bluuebunny/crossref_metadata_2025 huggingface dataset:
Abstract 6009: Developing drug-induced resistant tumor models for efficacy evaluation of next-generation anticancer therapies
Authors: Ni Ting , Liu Gaoxiang , Huo Tongxin , Shi Wenting , Gu Qingyang , Ji Qunsheng
Published: June 2022

The therapeutic landscape of cancer has been transformed over the last few decades. Our understanding of cancer and its therapeutic approach has improved greatly due to advanced cancer biology, functional imaging and next-generation sequencing. One of the key challenges in cancer therapy is how to effectively kill cancer cells while leaving normal cells intact. As specific oncogenic drivers have been successfully identified, targeted treatments have achieved great success in clinical practice. However, acquired drug resistance remains the major barrier to sustained success of targeted cancer therapy. Those patients who initially respond well to targeted therapies eventually become resistant to the treatments. Uncovering the underlying mechanism would help develop novel therapies that overcome the drug resistance.To mimic the clinical resistance after long-term drug treatment, we established a panel of drug-induced resistant tumor models by continuous dosing of targeted drugs to tumor-bearing mice or cancer cell lines, covering a series of first-line targeted drugs including Sotorasib, Palbociclib, Ibrutinib, Capmatinib, Fulvestrant, Tamoxifen and TDM-1. Resistant models were derived from sensitive ones harboring specific oncogenic drivers. After treated with targeted drugs and passaged several times, resistant models exhibited stable resistance phenotype. Given that target gene mutation, alternative pathway activation or tumor microenvironment evolution could promote therapy resistance and cancer progression, we performed transcript and protein analysis to explore the molecular mechanism, and evaluated the potential therapeutic strategies overcoming drug resistance in the established models. Taken together, drug-induced resistant tumor models provide a promising opportunity to the better understanding of drug resistance mechanisms and to the accelerated development of next-generation anticancer agents.

Citation Format: Ting Ni, Gaoxiang Liu, Tongxin Huo, Wenting Shi, Qingyang Gu, Qunsheng Ji. Developing drug-induced resistant tumor models for efficacy evaluation of next-generation anticancer therapies [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 6009.
Abstract 6018: Expansion of mouse mammary epithelial stem cells in serum-free organoid growth medium
Authors: Rowbotham David , Rothwell Samantha , Eaves Allen C. , Louis Sharon A. , Stingl John
Published: June 2022

Growing mammary epithelial cells as organoids in a three-dimensional (3D) cell culture environment represents a more physiological model system than conventional 2D adherent cell culture systems for studying many different aspects of mammary epithelial cell biology. We have developed MammoCult™ Organoid Growth Medium (Mouse), a serum- and phenol red-free medium for the long-term propagation of organoids from mouse mammary tissue. To initiate the cultures, mouse mammary glands are resected and enzymatically dissociated sequentially in Gentle Collagenase/Hyaluronidase, trypsin, and dispase/DNase I to generate a single-cell suspension, and 4.5 x 103 of the liberated cells are then embedded in Corning® Matrigel® and maintained in MammoCult™ Organoid Growth Medium (Mouse). Alternatively, freshly dissociated mammary cells can be expanded in adherent culture in EpiCult™ Plus Medium prior to seeding into Matrigel®. Approximately 8.4 ± 0.6% (mean ± SEM; n=10) of the freshly dissociated cells seeded in the Corning® Matrigel® will proliferate and generate organoids that are typically > 200 µm in diameter and have a highly branched morphology, although organoids with a cystic morphology are also observed. Immunostaining of these branched organoids reveals that they are composed of a polarized epithelium made up of keratin (K) 8-expressing luminal cells and K5/14-expressing basal cells (n=7). Organoids upregulate transcripts for caseins (Csn1s1, Csn2), whey acidic protein (Wap), and lactoferrin (Ltf) upon stimulation with prolactin, hydrocortisone, and insulin (n=4). These multilineage organoids can be established from multiple inbred mouse strains, including C57Bl6/J, FVB, BALB/c, and CD-1. Organoids can be passaged every 10 - 14 days as small fragments at a 1:3 to 1:4 split ratio, and can be maintained for a minimum of 10 passages while still maintaining multilineage potential and a normal karyotype. These results demonstrate that MammoCult™ Organoid Growth Medium (Mouse) efficiently generates and expands mammary epithelial cell-derived organoids and offers researchers a novel tool for in vitro mammary studies.

Citation Format: David Rowbotham, Samantha Rothwell, Allen C. Eaves, Sharon A. Louis, John Stingl. Expansion of mouse mammary epithelial stem cells in serum-free organoid growth medium [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 6018.
Abstract 6027: Layered 3D co-cultured tumor model including vascularized stromal tissue may reflect drug sensitivities in vivo tumor
Authors: Takahashi Yuki , Kitano Shiro , Shinozaki Eiji , Nagayama Satoshi , Katayama Ryohei , Fujita Naoya
Published: June 2022

Background: Multiple 3D culture models have been reported as a superior tumor model than 2D culture. The importance of interaction between cancer and stromal cells has been widely recognized in tumor progression and resistance to treatment. Therefore, ex vivo model recapturing the tumor microenvironment is needed to evaluate the drug efficacy under the condition imitating the patient tumor tissue. Purpose: We developed a unique 3D co-cultured tumor model with stromal tissues containing a microvascular network. Here, we investigated drug sensitivity in conventional 2D culture, our 3D co-cultured model and in vivo tumor.

