelphia (PA): AACR; Cancer Res 2022;82(12_Suppl):Abstract nr 6394.
Abstract 642: Spatial analysis of tumor infiltrating lymphocytes in histological images using deep learning predicts progression-free survival in colorectal cancer
Authors: Xu Hongming , Cha Yoon Jin , Clemenceau Jean , Choi Jinhwan , Lee Sung Hak , Kang Jeonghyun , Hwang Tae Hyun
Published: June 2022

Histological distribution and density of tumor infiltrating lymphocytes (TILs) are known to be informative to cancer patients’ outcome. However, due to the heterogeneity and huge size of digitized pathology slides, it has been a challenging problem to quantify TILs-related features in whole slide images (WSI). This study aimed to explore the impact of spatial distribution of TILs quantified by deep learning (DL) approaches based on digitalized WSI stained with hematoxylin and eosin in patients with colorectal cancer (CRC). The cancer and TILs regions are first separately identified by deep learning models in the WSI. The TILs densities at tumor invasive margins are then quantified by image analysis algorithms and used for prognosis of progression-free survival (PFS) in CRC patients. The prognostic impact of spatial distributions of TILs in CRC patients was explored in the Yonsei cohort (n=180) and validated in the TCGA cohort (n=268). Two experienced pathologists manually measured TILs at the most invasive margin as 0-3 by the Klintrup-Mäkinen (KM) grading method and compared to DL approaches. On multivariate analysis, TILs densities within 200µm of the invasive margin (f_im200) was remained as the most significant prognostic factor for PFS (HR 0.004 [95% CI, 0.0001-0.1502], p=.002) in the Yonsei cohort. On multivariate analysis using the TCGA dataset, f_im200 retained prognostic significance for PFS (HR 0.031, [95% CI 0.001-0.645], p=.024). Inter-observer agreement of manual KM grading was insignificant in both cohorts. The survival analysis by pathologists’ manual KM grading showed statistically significant different PFS from the TCGA cohort, but not the Yonsei cohort. In summary, automated quantification of TILs at the invasive margin showed a prognostic utility to predict PFS, and could provide reproducible TILs density measurement in CRC patients.

Citation Format: Hongming Xu, Yoon Jin Cha, Jean Clemenceau, Jinhwan Choi, Sung Hak Lee, Jeonghyun Kang, Tae Hyun Hwang. Spatial analysis of tumor infiltrating lymphocytes in histological images using deep learning predicts progression-free survival in colorectal cancer [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2022; 2022 Apr 8-13. Philadelphia (PA): AACR; Cancer Res 2022;82(12_Suppl):Abstract nr 642.
Abstract 655: Macrophage mediated resistance to TKI therapy in ALK fusion positive non-small cell lung cancer
Authors: Mandell Melanie , Priest Katherine , Le Anh , Heasley Lynn , Nemenoff Raphael , Schenk Erin L.
Published: June 2022

Patients with lung cancer bearing driver oncogenes such as epidermal growth factor receptor (EGFR) mutations or anaplastic lymphoma kinase (ALK) fusions, often experience long term disease control on targeted therapy. Despite excellent initial responses, ultimately these tumors develop resistance to targeted therapy. While several cancer cell intrinsic mechanisms of targeted therapy resistance are well known, they do not account for all cases of resistance. Based on our previous observations, we hypothesized components of the tumor microenvironment, specifically monocytes and macrophages, also contribute to the development of targeted therapy resistance. We generated bone marrow derived macrophages from C57BL/6 mice and polarized them to an M0, M1, or M2 phenotype over the course of 6 days. At day 7, cells were plated in fresh media and after 2 days, the conditioned media (CM) was collected. Macrophage polarization was verified via qPCR. Murine cell lines, EA-1, EA-2, and EA-3 are unique, murine lung adenocarcinomas bearing an oncogenic ALK fusion derived from C57BL/6 mice. These murine cell lines were then cultured in the macrophage CM prior to exposure to ALK fusion specific tyrosine kinase inhibitors (TKI), alectinib or crizotinib. We observed that culture in M2-CM resulted in at least a 10-fold increase in alectinib resistance across all 3 murine cell lines when compared to the same cell lines cultured in M1 or M0-CM. We next tested for the activity of known, clinically relevant bypass signaling pathways as mediator of M2-CM protection. Aftatinib, an inhibitor of EGFR and human epidermal growth factor receptor 2 (HER2) family of proteins, did not alter M2-CM protection in the presence of alectinib. M2-CM did not impart TKI resistance to the murine cell lines in the presence of crizotinib, a TKI which inhibits ALK fusion activity and MET receptor tyrosine kinase activity, suggesting bypass signaling through MET drives the M2-CM resistance. Ongoing animal model studies demonstrate a clear role for innate immune cells in decreasing tumor responsiveness to TKI. In summary, our work demonstrates TKI therapy may be modulated by the presence of and interaction with specific immune cells within the tumor microenvironment. These data support exploring macrophage targeting therapies to improve TKI responsiveness.

