bluuebunny/crossref_metadata_2025 huggingface dataset:
Abstract 4397: Chemotherapy-induced peripheral neuropathy and falls in cancer survivors relate to digital balance and gait impairments
Authors: Shah Vrutangkumar , Muzyka Daniel , Guidarelli Carolyn , Sowalsky Kristen , Winters-Stone Kerri M. , Horak Fay B.
Published: April 2023

Background and Aim: Standing postural sway and gait tests with body-worn inertial sensors provide a multitude of objective, digital balance, and gait measures that represent several different domains controlling mobility. However, it is not clear which domains of balance or gait best reflect the impact of treatment-induced peripheral neuropathy on mobility or differentiate fall risk in cancer survivors. This study aimed to determine which domains of balance and gait differed between cancer survivors who reported 1) symptoms of peripheral neuropathy versus asymptomatic survivors and 2) falls in the previous six months versus non-fallers.

Methods: Both postural sway during 30 seconds of quiet stance and gait characteristics from the Instrumented Time-Up-and-Go test (ITUG) were recorded with six synchronous inertial sensors (Opals by APDM Wearable Technology, a Clario company). The sensors were placed on both shanks and wrists, and one on the lumbar spine and sternum in 425 female cancer survivors (age: 62 ± 6 years). A principal component analysis (PCA) approach was used first to identify independent domains of mobility from 14 balance and gait measures for subsequent analysis.

Results: PCA analysis revealed 5 independent domains (PC1: Sway amplitude, PC2: Gait pace; PC3: Sway frequency; PC4: Gait spatial-temporal and PC5: Turning) that accounted for 81% of the variance of performance across the participants. Cancer survivors who reported neuropathy showed a significantly higher sway frequency (PC3) than asymptomatic survivors. Cancer survivors who reported falls showed a significantly larger sway area (PC1) and slower gait pace (PC2) than non-fallers.

Conclusions: Wearable sensor instrumentation from short balance and gait tests capture objective digital gait and balance outcomes reflective of treatment-related neuropathy and fall risk in women treated with chemotherapy for cancer. Wearable sensors could be used for oncology clinical trials to assess the impact of cancer treatment and symptom mitigation strategies to limit changes in mobility that might lead to excess falls in people with cancer.

Citation Format: Vrutangkumar Shah, Daniel Muzyka, Carolyn Guidarelli, Kristen Sowalsky, Kerri M. Winters-Stone, Fay B. Horak. Chemotherapy-induced peripheral neuropathy and falls in cancer survivors relate to digital balance and gait impairments. [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2023; Part 1 (Regular and Invited Abstracts); 2023 Apr 14-19; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2023;83(7_Suppl):Abstract nr 4397.
Abstract 4401: The precision chemotherapy to improve the epithelial ovarian cancer prognosis
Authors: Pak Seongeun , Choi Youn Jin
Published: April 2023

Purpose: Recurrence is common in ovarian cancer patients, even in those patients who show complete response after first-line chemotherapy, 80% of the patients experience a recurrence. So gynecologic oncologists often have to consider several factors when choosing next chemotherapy regimen. Therefore, this study aimed to find serial chemotherapy regimen choice to anticipate patients’ better prognosis.