Methods: Drug sensitivity and gene expression on our model were evaluated using patient-derived cancer cells (PDC) established from colorectal cancer (CRC) patients in JFCR. The characteristics were compared with those of conventional 2D cultured cells or patient-derived xenograft (PDX).

Results: In our 3D model, the drug sensitivities tended to be decreased in comparison with those of 2D. However, some drugs presented potent efficacy in our 3D model rather than 2D, and such drugs showed significant tumor growth suppression or tumor regression in vivo. Transcriptome profiles of our 3D model showed relatively higher similarity to those of in vivo tumors than those of 2D.

Conclusion: Our study proposed the unique 3D co-cultured tumor model which may enable more accurate evaluation of drug sensitivities reflecting the in vivo circumstances.

Citation Format: Yuki Takahashi, Shiro Kitano, Eiji Shinozaki, Satoshi Nagayama, Ryohei Katayama, Naoya Fujita. Layered 3D co-cultured tumor model including vascularized stromal tissue may reflect drug sensitivities in vivo tumor [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 6027.
Abstract 603: Discovery of CD200R1 as a novel immuno-oncology target using pleiotropic signals from 23andMe’s genetic and health survey database
Authors: Fang Xin , Chung Wei-Jen , Fenaux Jill , Chen Alice , Batada Nizar , Schmidt Maike , Majeed Sophia R. , Kumar Sushil , Pitts Steven
Published: June 2022

Using large genetic and phenotypic datasets, 23andMe has developed a novel immuno-oncology target-identification platform. 23andMe has assembled a genetic database of nearly 12 million individuals, with 80 percent consented to research, coupled with health survey data comprising billions of responses. We identified hundreds of shared GWAS signals across phenotypes that appear to be driven by pleiotropic causal variants whose direction of effect (risk versus protection) systematically differs across cancer and autoimmune diseases including a signal linked to a clinically validated immune checkpoint receptor. This observation suggests that genes linked to such signals may be utilized by tumor cells to evade the immune system. We hypothesize that such signals indicate nodes of critical importance for anti-tumor immune response. By utilizing expression quantitative trait loci (eQTLs) and coding single nucleotide polymorphisms (SNPs) to link these signals to causal effects on genes, we identified CD200R1, an inhibitory receptor expressed on T cells and myeloid cells, as an immune checkpoint target. Immune-oncology associations were observed in multiple components of the CD200R1 pathway, including the receptor, its cognate ligand, CD200, and its downstream adaptor protein, DOK2. Expression of CD200R1 was validated in tumor-infiltrating lymphocytes using bulk and single-cell RNA-seq from various tumor types. This genetics-driven target discovery platform facilitated the development of a fully humanized monoclonal antibody that binds CD200R1 on immune cells with high affinity and enhances killing of tumor cells expressing CD200.

[X. F. and W-J. C. contributed equally to this work.]

Citation Format: Xin Fang, Wei-Jen Chung, Jill Fenaux, Alice Chen, Nizar Batada, Maike Schmidt, Sophia R. Majeed, Sushil Kumar, Steven Pitts. Discovery of CD200R1 as a novel immuno-oncology target using pleiotropic signals from 23andMe’s genetic and health survey database [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 603.
Abstract 6117: Clinical and molecular correlates of immune marker profiling and PD-L1 expression in non-small cell lung cancer
Authors: Shi Yunfei , Lei Youming , Liu Yinqiang , Duan Jin , Zhao Wei , Zhang Fujun , Lv Guoli , Wu Qian , Wu Mengmeng , Yin Jiani C. , Shao Yang
Published: June 2022

Background: Understanding the genomic landscape and tumor immune microenvironment of non-small cell lung cancer (NSCLC) may provide critical insight into the biology of tumor immune response and novel strategies for overcoming immunotherapy resistance. The association of clinical and genomic features with tumor immune microenvironment in NSCLC patients remains unclear.

Methods: In this retrospective cohort study, we profiled PD-L1 and immune cell marker expressions and performed targeted next-generation sequencing (Geneseeq Technology Inc., Nanjing) of baseline tissue samples from 100 NSCLC patients. The levels of CD8+, CD68+ and HLA-DR+ immune cells were assessed by multiplexed immunohistochemistry (mIHC) assay. Immune microenvironment markers and PD-L1 expression was analyzed for associations with clinical features and mutations at both the single gene and pathway levels.

Results: Mutations in TP53, SETD2, RET and the p53 signaling pathway were significantly associated with high PD-L1 high expression (P<0.001; P=0.002; P=0.04; P=0.001, respectively). Patients with high PD-L1 expression showed a trend towards higher stromal CD8+ lymphocytes (P=0.1), Alterations in T cell and B cell pathways were also associated with increased stromal CD8+ T cells (P=0.03 and P=0.08, respectively). The infiltration of intratumoral CD68(+)HLA-DR(+) M1-like macrophages positively correlated with high PD-L1 high expression (P=0.07), Hippo pathway alterations (P=0.06). By contrast, the levels of intratumoral M1-like macrophages negatively correlated with alterations in the NRF2/KEAP1 pathway (P=0.005), which might account for the poor immunotherapy outcome in this subset of patients.