Citation Format: Melanie Mandell, Katherine Priest, Anh Le, Lynn Heasley, Raphael Nemenoff, Erin L. Schenk. Macrophage mediated resistance to TKI therapy in ALK fusion positive non-small cell lung cancer [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2022; 2022 Apr 8-13. Philadelphia (PA): AACR; Cancer Res 2022;82(12_Suppl):Abstract nr 655.
Abstract 780: Transcriptional profile of human pancreatic acinar ductal metaplasia
Authors: Perkins Corey Melissa , Jiang Jinmai , Hakimjavadi Hesamedin , Bray Julie K. , Gosling Alyssa , da Silva Lais , Bulut Gamze , Ali Jamel , Setiawan Wendy , Campbell-Thompson Martha , Chamala Srikar , Schmittgen Thomas D.
Published: June 2022

Aberrant acinar to ductal metaplasia (ADM), one of the earliest events involved in exocrine pancreatic cancer development, is typically studied using pancreata from transgenic mouse models. We used primary, human pancreatic acinar cells to evaluate the transcriptional profile during the course of ADM. Following six days of culture on Matrigel, acinar cells underwent morphological and molecular changes reminiscent of ADM. RNA was sequenced from 14 donor’s paired pancreata (day 0 and 6 of culture). Unsupervised hierarchical clustering demonstrated complete separation of the gene expression profile between culture day 0 (acinar phenotype) and day 6 (ductal phenotype). By and large, acinar-specific genes were downregulated in the samples from day 6 ADM while ductal-specific genes were upregulated. Using a gene set enrichment approach, we identified regulons that are involved in regulating ADM including downregulated, acinar-associated transcription factors (including PTF1A, RBPJL, and XBP1) and upregulated, ductal- and progenitor-associated transcription factors (SOX11, SOX4, and YAP1). The expression of pancreatic cancer associated genes significantly correlated with the gene expression/regulon activity observed in normal pancreas undergoing ADM. We reported a detail analysis of the transcriptional profile during human ADM. Our findings confirm that many ADM-related transcription factors and signaling pathways discovered in transgenic mouse models are applicable to human ADM and highlights the relevancy of in vitro models of pancreas plasticity using human tissue.

Citation Format: Corey Melissa Perkins, Jinmai Jiang, Hesamedin Hakimjavadi, Julie K. Bray, Alyssa Gosling, Lais da Silva, Gamze Bulut, Jamel Ali, Wendy Setiawan, Martha Campbell-Thompson, Srikar Chamala, Thomas D. Schmittgen. Transcriptional profile of human pancreatic acinar ductal metaplasia [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2022; 2022 Apr 8-13. Philadelphia (PA): AACR; Cancer Res 2022;82(12_Suppl):Abstract nr 780.
Abstract 803: Identifying a RAD18/UBC13-dependent mechanism of replication fork recovery to modulate chemoresponse in BRCA1-deficient cancers
Authors: Cybulla Emily , Jackson Jessica , Tirman Stephanie , Quinet Annabel , Lemacon Delphine , Vindigni Alessandro
Published: June 2022

Mutations in the breast cancer susceptibility genes BRCA1 and BRCA2 are associated with an increased lifetime risk of breast and ovarian cancers. While the BRCA proteins play a well-established role in double-stranded DNA break repair, recent studies have revealed an emerging role of BRCA1/2 in replication stress response. While replication forks are extensively degraded by nucleases in BRCA-deficient cancer cells, activation of specialized fork recovery mechanisms enables resumption of DNA synthesis and promotes cell survival. My project aims to determine this fork recovery mechanism in BRCA1-deficient cells and to identify potential recovery factors that can be targeted to improve chemotherapeutic response in BRCA1-mutated breast and ovarian cancers. To monitor perturbations in replication fork dynamics on a genome-wide scale, we utilize a DNA fiber assay technique measuring rates of fork recovery and replication fork degradation. In parallel, electron microscopy analysis allows direct visualization of replication fork intermediates. Cell survival assays are employed to test how loss of fork recovery factors impacts cell proliferation and chemotherapeutic response in BRCA1-deficient cells. Our results reveal that RAD18 and UBC13, which catalyze ubiquitination of Proliferating Cellular Nuclear Antigen (PCNA), promote fork recovery in BRCA1-deficient, but not BRCA2-deficient, cancer cells. Previous work has also shown that PCNA polyubiquitination by UBC13 is important for reversed fork formation in BRCA-proficient cells. However, our findings show that extensive degradation of reversed fork substrates still occurs in BRCA1-deficient cells lacking RAD18 or UBC13, indicating that PCNA polyubiquitination is not essential for fork reversal in this genetic background. In addition, loss of RAD18 in BRCA1-deficient cells significantly slows cell proliferation, and UBC13 inhibition further sensitizes cells lacking BRCA1 to the replication stress inducer Hydroxyurea (HU). Based on our findings, we hypothesize that RAD18, UBC13, and PCNA ubiquitination may represent novel targets to improve chemoresponse in BRCA1-deficient cancers that rely on fork recovery mechanisms for survival.