Conclusions: Our study demonstrates the complex relationship between gene mutations and aberrations in signaling pathways with the immune microenvironment in NSCLC. Specific molecular features are associated with unique immune profiles and may impact the response to immune checkpoint inhibitors.

Citation Format: Yunfei Shi, Youming Lei, Yinqiang Liu, Jin Duan, Wei Zhao, Fujun Zhang, Guoli Lv, Qian Wu, Mengmeng Wu, Jiani C. Yin, Yang Shao. Clinical and molecular correlates of immune marker profiling and PD-L1 expression in 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 6117.
Abstract 6118: Tumor-derived GDF-15 promotes immune escape of tumors by functional alteration of the myeloid compartment
Authors: Haake Markus , Haack Beatrice , Genßler Sabrina , Weigandt Julia , Auer Marlene , Thiemann Vincent , Haq Wahid M. , Haag Melanie , Wedekink Florian , Fischer Birgitt , Klar Kathrin , Wölfl Matthias , Schuberth-Wagner Christine , Wischhusen Jorg
Published: June 2022

Background: Growth and differentiation factor 15 (GDF-15), a divergent member of the TGF-β protein superfamily, shows low physiological baseline expression. GDF-15 is, however, strongly upregulated during pregnancy. It is further induced in stressed and damaged tissues, where it limits immune infiltration and inflammation. In solid tumors, GDF-15 was shown to be a key inhibitor of T-cell infiltration. While this might already explain the strong correlation between GDF-15 overexpression and poor survival with or without checkpoint-based immunotherapy, effects of GDF-15 on dendritic cells and monocytes may further shape the tumor microenvironment.

Methods: To analyze the impact of GDF-15 in murine tumor models, GDF-15 was either deleted by CRISPR/CAS9 gene editing or neutralized by administration of a blocking antibody. Effects on tumor growth were recorded and the composition of the tumor microenvironment was characterized by flow cytometry. More detailed insight into tumor-immune interactions was obtained via the CrownBio Mouse I/O RNA-Seq Panel. Specific effects on polarization of innate and on antigen-specific priming of adaptive immune cells were confirmed in cellular assays in vitro.

Results: Tumor-derived GDF-15 modulates the tumor microenvironment by inhibiting infiltration and activation of myeloid cells. GDF-15 thereby impairs the induction of antitumoral immune responses. Deletion of GDF-15 enhances infiltration of innate cells into immune-excluded tumors, supports the priming of naïve T cells by dendritic cells and generates a pro-inflammatory tumor microenvironment. In vitro, GDF-15 inhibits DC maturation and synapse formation, thus preventing successful T-cell activation. Moreover, GDF-15 interferes with M1 polarization of macrophages.

Conclusion: GDF-15 secretion helps tumors to generate a microenvironment that is poorly infiltrated by immune cells. GDF-15 further polarizes myeloid cells towards a tumor-promoting, anti-inflammatory phenotype. By inducing a more pro-inflammatory tumor phenotype, anti-GDF-15 antibodies may synergize with other immunotherapeutic agents. A clinical trial combining anti-GDF-15 (CTL002) with anti-PD-1 (NCT04725474) is ongoing.

Citation Format: Markus Haake, Beatrice Haack, Sabrina Genßler, Julia Weigandt, Marlene Auer, Vincent Thiemann, Wahid M. Haq, Melanie Haag, Florian Wedekink, Birgitt Fischer, Kathrin Klar, Matthias Wölfl, Christine Schuberth-Wagner, Jorg Wischhusen. Tumor-derived GDF-15 promotes immune escape of tumors by functional alteration of the myeloid compartment [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 6118.
Abstract 6129: Investigate the mechanism and treatment efficacy of regorafenib combined with anti-PD-L1 on bladder cancer
Authors: Pai Kai-Jen , Chang Yuan , Chiang I-Tsang , Hsu Fei-Ting
Published: June 2022

Bladder cancer is one of the most common cancer in urinary system in Taiwan. According to Ministry of Health and Welfare’s research, bladder cancer plays 15th leading causes of death in Taiwan in 2020. The patients who suffer from bladder cancer after undergoing resection surgery about 60% will relapse, and 80% of them will relapse in less than a year, 15% of the relapsing patients will aggravate and become invasive carcinoma spread all over the body. PD-L1 (Programmed death-ligand 1), also known as B7-H1 and CD274 is a transmembrane glycoprotein, which interacts with PD-1 (Programmed death-1) receptor on T cells to elicit the immune checkpoint response. Overexpression of PD-L1 suppresses the function of CD8+ T cells and facilitate tumor progression. Targeting tumor PD-L1 may be a possible treatment option; however, PD-L1 expression was only detected in small amount of population. Regorafenib is a multiple tyrosine kinase inhibitor, which has been approved by the US Food and Drug Administration to treat advanced HCC. In our research, we found that regorafenib may upregulation the surface PD-L1 expression in both mice and human cell line. Based on this finding, we suggested that regorafenib may show potential to enhance treatment efficacy anti-PD-L1. Therefore, we established MB-49 bearing animal model to identify the immune regulation and treatment efficacy of regorafenib combined with anti-PD-L1. In ours animal data we found that combination therapy effectively increased the tumor infiltrating population of NK cells and CD8+ lymphocytes which were determined by flow cytometry assay. Not only anti-tumoral leukocytes, we also detected the population of immune suppressor cells including regulatory T cells, MDSCs and M2 macrophage was suppressed by combination therapy. We also detected the tumor microenvironment related markers (VEGF, IDO-1, PD-L1, CD8, CD31, CD49b, FOXP3, granzyme B) to evaluate combination therapy effect by using immunohistochemistry staining. In conclusion, regorafenib may effectively enhance anti-PD-L1 efficacy via triggering positive immune regulation and increasing the interaction of anti-PD-L1 with tumor.

Citation Format: Kai-Jen Pai, Yuan Chang, I-Tsang Chiang, Fei-Ting Hsu. Investigate the mechanism and treatment efficacy of regorafenib combined with anti-PD-L1 on bladder 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 6129.
Abstract 6131: Harnessing resveratrol-mediated restoration of MHC-I in melanoma to enhance immunotherapy
Authors: Morehead Lauren C. , Koss Brian S. , Miousse Isabelle R. , Tackett Alan J.
Published: June 2022

The introduction of immunotherapy has drastically improved outcomes in cancer therapy, particularly in melanoma, the deadliest form of skin cancer. However, around half of melanoma tumors are initially resistant to treatment and a significant proportion of initial responders gain resistance later. This is in part because tumors are able to decrease their ability to be recognized by T-cells, an essential actor in immunotherapeutic action. There are two major types of T-cells: CD4+ helper and CD8+ cytotoxic. The most common method used by tumor cells to evade immune attack is to down-regulate major histocompatibility complex (MHC) class I molecules, which allow CD8+ T-cells to recognize cells to be eliminated. Thus, to increase immunotherapeutic efficacy, it is critical to identify how to safely restore MHC-I expression on cancer cells so that tumor antigens are presented to T-cells. I focused on finding potential adjuvants to immunotherapy for this purpose, and I found that resveratrol, a natural compound found in plants and associated with various medicinal properties, consistently elevated MHC-I expression in melanoma cells. Through a proteomics study, I identified that resveratrol also significantly affected pathways known to enhance MHC-I expression.

Citation Format: Lauren C. Morehead, Brian S. Koss, Isabelle R. Miousse, Alan J. Tackett. Harnessing resveratrol-mediated restoration of MHC-I in melanoma to enhance immunotherapy [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 6131.
Abstract 6142: Gain-of-function p53R172H mutation drives accumulation of neutrophils in the pancreatic tumor microenvironment promoting resistance to immunotherapy
Authors: Siolas Despina , Vucic Emily , Kurz Emma , Hajdu Cristina , Bar-Sagi Dafna
Published: June 2022

The immune microenvironment plays a critical role in cancer development, progression and resistance to therapy. Specific tumor genotypes can profoundly influence the composition of the immune microenvironment. After oncogenic KRAS, TP53 is the most frequently mutated gene in pancreatic ductal adenocarcinoma (PDAC). KRASG12D mutations in tumor cells are known to modify the immune landscape of PDAC, however the immune effects of neomorphic TP53 mutations have not been defined. Focusing on the most common neomorphic TP53 mutation in human PDAC, we sought to evaluate the non-cell-autonomous role of gain-of-function mutant Trp53R172H in modulating the immune microenvironment of pancreatic tumors by using KrasG12D-mutated mouse pancreatic ductal epithelial cells (PDEC) genetically engineered to express the Trp53R172H gain-of-function. Orthotopically implanted tumors derived from CRISPR-generated KrasG12D/+;Trp53R172H/+ PDEC had a distinct immune profile in comparison to KrasG12D/+;Trp53+/+ tumors, characterized by an influx of intratumoral CD11b+Ly6G+ neutrophils and concomitant decreases in CD3+ T cells, CD8+ T cells, and CD4+ T helper 1 (Th1) cells. Analysis of publicly available human PDAC cohorts revealed enrichment of genes in neutrophil-related pathways in TP53-mutated tumors. Knockdown of CXCL2, a neutrophil chemoattractant, in the tumor epithelial compartment of CRISPR KrasG12D/+;Trp53R172H/+ PDEC tumors reversed the neutrophil phenotype. Depleting neutrophils in mice bearing CRISPR KrasG12D/+;Trp53R172H/+ PDEC tumors augmented sensitivity to combined CD40 immunotherapy and chemotherapy. Collectively, these data link gain-of-function mutant Trp53R172H to the presence of intratumoral neutrophils in pancreatic cancer and suggests that tumor genotypes could inform patient selection for immunotherapy.