Citation Format: Emily Cybulla, Jessica Jackson, Stephanie Tirman, Annabel Quinet, Delphine Lemacon, Alessandro Vindigni. Identifying a RAD18/UBC13-dependent mechanism of replication fork recovery to modulate chemoresponse in BRCA1-deficient cancers [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2022; 2022 Apr 8-13. Philadelphia (PA): AACR; Cancer Res 2022;82(12_Suppl):Abstract nr 803.
Abstract 817: microRNA-26a-5p is a prognostic factor that regulates cancer stemness
Authors: Kang Daeun , Park Chang Ryul , Lee Minhyeok , Lee Su Yel , Park Se Jin , Hwang Wan Jin , Ku Gwan Woo , Yu Seong Lan , Jeong In Beom , Kwon Sun Jung , Kang Jaeku , Lee Eung Bae , Son Ji Woong
Published: June 2022

Background: Cancer stem cells (CSCs) identified in lung cancer exhibit resistance to chemotherapy, radiotherapy, and targeted therapy. Therefore, a technology to control of CSCs is needed to overcome such resistance to cancer therapy. Various evidences about the association between epithelial-mesenchymal transition related transcriptomic alteration and acquisition of CSC phenotype have been proposed recently. In our previous research, down-regulated miR-26a-5p is closely related to mesenchymal-like lung cancer cell lines. These findings suggest that miR-26a-5p might be involved in lung cancer stemness.

Methods: RNA polymerase III subunit G (POLR3G) was selected as a candidate target of miR-26a-5p related to cancer stemness. its quantitative relationship was investigated by polymerase chain reaction, western blot after transfection of miR-26a-5p. luciferase assay were done for investigating the direct regulation of miR-26a-5p on POLR3G expression. After transfection of miR-26a-5p, colony formation assay and sphere formation assay were performed to evaluate the effect on cancer stemness. By treating cancer cell by miR-26a-5p and paclitaxel, cell viability was checked by 3-(4,5-dimethylthiazol-2-yl)-5-(3-carboxymethoxyphenyl)-2-(4-sulfophenyl)-2H-tetrazolium (MTS) assay and Muse cell analyzer. Expression level of each gene and its impact on survival were revealed by the cancer genome atlas pancancer database.

Results: miR-26a-5p regulated the expression of POLR3G directly. Overexpression of miR-26a-5p induced down regulation of POLR3G and a marked reduction of colony formation and sphere formation. Co-treatment of miR-26a-5p with paclitaxel decreased cell growth, suggesting that miR-26a-5p might play a role as a chemotherapy sensitizer. In the cancer genome atlas data, down-regulated miR-26a-5p and up-regulated POLR3G were shown compared to adjacent normal tissue. High miR-26a-5p and low POLR3G expression were also related to higher survival rate of patients with lung adenocarcinoma.

Conclusions: Overexpression of miR-26a-5p can suppress lung cancer stemness and make cancer cell become sensitive to chemotherapy. This finding provides a novel insight into a potential lung cancer treatment by regulating stemness.

Citation Format: Daeun Kang, Chang Ryul Park, Minhyeok Lee, Su Yel Lee, Se Jin Park, Wan Jin Hwang, Gwan Woo Ku, Seong Lan Yu, In Beom Jeong, Sun Jung Kwon, Jaeku Kang, Eung Bae Lee, Ji Woong Son. microRNA-26a-5p is a prognostic factor that regulates cancer stemness [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2022; 2022 Apr 8-13. Philadelphia (PA): AACR; Cancer Res 2022;82(12_Suppl):Abstract nr 817.
Abstract 830: ZFP14, a p53 target, suppresses tumor formation by regulating HOXA gene cluster expression
Authors: Mohibi Shakur , Zhang Jin , Chen Mingyi , Chen Xinbin
Published: June 2022

KRAB containing zinc finger proteins form the largest class of mammalian transcription factors. However, the physiological or pathological functions of the majority of them are unknown. Here, we established ZFP14 as a p53 target gene that is induced upon genotoxic stress in a p53-dependent manner. To determine the function of ZFP14 in mouse and human cancer cell lines, we generated ZFP14-KO cells that showed increased senescence as well as reduced cell growth and migration. Moreover, we showed that ZFP14 negatively regulates p53, which was the cause of reduced oncogenic properties in ZFP14-KO cancer cells. Mechanistically, we found that ZFP14 modulates p53 protein stability by associating with and possibly enhancing MDM2/p53 complex integrity through its zinc finger domains. Interestingly, we found that lower expression of ZFP14 correlates with poor patient survival in multiple human cancers. Notably, in a similar fashion, mice deficient in ZFP14 were susceptible to spontaneous tumor formation, developed other abnormalities such as systemic inflammation and liver steatosis, which led to their shorter lifespan. Finally, we showed that ZFP14 regulates the expression of the HOXA gene cluster in mouse and humans, which we propose as a possible mechanism by which ZFP14 regulates tumorigenesis. Our findings suggest that ZFP14 is an important regulator of p53 and HOXA genes during inflammation and tumorigenesis.