Citation Format: Despina Siolas, Emily Vucic, Emma Kurz, Cristina Hajdu, Dafna Bar-Sagi. Gain-of-function p53R172H mutation drives accumulation of neutrophils in the pancreatic tumor microenvironment promoting resistance to immunotherapy [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 6142.
Abstract 6169: Identification of molecules mediating natural killer cell-pancreatic stellate cell interactions
Authors: Malchiodi Zoe X. , Fitzgerald Allison , Galvano Elena , Weiner Louis M.
Published: June 2022

Pancreatic stellate cells (PSCs) are the main sources of cancer-associated fibroblasts in pancreatic ductal adenocarcinoma (PDAC), which account for at least 50% of PDAC’s total tumor volume and its dense desmoplastic stroma. In recent years, immune-stromal cell interactions have been the subject of research in a variety of diseases. Interestingly, natural killer (NK) cells are relatively understudied in PDAC and may be exploited to identify new treatment opportunities. We have explored how NK cell-PSC interactions are mediated by known NK cell ligands. Human PSC (hPSC, CRC-811) or PDAC (MIA-PaCa-2, PANC-1) cell lines were co-cultured with DiO stained NK cells (NK-92, donor NK cells) for 4 hours, then target cells were assessed for apoptosis by flow cytometry using AnnexinV and Sytox stains. We found that although NK cells lyse both PSCs and PDAC cells, there was more NK cell-mediated lysis against PSCs. Using flow cytometry, we found that PSCs express the well-known MICA/B ligand, which binds the activating NKG2D receptor on NK cells to initiate its cytotoxic activity. In co-culture, neutralizing MICA/B in vitro did not completely inhibit NK cell-mediated lysis of PSCs, suggesting other interactions need to be explored. To identify other molecular determinants, NK cell ligand and receptor expression was evaluated by quantitative PCR in human PSC, PDAC, NK, and other leukocytic (K562) cell lines. Cell surface validation of highly expressed NK cell ligands and receptors were confirmed by flow cytometry and/or Western blot. NK cell ligands that were both highly expressed and localized to the cell surface were: NKG2D ligands—MICA and ULBP2 (both activating), KIR2DS4 (CD158I) ligand—HLA-A (activating), and NKp44 ligands—NKp44L and PCNA (activating and inhibitory, respectively). Their corresponding receptors were also expressed and localized to the cell surface by NK cells. These results highlight novel NK cell-PSC interactions that may mediate immune suppression in PDAC. These findings warrant further investigation.

Citation Format: Zoe X. Malchiodi, Allison Fitzgerald, Elena Galvano, Louis M. Weiner. Identification of molecules mediating natural killer cell-pancreatic stellate cell interactions [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 6169.
Abstract 6206: Discerning vasculature destruction caused by novel vascular disrupting agents examined by optical and multispectral optoacoustic imaging
Authors: Wanniarachchi Hashini I. , Schuetze Regan , Arango Lorena , Hamal Khagendra , Carlson Graham J. , Pavlich Cyprian , Deng Yuling , Trawick Mary L. , Pinney Kevin G. , Liu Li , Mason Ralph P.
Published: June 2022

Tumor growth requires a functioning vascular network and offers an attractive therapeutic target. The invasive proliferating neovasculature of tumors lacks pericyte support and exhibits increased permeability thereby offering a unique, potentially selective target for anticancer therapy. Most vascular disrupting agents (VDAs) inhibit microtubule formation, causing rapid morphological changes in endothelia cells resulting in dramatically increased vessel permeability, cellular detachment, vessel occlusion, and vessel wall damage [doi: 10.3390/molecules26092551]. Highly vascular kidney tumors are expected to be particularly sensitive to VDAs. We are evaluating OXi8007, a water soluble indole phosphate pro-drug derivative of the first generation VDA combretastatin. The prodrug OXi8007 is stable in saline but releases active OXi8006 is the presence of alkaline phosphate. Preliminary pharmacokinetic data benchmarked using 13C-labeled OXi8006 and OXi8007 show rapid vascular clearance and conversion to the glucuronide conjugate in vivo. Doses up to 700 mg/kg were tolerated by mice. Luciferase-expressing RENCA tumor cells were implanted orthotopically in the right kidney capsule of syngeneic BALB/c mice. Bioluminescence Imaging (BLI) was applied weekly to assess tumor growth and acute response to OXi8007. Following addition of luciferin substrate subcutaneously BLI signal from the tumor reached a maximum within 10 minutes. When BLI was repeated 4 hrs after 250 or 350 mg/kg OXi8007 IP, the generated signal was <1% of baseline signal indicating acute vascular disruption. Multispectral Optoacoustic Tomography (MSOT) revealed matching acute changes in tumor vascular oxygenation and additionally showed distinct heterogeneity. An oxygen gas breathing challenge indicated lower oxygen saturation in the tumor following treatment. Histology showed massive hemorrhage following both doses. BLI signal recovered after 1 to 2 days and repeated administration of OXi8007 again caused acute vascular shutdown. These results match previous observations in breast tumor models [doi: 10.1016/j.canlet.2015.08.021] indicating opportunities for therapeutic application.

Citation Format: Hashini I. Wanniarachchi, Regan Schuetze, Lorena Arango, Khagendra Hamal, Graham J. Carlson, Cyprian Pavlich, Yuling Deng, Mary L. Trawick, Kevin G. Pinney, Li Liu, Ralph P. Mason. Discerning vasculature destruction caused by novel vascular disrupting agents examined by optical and multispectral optoacoustic imaging [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 6206.
Abstract 6212: Multiomics analyses reveal multimodal Fusobacterium DNA damage perturbations in colorectal cancer
Authors: Kataria Radhika , Quist Jelmar , Lee Sunjae , Megalios Anargyros , Sawyer Elinor , Hardiman Thomas , Liu Chang , Kelly Katherine , Portlock Theo , Shoaie Saeed , Grigoriadis Anita
Published: June 2022

Fusobacterium nucleatum is an opportunistic bacterial pathogen and oncogenic microbe. To systematically investigate Fusobacterium-host interactions in colorectal cancers (CRCs), using host sequencing data, we developed a multi-omics PAThogen CHaracterization tool (PATCH) allowing for gene-level annotations. In 1,020 CRC transcriptomes and 28 genomes from The Cancer Genome Atlas (TCGA), we detected 9 Fusobacterium species and genes encoding 2,346 proteins, including carcinogenic virulence factors, oxidative stress chaperones, and immune response modulators. Fusobacterium was absent in tumor-adjacent normal tissue, however showed variable intra-tumor heterogeneity. PATCH-defined Fusobacterium-positive CRCs confirmed known and revealed novel perturbations in several host DNA damage responses (DDR) mechanisms, including deficiencies in telomere maintenance pathways, elevated cytosolic DNA/RNA sensing signaling, and mutations in genes involved in multiple oncogenic-related pathways. Moreover, PATCH uncovered Fusobacterium’s DNA integrated into the coding regions of tumor suppressor genes in the cancer genome and in non- coding regions significantly enriched in close proximity to Long Interspersed Elements (LINE-1), suggesting a potential rationale for LINE-1 hypomethylation in Fusobacterium-positive CRCs. Collectively, our analytical approaches have delineated Fusobacterium’s multimodal contribution to disrupting genomic stability, which ultimately could facilitate tailored treatment options targeting several DDR pathways in Fusobacterium-positive CRC.

Citation Format: Radhika Kataria, Jelmar Quist, Sunjae Lee, Anargyros Megalios, Elinor Sawyer, Thomas Hardiman, Chang Liu, Katherine Kelly, Theo Portlock, Saeed Shoaie, Anita Grigoriadis. Multiomics analyses reveal multimodal Fusobacterium DNA damage perturbations 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 6212.
Abstract 6269: A cell double barcoding system for quantitative evaluation of primary tumors & metastasis in animals: Uncovers clonal-specific anti-cancer drug effects
Authors: Hesin Arkadi , Kumar Santosh , Gahramanov Valid , Becker Maria , Vilenchik Maria , Alexandrov Ilya , Yaglom Julia , Sherman Michael
Published: June 2022

Imaging in monitoring metastasis in mouse models have low sensitivity and is not quantitative. Cell DNA barcoding, demonstrating high sensitivity and resolution, allows monitoring effects of drugs on the number of tumor and metastatic clones. However, this technology is not suitable for comparison of sizes of metastatic clones in different animals, e.g. drug treated and untreated, due to high biological and technical variability upon tumor and metastatic growth and isolation of barcodes from tissue DNA. Yet, both numbers of clones and their sizes are critical parameters for analysis of drug effects. Here we developed a modification of the barcoding approach for monitoring drug effects on tumors and metastasis that is quantitative, highly sensitive and highly reproducible. This novel cell double barcoding system allows simultaneously following the fate of two or more cell variants or cell lines in xenograph models in vivo, and also following the fates of individual clones within each of these populations. This system allows comparing effects of drugs on different cell populations, and thus normalizing drug effects by drug-resistant lines, which corrects for both biological and technical variabilities and significantly increases the reproducibility of results. Using this barcoding system, we uncovered that effects of a novel DYRK1B kinase inhibitor FX9847 on primary tumors and metastasis is clone-dependent, while a distinct drug osimertinib demonstrated clone-independent effects on cancer cell populations. Overall cell double barcoding approach can significantly enrich our understanding of drug effects in basic research and preclinical studies.

Citation Format: Arkadi Hesin, Santosh Kumar, Valid Gahramanov, Maria Becker, Maria Vilenchik, Ilya Alexandrov, Julia Yaglom, Michael Sherman. A cell double barcoding system for quantitative evaluation of primary tumors & metastasis in animals: Uncovers clonal-specific anti-cancer drug effects [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 6269.
Abstract 627: Custom Vectra® Polaris™ fluorescent multiplex IHC panel identifies mature tertiary lymphoid structures in colorectal cancer
Authors: Pollan Sara G. , Hanifi Arezoo , Hargrove James , Parnell Erinn A. , Lin Jessica , Ragheb Josette William , Au Qingyan
Published: June 2022

Introduction: Tertiary lymphoid structures (TLS) are promising prognostic indicators of positive outcomes for patients with solid tumors including colorectal cancer (CRC) [1]. Large-scale retrospective analysis shows patients with mature TLS in particular respond to PD-1/PD-L1 antibody treatment with improved objective response, progression-free and overall survival [2]. Since not all patients respond to PD-1/PD-L1 antibody treatment, identifying patients with mature TLS is clinically relevant as it enables selection of patients likely to respond to immune checkpoint blockade. Mature TLS are composed of T cells (CD3+), follicular B cells (CD20+), germinal center B cells (CD23+), and follicular dendritic cells (CD21+) [3]. The ability to identify and evaluate TLS is limited by detection methods which traditionally employ hematoxylin and eosin (H&E) staining for visual quantification of immune aggregates, potentially significantly underestimating their quantity [4]. Multiplexed fluorescent immunohistochemistry (IHC) detection assays have the capability to precisely quantify mature TLS within the TME. Colorectal cancer specimens expressing a dynamic range of TLS are stained with a novel custom Vectra® Polaris™ multiplex immunofluorescence panel detecting CD20, CD21, CD23, CD3 and pan-cytokeratin. Identification of mature TLS with CD3+CD20+CD21+ and CD3+CD20+CD23+ expression in the TME is reported.