Citation Format: Shakur Mohibi, Jin Zhang, Mingyi Chen, Xinbin Chen. ZFP14, a p53 target, suppresses tumor formation by regulating HOXA gene cluster expression [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2022; 2022 Apr 8-13. Philadelphia (PA): AACR; Cancer Res 2022;82(12_Suppl):Abstract nr 830.
Abstract 834: Serine protease hepsin regulates tumor growth via TGFbeta-EGFR signaling axis
Authors: Tervonen Topi A. , Belitškin Denis , Munne Pauliina , Pant Shishir M. , Suleymanova Ilida , Belitškina Kati , Pouwels Jeroen , Klefström Juha
Published: June 2022

Hepsin (encoded by HPN gene) is a type II transmembrane serine protease, which is commonly overexpressed in carcinomas of prostate and breast. Hepsin protein is known to be stabilized by Ras-MAPK pathway signaling, and downstream, this protease regulates the degradation of extracellular matrix (ECM) components and activates growth factor pathways, including hepatocyte growth factor (HGF) and transforming growth factor beta (TGF beta) pathway. However, the impact of the hepsin-dependent signaling on cell proliferation and tumor growth is not well-understood. Therefore, we sought to clarify the role of hepsin during these events using engineered breast cancer cell lines, Hpn CRISPR knockout mouse model crossed with Wap-Myc breast cancer mouse model, and patient-derived explant cultures (PDEC)s from human breast tumors. We isolated Wap-Myc; Hpn-/- mammary tumor cells and made orthotopic transplantation into syngeneic wild-type recipient mice. The resulting Wap-Myc tumors that lack hepsin had reduced size both in primary and metastatic site compared to tumors derived from Wap-Myc; Hpn+/+ tumor cells. This decrease in growth was accompanied by downregulation in TGF beta and EGFR signaling as well as substantial reduction in total EGFR protein level. We further demonstrated that in 3D culture conditions overexpression of hepsin induced cell proliferation, which was TGF beta 1 and EGFR signaling-dependent, while PDECs treated with hepsin inhibitory antibodies and small molecules had decreased EGFR and TGF beta signaling activity and reduction in proliferation marker expression. Taken together, this study demonstrates a role for hepsin as a regulator of cell proliferation and tumor growth through TGF beta and EGFR pathways and may provide an interesting druggable upstream target to inhibit TGF beta and EGFR pathways in breast cancer.

Citation Format: Topi A. Tervonen, Denis Belitškin, Pauliina Munne, Shishir M. Pant, Ilida Suleymanova, Kati Belitškina, Jeroen Pouwels, Juha Klefström. Serine protease hepsin regulates tumor growth via TGFbeta-EGFR signaling axis [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2022; 2022 Apr 8-13. Philadelphia (PA): AACR; Cancer Res 2022;82(12_Suppl):Abstract nr 834.
Abstract 849: Characterization and prevention of radiation-induced malignancies in Li-Fraumeni Syndrome
Authors: Psarianos Pamela , Fischer Nicholas , Giovino Camilla , Ong Noel , Malkin David
Published: June 2022

PURPOSE: Li-Fraumeni Syndrome (LFS) is a genetic disorder associated with a significant risk of early-onset cancer. This condition is largely driven by germline mutations in the TP53 tumor suppressor, which has a broad spectrum of functions including the transcriptional regulation of radiation response. Termed the guardian of the genome, TP53 serves as a critical checkpoint which coordinates DNA repair, cell cycle progression and apoptosis following DNA damage, ultimately controlling the fate of the cell.

Aberrant or deficient TP53 function, such as occurs in LFS, contributes to radiation vulnerability. As a result, there is particular caution in the use of radiotherapy to treat primary LFS tumors, in order to prevent the formation of secondary, radiation-induced malignancies. To this end, therapeutic options for LFS are often limited to surgery and chemotherapy, posing substantial constraints for the treatment of patients who may otherwise benefit from primary tumor radiation.

Metformin, a commonly prescribed anti-diabetic drug, is associated with lower cancer incidence in populations worldwide. Recent studies have shed light on the potential utility of metformin as a pharmacopreventive agent for primary tumors in LFS; hence, we hypothesize that the utility of metformin can be extended to the prevention of secondary malignancies following radiotherapy in LFS.

METHODS/RESULTS: To explore the chemopreventive potential of metformin in LFS, we completed tumor challenges using a p53R172H/+ LFS mouse model. Mice treated prophylactically with metformin showed slower xenograft tumor growth compared to vehicle-treated mice. Ongoing work in our lab aims to characterize the effects of metformin on tumor growth following the administration of localized ionizing radiation (IR) to LFS mice. Briefly, these mice are treated with either vehicle or metformin, and IR is administered to the left hindlimbs to induce tumor formation. The timing and rates of tumor growth are monitored using magnetic resonance imaging. At endpoint, excised tumors are subjected to global profiling via RNA sequencing, proteomic analysis, and single-cell sequencing of discrete cell populations to characterize the effects of metformin on the development of radiation-induced malignancies. Results of these studies will be presented.

SIGNIFICANCE: Overall, this work will advance our understanding of the chemopreventive effects of metformin, with the potential to broaden the treatment options available to LFS patients.