Results: Vectra® Polaris™ TLS multiplex immunofluorescence panel successfully identified mature TLS in the TME of colorectal cancer patient samples. TLS were quantified via custom analytics algorithms generated with Indica HALO software.

Conclusion: Characteristics of TME immunity in colorectal cancer differentially impact an individual patient’s odds of survival [5]. The novel Vectra® Polaris™ multiplex assay in this study shows a detailed picture of mature TLS in patient cancer samples. Future applications of this panel include investigations of TLS associated with successful anti-tumor immune development and therapeutic response to treatment. [1] Oncoimmunology. 2020; 9(1): 1724763 [2] Nat Cancer. 2, 794-802 (2021) [3] Oncoimmunology. 2021; 10(1): 1900508. [4] Mod Pathol 2017 Sep;30(9):1204-1212. doi: 10.1038/modpathol.2017.43. [5] Cell. 2020 Sep 3;182(5):1341-1359.e19. doi: 10.1016/j.cell.2020.07.005

Citation Format: Sara G. Pollan, Arezoo Hanifi, James Hargrove, Erinn A. Parnell, Jessica Lin, Josette William Ragheb, Qingyan Au. Custom Vectra® Polaris™ fluorescent multiplex IHC panel identifies mature tertiary lymphoid structures 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 627.
Abstract 6312: The adverse events atlas, towards a strategy to predict synergistic adverse events of combination therapies
Authors: Kucukosmanoglu Asli , Scoarta Silvia , Wijnands Thomas , Kanev George , Westerman Bart , Kiewit Bert , Noske David , Wurdinger Tom
Published: June 2022

The current gold-standard of estimating adverse events of a drug are clinical trials. However, these trials fail to represent real-life practice where patients have additional diseases (comorbidities) and commonly use numerous drugs when entering the clinic. Therefore, there is a rise of interest in combinational therapies, also because effective combinations are expected to prevent therapy resistance. At this moment it is not feasible to predict the adverse events of new combination therapies due to lack of available information both from a dimensionality (i.e., number of adverse events recorded per patient) as well as from a patient-number perspective. When available, this information allows to choose combinational therapies with acceptable adverse events.

In this study we developed a preliminary method to predict adverse events of drug combinations in order to select combinations with a mild adverse event profile.

We used the FAERS, an FDA post-marketing adverse events registry as data source containing 15 million adverse-event records. First, we developed a method to visualize the adverse events profiles of monotherapy and combination therapy using dimension reduction to accurately represent the relation between adverse events over many patients. These adverse-event profiles are then fed to a convolutional neural network (CNN) to generate an explainable prediction-model for adverse events occurring in combination therapies.

The CNN trained on monotherapy is able to learn from the data and recognize adverse event patterns. The learned pattern information, as stored in the so-called latent space, can be converted back onto the original adverse event profiles. This showed a high similarity to the original data, also for unseen combination therapy effects. Furthermore, a t-SNE analysis on the latent space of the CNN is able to separate additive and synergistic adverse event patterns in combination therapy.

Our CNN model can successfully learn complex adverse-event patterns for single drugs and their combinations, which are all encoded in the latent space. The developed method is therefore applicable to determine the combinatorial effects of highly complex adverse event profiles.

Citation Format: Asli Kucukosmanoglu, Silvia Scoarta, Thomas Wijnands, George Kanev, Bart Westerman, Bert Kiewit, David Noske, Tom Wurdinger. The adverse events atlas, towards a strategy to predict synergistic adverse events of combination therapies [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 6312.
Abstract 6393: The Cancer Genomics Cloud enables bioinformatics training in a hybrid educational world
Authors: Ray Manisha , Subramanian Sai Lakshmi , Roberson David , Worman Zelia , Dean Dennis A. , Koc Soner , DiGiovanna Jack , Davis-Dusenbery Brandi
Published: June 2022

The Cancer Genomics Cloud (CGC), powered by Seven Bridges, is an NCI-funded platform that streamlines access to large cancer datasets, bioinformatic tools, and cloud computation for cancer researchers. The attributes of the CGC designed to democratize data analysis also make it ideal for training the next generation of data scientists. During the Covid-19 pandemic, it has become clear that remote/virtual learning is of great importance for workforce development, and that reducing barriers to high quality educational resources is critical for many populations. Here we present our best practices for education of bioinformatics using the CGC, taking advantage of both distributed cloud networks and platform features to enhance learning.