Citation Format: Pamela Psarianos, Nicholas Fischer, Camilla Giovino, Noel Ong, David Malkin. Characterization and prevention of radiation-induced malignancies in Li-Fraumeni Syndrome [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2022; 2022 Apr 8-13. Philadelphia (PA): AACR; Cancer Res 2022;82(12_Suppl):Abstract nr 849.
Abstract 864: Ophiopogonin D increase apoptosis by activating p53 via ribosomal protein L5 and L11 and inhibiting the expression of c-Myc via CNOT2
Authors: Ko Hyun Min , Jee Wona , Park Do-il , Park Somi , Park Ye-Rin , Jang Hyeung-Jin , Jung Ji Hoon
Published: June 2022

Extracted from the root tuber of Ophiopogon japonicas, ophiopogon is well known to have an anti-cancer effect. However, the underlying mechanisms are still largely unknown. Here, we report that Ophiopogon D (OP-D) can inhibit colon cancer cell proliferation and induce apoptosis by inhibiting c-Myc expression through activation of p53 and CNOT2 regulation. Our results showed that OP-D induced p53 expression via ribosomal protein L5 or L11 and inhibited c-Myc expression through CNOT2 in a dose-dependent manner. Additionally, OP-D regulated cyclin D1 and CDK4 which are well known as cell cycle regulatory proteins. Consistently, OP-D inhibited the phosphorylation of AKT expression in a dose-dependent manner. Furthermore, OP-D shortened c-Myc’s half-life in a time-dependent manner. Furthermore, CNOT2 knockdown enhanced the inhibitory effect of OP-D on c-Myc in colon cancer cells. Interestingly, OP-D has increased the apoptotic effect of colon cancer cells when combined with doxorubicin or 5-FU, a treatment already used clinically. Altogether, our results suggested that OP-D regulates colon cancer cell survival and induces apoptosis by inhibiting c-Myc expression via activation of p53 and CNOT2 regulation.

Citation Format: Hyun Min Ko, Wona Jee, Do-il Park, Somi Park, Ye-Rin Park, Hyeung-Jin Jang, Ji Hoon Jung. Ophiopogonin D increase apoptosis by activating p53 via ribosomal protein L5 and L11 and inhibiting the expression of c-Myc via CNOT2 [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2022; 2022 Apr 8-13. Philadelphia (PA): AACR; Cancer Res 2022;82(12_Suppl):Abstract nr 864.
Abstract 899: Effects of innate immune deaminase deregulation on initiation and progression of myeloproliferative neoplasms
Authors: Isquith Jane Marie , Mark Adam , Pham Jessica , Donohoe Mary , Ladel Luisa , Jamieson Catriona
Published: June 2022

Inflammatory cytokine responsive APOBEC3 cytidine deaminases have been studied extensively with regard to innate immunity and more recently during cancer evolution. However, the mechanisms by which the APOBEC3 enzymes promote cancer initiation and progression in the malignant microenvironment remains to be investigated, especially in hematopoietic malignancies. Through whole genome and whole transcriptome sequencing analyses of MPN patient samples FACS sorted into stem and progenitor populations, we have found a cell type and context specific nature of these enzymes, notably the upregulation of APOBEC3C (A3C) in the high-risk Myelofibrosis (MF) stem cell population as compared to normal aged counterparts. Through lentiviral overexpression of each APOBEC3 enzyme, we can now study the effects of changes in APOBEC3 transcript level in relation to the known changes in expression seen in many cancers, focusing on the upregulation of A3C. Using these techniques, we have identified novel RNA and DNA editing targets, as well as differential gene expression patterns of each APOBEC3 in normal CD34+ cord blood and aged normal bone marrow. Gene set enrichment analysis (GSEA) performed on this dataset has exposed numerous deregulated pathways brought on by exaggerated levels of APOBEC3, including changes in splicing pathways. In addition, novel identification of the relationship between A3C and ADAR1, another innate immune deaminase, has important implications in initiation and prognosis of MPNs. Both A3C and ADAR1 transcript levels are elevated in high risk MF stem cells, and co-immunoprecipitation studies reveal a direct binding of the enzymes. Furthermore, we are able to study the detailed effects of editing by both A3C and ADAR1 using editase-deficient mutant constructs, allowing for a mechanistic look into the role of these deaminases and their deregulation in vitro and in vivo using patient samples and humanized mouse models. This novel connection, as well as the role of A3C in initiation and progression of hematopoietic malignancies will continued to be studied using this system to elucidate effects on proliferation, differentiation, self-renewal, and changes to the cell cycle, in hopes of creating both a marker of early detection and potentially a druggable target.

Citation Format: Jane Marie Isquith, Adam Mark, Jessica Pham, Mary Donohoe, Luisa Ladel, Catriona Jamieson. Effects of innate immune deaminase deregulation on initiation and progression of myeloproliferative neoplasms [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2022; 2022 Apr 8-13. Philadelphia (PA): AACR; Cancer Res 2022;82(12_Suppl):Abstract nr 899.
Abstract 915: C/EBPβis required for mouse mammary tumorigenesis
Authors: Burke Caitlin M. , Rojo Michelle D. , Rosen Jeffery M. , Ackermann Tobias , Muller Christine , Calkhoven Cornelis F. , Machado Heather L.
Published: June 2022

Breast tumorigenesis relies on complex interactions between tumor cells and their surrounding environment, orchestrated by tightly regulated transcriptional networks. While much is known about the signaling pathways affecting breast cancer cell growth and apoptosis, the transcriptional programs driving these responses are less understood. C/EBPβ is a transcription factor essential for proper cellular proliferation, differentiation, tissue function, metabolism, and the immune response, and is deregulated in breast cancer. C/EBPβ and its protein isoform LIP have long been postulated to promote breast cancer progression, however, the precise mechanisms and gene regulatory networks induced by C/EBPβ are less understood. Analysis of available human patient gene expression data reveals that increased Cebpb mRNA is associated with poor overall survival amongst all breast cancer subtypes, and analysis of available liquid chromatography with tandem mass spectrometry data shows that increased C/EBPβ total protein is associated with poor overall survival in triple-negative breast cancer patients. To test the function of C/EBPβ in tumor initiation and progression, several genetic mouse models were generated. Genetic ablation of tumor C/EBPβ in mouse models of basal-like [C3(1)TAg] and luminal (MMTV-Neu) breast cancers results in a complete block in mammary tumorigenesis. Interestingly, C/EBPβ-null tumor cells upregulate genes involved in antigen presentation and interferon gamma response in vitro, while overexpression of the C/EBPβ-LIP induces chemokines involved in immune cell recruitment and immunosuppression, such as CCL5. In support, RNA sequencing of human breast cancer cells shows that deletion of Cebpb in BT-20 cells causes de-repression of immune genes. Specifically, genes associated with interferon α and γ signaling (JAK1, TNFA, CCRL16, CSF2, IL32) as well as genes involved in antigen processing and presentation (MHC class I and class II molecules) are upregulated in the absence of Cebpb. These data suggest that C/EBPβ may repress genes important for immune visibility. Ongoing studies are aimed at identifying gene regulatory programs induced by C/EBPβ and specifically by C/EBPβ-LIP. These studies will have critical implications for whether C/EBPβ-induced pathways can be targeted in combination with other drugs, such as immune checkpoint inhibitors, improving patient outcome. These studies are supported by NIH RO1CA212518 (H.L.M.).

Citation Format: Caitlin M. Burke, Michelle D. Rojo, Jeffery M. Rosen, Tobias Ackermann, Christine Muller, Cornelis F. Calkhoven, Heather L. Machado. C/EBPβis required for mouse mammary tumorigenesis [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2022; 2022 Apr 8-13. Philadelphia (PA): AACR; Cancer Res 2022;82(12_Suppl):Abstract nr 915.
Abstract 927: Developing new mouse models of metastatic lung cancer
Authors: Wells Julie , Maser Richard , Tucker Ashley , McGee Teresa , Memishian Wendy , Doty Rosalinda , Bult Carol J.
Published: June 2022

INTRODUCTION: The focus of this work is to develop new mouse models for metastatic lung cancer. Lung cancer is the leading cause of cancer deaths worldwide and greater than 75% of human lung cancer deaths can be attributed to metastasis. Metastatic disease in most existing mouse models of pulmonary adenocarcinoma is typically sporadic and often requires aging mice for several months. These factors limit the usefulness of mouse models for basic and pre-clinical research aimed at identifying effective treatments of metastatic disease.

METHODS AND RESULTS: We have developed new mouse models of pulmonary adenocarcinoma in which we added mutations in Dicer1, an RNAse III enzyme within the microRNA (miRNA) biosynthesis pathway to a mouse model of Kras-driven pulmonary adenocarcinoma. For both models, we induced tumorigenesis by conditionally expressing an oncogenic allele of Kras (KrasG12D), deleting of both alleles of Trp53 and deleting one allele of Dicer1 in one cell population then expressing truncated Dicer1 in a different cell population. In the absence of Dicer1 truncation, mice expressing KrasG12D and deleting Trp53 in Club cells have a reported median survival of 28.6 weeks after tumor induction. When we also deleted one allele of Dicer1 in Club cells and truncated Dicer1 in alveolar type II cells of these mice, they exhibited respiratory distress and began dying 12.1 weeks after tumor induction. In our initial pilot study, all animals (12/12) developed pulmonary adenocarcinoma and 30% (3/10) had lymph node metastasis. We did not observe accelerated tumorigenesis, however, when we switched cell types, inducing tumorigenesis in alveolar type II cells and truncating Dicer1 in Club cells, or when induced tumorigenesis and truncating Dicer1 within the same cell populations.

CONCLUSIONS: Through cell type specific truncation/deletion of Dicer1 we have generated a new mouse model that rapidly develops pulmonary adenocarcinomas and metastatic disease. Our findings support our hypothesis that tumorigenesis and metastasis are influenced by miRNA regulated communication between different cell types. These models have potential for both understanding the basic processes of metastasis and for pre-clinical studies aimed at preventing and/or treating metastatic lung cancer.