Our best practices methods are focused on a systematic approach that takes the instructor and students through a typical bioinformatics workflow in their field of research. Briefly, the organizational structure of the CGC, known as “projects,'' contains all aspects of an analysis, including data files, tools, and tool settings. Projects have fine-grained permission settings, which allow the owner to securely share their project for viewing or editing. An instructor can generate an example analysis from start to finish then share an entire project with trainees. Both students and instructors have access to the same data on the cloud, so the teacher can pre-populate the projects with specific files, ensuring the same starting point. All members within the project can communicate, including adding notes directly on the results, allowing students to troubleshoot in real time with the teacher. Instructors can manage costs for the whole class through the CGC’s billing system. The CGC also provides Public Projects for self-guided training, where a researcher can learn by example using the data, tools, and completed tasks within the project, including detailed instructions embedded in markdown language.

We have successfully used these methods to train masters students in bioinformatics at Georgetown University for three years, as well as high school students, college students, and current cancer researchers.

This approach to virtual learning of bioinformatics can democratize training for the next generation of data scientists. The FAIR practices built into the CGC enables education across all levels of expertise, empowering users to drive cancer research from any stage of their career.

Citation Format: Manisha Ray, Sai Lakshmi Subramanian, David Roberson, Zelia Worman, Dennis A. Dean, Soner Koc, Jack DiGiovanna, Brandi Davis-Dusenbery. The Cancer Genomics Cloud enables bioinformatics training in a hybrid educational world [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 6393.
Abstract 6396: Detection of stemness and EMT in response to radiation in glioblastoma and ovarian cancer cells
Authors: Keniston Aaron , Enciso Alondra , Jung Yeonkyu , Antonissen Ashley , Fuller Ryan , Morcos Ann , Bertucci Antonella , Vazquez Marcello , Unternaehrer Juli
Published: June 2022

Tumor recurrence and metastasis contribute to the poor prognosis of ovarian cancer (OC) and glioblastoma multiforme (GBM). The aggressiveness of GBM and OC during recurrence creates a need to identify potential therapeutic targets. The cancer stem cell (CSC)-like phenotype and epithelial mesenchymal transition (EMT) have been shown to contribute to cancer aggressiveness, which makes CSCs and EMT viable targets for therapeutic intervention. EMT is a normal process during development and wound healing, but its reactivation in cancer cells contributes to metastasis. To detect CSC phenotypes in live cells without the use of antibodies, we are analyzing core stem cell transcription factors in which a SOX2/OCT4 response element, designated as SORE6, regulates a GFP reporter system. Also, to detect EMT we are using a GFP based reporter regulated by Zeb1 3’ UTR, a common EMT marker. Our aim is to detect changes in stemness and EMT in response to proton and photon ionizing radiation (IR). We hypothesize that proton and photon IR have similar effects on aggressiveness in OC and GBM. To investigate this, 2 GBM cancer cell lines, 1 breast cancer cell line, 1 OC cell line, and two patient derived samples were exposed to 0, 1, 2, 4 and 8Gy of 250 MeV proton and 6 MeV photon beams. 72 hours post IR exposure, flow cytometry analysis quantified reporter activity. We found an increase in stemness as radiation dosage increased in most cell lines. Results suggest that proton radiation increases stemness more than photon exposure.

Citation Format: Aaron Keniston, Alondra Enciso, Yeonkyu Jung, Ashley Antonissen, Ryan Fuller, Ann Morcos, Antonella Bertucci, Marcello Vazquez, Juli Unternaehrer. Detection of stemness and EMT in response to radiation in glioblastoma and ovarian cancer cells [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 6396.
Abstract 662: Late CTLA-4 Ig treatment improves antitumor efficacy of immunotherapy
Authors: Mok Stephen , Allison James P.
Published: June 2022

The promising approach of combining immune checkpoint therapies such as anti-CTLA-4 and anti-PD-1 increases antitumor response and overall survival rate relative to single treatments. However, it also increases the frequency and severity of immune-related adverse events (irAEs), such as cardiotoxicity. Our previous results showed that CTLA-4 Ig (abatacept), an inhibitor of T cell costimulation through CD28, can reverse those irAEs in patients with cancer, however, its effect on the antitumor response remains unclear. In the B16F10 melanoma model, we injected mice with CTLA-4 Ig antibody when: 1) mice were first treated with anti-CTLA-4, anti-PD-1, or combination (early time point) and 2) the immunotherapy was finished (late time point). We demonstrated that CTLA-4 Ig at the early time point compromised the antitumor efficacy of the immunotherapy. In contrast, the antitumor efficacy of the immunotherapy was improved if mice were treated with CTLA-4 Ig at the late time point. We also found that the antitumor response induced by CTLA-4 Ig (late) was dependent on CD80/86. Interestingly, the frequency of ICOS+ Foxp3+ Tregs was significantly reduced by CTLA-4 Ig at the late time point. Thus, we treated B16F10 tumors in Foxp3DTR mice with anti-CTLA-4 and then with CTLA-4 Ig (late). The antitumor tumor efficacy was similar between mice treated with or without Diphtheria toxin (DT) at the late time point. This suggests that the improved antitumor efficacy of CTLA-4 Ig at the late time point was Treg dependent. Collectively, we show that CTLA-4 Ig treatment has a differential role throughout the time course of immunotherapy treatments, and this work suggests a potential combination treatment strategy with checkpoint blockade and late CTLA-4 Ig treatment.

Citation Format: Stephen Mok, James P. Allison. Late CTLA-4 Ig treatment improves antitumor efficacy of immunotherapy [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 662.