Citation Format: Julie Wells, Richard Maser, Ashley Tucker, Teresa McGee, Wendy Memishian, Rosalinda Doty, Carol J. Bult. Developing new mouse models of metastatic lung cancer [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2022; 2022 Apr 8-13. Philadelphia (PA): AACR; Cancer Res 2022;82(12_Suppl):Abstract nr 927.
Abstract 945: Gab2 deficiency protects against Flt3-ITD driven acute myeloid leukemia in vivo
Authors: Spohr Corinna , Poggio Teresa , Andrieux Geoffroy , Schönberger Katharina , Cabezas-Wallscheid Nina , Börries Melanie , Halbach Sebastian , Illert Anna L. , Brummer Tilman
Published: June 2022

Internal tandem duplications (ITD) of the FMS-like tyrosine kinase 3 (FLT3) frequently co-exist with inactivating DNMT3A mutations and predict poor prognosis in acute myeloid leukemia (AML). In vitro studies implicated the docking protein Grb2-associated binder 2 (GAB2) as FLT3-ITD effector. By utilizing a Flt3-ITD knock-in, Dnmt3a haploinsufficient mouse model, we demonstrate that Gab2 is essential for the development of Flt3-ITD driven AML in vivo, as Gab2 deficient mice survived significantly longer, had reduced blast counts and exhibited an attenuated spleen and liver pathology as compared to their Gab2-proficient littermates. Further, leukemic bone marrow isolated from Gab2 deficient mice displayed a reduced colony forming unit capacity and an increased sensitivity towards the Flt3 inhibitor quizartinib. Using RNA sequencing, we identify the genes encoding for Axl and the Ret co-receptor Gfra2 as targets of the Flt3-ITD/Gab2/Stat5 axis. We propose a pathomechanism in which Gab2 increases signaling of these receptors as it induces their expression and additionally serves as a downstream effector. Consequently, Gab2 promotes AML aggressiveness as well as drug resistance by incorporating the receptor tyrosine kinases Axl and Ret into the Flt3-ITD signaling network. Therefore, our data identify GAB2 as a promising biomarker as well as a therapeutic target in human AML.

Citation Format: Corinna Spohr, Teresa Poggio, Geoffroy Andrieux, Katharina Schönberger, Nina Cabezas-Wallscheid, Melanie Börries, Sebastian Halbach, Anna L. Illert, Tilman Brummer. Gab2 deficiency protects against Flt3-ITD driven acute myeloid leukemia in vivo [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2022; 2022 Apr 8-13. Philadelphia (PA): AACR; Cancer Res 2022;82(12_Suppl):Abstract nr 945.
Abstract 974: Genome-wide analysis of estrogen receptor b binding sites: Implications for endogenous anti-metastatic activity in inflammatory breast cancer
Authors: Karagounis Ilias V. , Tastsoglou Spyros , Karavangeli Anna , Maity Amit , Chatzigeorgiou Artemis , Thomas Christoforos
Published: June 2022

Inflammatory breast cancer (IBC) is a highly metastatic breast carcinoma with high frequency of estrogen receptor a (ERa) negativity. We previously explored the second ER subtype, (ERb) and reported expression in IBC tumors and its correlation with reduced metastasis in patients. We have also seen decreased metastasis of IBC tumors when ERb is expressed or activated by chemical agonists. In contrast, ablation of ERb in IBC cells increased migration and stimulated signaling pathways that activate Rho GTPases promoting actin reorganization. We have now mapped all ERb binding sites in IBC cells and by interrogating this information together with gene expression data we identified genomic targets that mediate its tumor repressive effects. Through a combined analysis of cells that express endogenous ERb and knockout cells engineered to re-express the receptor we have defined how a biologically relevant ERb interacts with the cancer cell genome to control metastatic signals in IBC. We reveal key binding motifs and cis-regulatory chromatin sites and associate target genes with clinical outcomes to strengthen the relevance of ERb signaling for the progression of IBC. Our findings thus offer an opportunity to better understand the mechanism of ERb action that is critical step in validating its tumor repressive role in breast cancer.

Citation Format: Ilias V. Karagounis, Spyros Tastsoglou, Anna Karavangeli, Amit Maity, Artemis Chatzigeorgiou, Christoforos Thomas. Genome-wide analysis of estrogen receptor b binding sites: Implications for endogenous anti-metastatic activity in inflammatory breast cancer [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2022; 2022 Apr 8-13. Philadelphia (PA): AACR; Cancer Res 2022;82(12_Suppl):Abstract nr 974.
Abstract 99: Interplay between CDK5 and IRF2BP1 enhances breast cancer brain metastasis
Authors: Yuzhalin Arseniy E. , Lowery Frank , Saito Yohei , Zhang Patrick , Yu Dihua
Published: June 2022

Prognosis of patients with breast cancer brain metastasis is poor due to unavailability of effective treatments. Hence, more fundamental research is needed to broaden our understanding of mechanisms underlying the origin and development of brain metastasis. We have identified that expression of cyclin-dependent kinase 5 (CDK5) in cancer cells promotes brain metastasis growth in experimental mouse models through suppressing the antitumor immunity. Here, we interrogated the underlying mechanism that connects CDK5 with defective antitumor immunity. To understand the repertoire of CDK5 phosphorylation targets, we performed a quantitative stable isotope labeling by amino acids in cell culture (SILAC) phopshoproteomics analysis of cultured brain-seeking cell lines that have been genetically engineered to express shRNA targeting the CDK5 gene. The knockdown of CDK5 resulted in a dramatic dephosphorylation of multiple proteins, among which Interferon Regulatory Factor 2 Binding Protein 1 (Irf2bp1) is well recognized to function in mediating the immune response. Knockdown of CDK5 reduced the phosphorylation of Irf2bp1 at Ser66 in both 4T1 and T11 cells (Log10 fold change: -0.51 and -0.39, respectively, P less than 0.0001). These findings were further validated by co-immunoprecipitation of Irf2bp1 in cell lysates from shCtl and shCDK5 brain-seeking cancer cell lines, followed by immunoblotting for phosphoserine. Furthermore, analysis of Irf2bp1 protein band shifts and intensities using Phos-tag™ gels also demonstrated that CDK5 knockdown reduces the Irf2bp1 phosphorylation. In order to investigate the function of Irf2bp1, we established a stable knockdown of the Irf2bp1 gene in brain metastasis-derived cell lines 4T1.Br3 and T11.Br1. We observed that Irf2bp1-knockdown cells displayed a reduced expression of genes encoding the class I major histocompatibility complex (MHC-I) as well as Nlrc5, a key transcriptional regulator of MHC I. Thus, these findings indicate that CDK5 may regulate the expression of surface MHC-I in breast cancer brain metastasis cells through Irf2bp1 phosphorylation at Ser66, in the absence of Interferon gamma or other classically recognized MHC-I agonists. These findings reveal a previously unknown mechanism of MHC-I regulation in cancer cells, that might improve our knowledge about cancer cell immune evasion through MHC-I loss.

Citation Format: Arseniy E. Yuzhalin, Frank Lowery, Yohei Saito, Patrick Zhang, Dihua Yu. Interplay between CDK5 and IRF2BP1 enhances breast cancer brain metastasis [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2022; 2022 Apr 8-13. Philadelphia (PA): AACR; Cancer Res 2022;82(12_Suppl):Abstract nr 99.
Abstract CT161: GP2 immune response a predictor of recurrence in a phase IIb study evaluating HER2/neu peptide GP2 (GLSI-100) vs. GM-CSF alone after adjuvant trastuzumab in HER2 positive women with breast cancer
Authors: Patel Snehal S. , McWilliams David B. , Patel Mira S. , Thompson Jaye , Daugherty F. Joseph
Published: June 2022

The authors did not submit an updated abstract. The original abstract should be considered final.

Citation Format: Snehal S. Patel, David B. McWilliams, Mira S. Patel, Jaye Thompson, F. Joseph Daugherty. GP2 immune response a predictor of recurrence in a phase IIb study evaluating HER2/neu peptide GP2 (GLSI-100) vs. GM-CSF alone after adjuvant trastuzumab in HER2 positive women with breast cancer [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2022; 2022 Apr 8-13. Philadelphia (PA): AACR; Cancer Res 2022;82(12_Suppl):Abstract nr CT161.
Abstract LB018: Clonal evolution and lineage choice driven by IRF4 in zebrafish T-cell lymphoma model
Authors: Amanda Stella , Tan Tze King , Ong Jolynn Zu Lin , Theardy Madelaine Skolastika , Wong Regina Wan Ju , Huang Xiao Zi , Ali Muhammad Zulfaqar , Yan Li , Gong Zhiyuan , Inagaki Hiroshi , Foo Ee Yong , Pang Brendan , Tan Soo Yong , Iida Shinsuke , Sanda Takaomi
Published: June 2022

IRF4 is a master regulator of immunity and is also frequently overexpressed in mature lymphoid neoplasms. Here, we demonstrate the stage-specific oncogenicity of IRF4 in vivo, its potential effects on T-cell development and clonal evolution using a zebrafish model. IRF4-transgenic zebrafish develop aggressive tumors with massive infiltration of abnormal lymphocytes that spread to distal organs. Many late-stage tumors are mono- or oligoclonal, and tumor cells can expand in recipient animals after transplantation, demonstrating their malignancy. Mutation of p53 significantly accelerates tumor onset, increases penetrance, and results in tumor heterogeneity. Unexpectedly, single-cell RNA-sequencing reveals that majority of tumor cells are double-negative T-cells, many of which express tcr-γ that became dominant as the tumors progress, whereas double-positive T-cells are largely diminished. Gene expression and enhancer profiling demonstrates that gata3, mycb and several genes that were previously not implicated in cancers including lrrn1, patl1 and psip1 are specifically upregulated in tumors, while genes responsible for T-cell differentiation including cxcr4b, id3 and cd8a are repressed. IRF4-driven tumors are sensitive to treatment with the BRD inhibitor (JQ1).

Citation Format: Stella Amanda, Tze King Tan, Jolynn Zu Lin Ong, Madelaine Skolastika Theardy, Regina Wan Ju Wong, Xiao Zi Huang, Muhammad Zulfaqar Ali, Li Yan, Zhiyuan Gong, Hiroshi Inagaki, Ee Yong Foo, Brendan Pang, Soo Yong Tan, Shinsuke Iida, Takaomi Sanda. Clonal evolution and lineage choice driven by IRF4 in zebrafish T-cell lymphoma model [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2022; 2022 Apr 8-13. Philadelphia (PA): AACR; Cancer Res 2022;82(12_Suppl):Abstract nr LB018.