Materials and Methods: We reviewed 656 patients diagnosed ovary cancer in one tertiary institutional hospital between 2006 and 2020. Patents’ clinic-pathologic characteristics, primary treatment, chemotherapy regimens, survival outcomes were collected. We divided patients into several groups according to their chemotherapy regimen by mechanism of action and compared them. 1st line chemotherapy(CTx) type 1: carboplatin-paclitaxel, carboplatin-docetaxol, cisplatin-paclitaxel, cisplatin-docetaxol2nd line CTx type 1: carboplatin-paclitaxel, carboplatin-docetaxol, cisplatin-paclitaxel, cisplatin-docetaxol2nd line CTx type 4: carboplatin-gemcitabine2nd maintenance CTx type 2: PARP inhibitor(PARPi)2nd line CTx Type 5: Platinum + topoisomerase inhibitor 2nd line CTx Type 6(cisplatin-vinorelbine)2nd line CTx Type 8(topoisomerase inhibitor only) 2nd line CTx Type12 (single taxol) The survival rate of ovary cancer patients decreased from stage 1 to stage 4.Among 1st line CTx type 1 and 2nd line CTx type(1 or 4) patients, those who used PARPi in 2nd line maintenance CTx(n=14) had a higher death rate than those who did not receive maintenance CTx or received that other than PARPi(hazard ratio 1.44). In the 1st line CTx 1~4 group, we divided patients according to 2nd line CTx type. We compare 2nd line CTx type 5 group(n=30) with type 6+8+12 group(n=26). The death rate was of type 5 group was higher than type 6+8+12 group. And Overall survival of 2nd line CTx type 5 group was 22 months, that of 2nd line CTx type 6+8+12 group was 26 months.

Conclusion: When changing the chemotherapy regimen in the recurrence of ovarian cancer, the survival rate or overall survival may vary depending on the order in which the regimen is used. If we collect patients data more broadly, the predictive power will be better.

Citation Format: Seongeun Pak, Youn Jin Choi. The precision chemotherapy to improve the epithelial ovarian cancer prognosis. [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2023; Part 1 (Regular and Invited Abstracts); 2023 Apr 14-19; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2023;83(7_Suppl):Abstract nr 4401.
Abstract 4403: The Efficacy of Immune Checkpoint Inhibitors in Advanced Biliary Tract Cancer with KRAS Mutation
Authors: Jeong Sun Young , Jang Jaeyeon , Jeon Youngkyung , Jung Ye Ji , Choi Daeho , Hong Joohyun , Kim Seung Tae , Hong Jung Yong , Park Joon Oh , Park Young Suk , Lim Ho Yeong
Published: April 2023

BackgroundWith a 15% of incidence, KRAS is one of the most common mutations in biliary tract cancer (BTC) and a poor prognostic factor. In BTC, the immune checkpoint inhibitor (ICI) as salvage therapy has modest activity. There was little data on the efficacy of ICIs according to the status of KRAS mutation in BTC.

MethodWe conducted molecular profiling in BTC patients who received the ICIs as salvage therapy. Programmed death ligand 1 (PD-L1) expression on tumor cells and tumor-infiltrating lymphocytes (TIL) was also assessed. We analyzed overall survival (OS) and progression-free survival (PFS) of ICI in BTC patients according to the status of PD-L1 expression and KRAS mutation.

ResultA total of 62 patients were included in this analysis. The median age was 68.0. 47 patients (75.8%) received pembrolizumab and 15 (24.2%) received nivolumab as salvage therapy. All patients received gemcitabine plus cisplatin as the frontline therapy, and 53.2% had fluoropyrimidine plus oxaliplatin (FOLFOX) before ICIs. The median lines of prior chemotherapy were 2.5. KRAS mutation was found in 14 patients (22.6%). 28 patients (45.2%) were positive for PD-L1 CPS. There was no statistical correlation between the status of KRAS mutation and PD-L1 expression. The median OS and PFS to ICI were 5.6 (IQR; 3.3 - 8.0) and 3.8 (IQR; 3.0 - 4.5) months, respectively. There were no statistically significant differences in PFS to ICIs according to the status of KRAS mutation (Mutant type vs. Wild type) and PD-L1 expression (Positive vs Negative). In subgroup analysis, patients with both KRAS mutation and PD-L1 positivity had longer PFS as compared to patients with KRAS mutation and PD-L1 negativity (6.5 vs 2.6 months, p=0.047). This finding was not shown in patients with a wild type of KRAS.

ConclusionThis analysis showed that PD-L1 expression might be a novel biomarker for ICIs in BTC patients with KRAS mutation but not the wild type of KRAS.

Citation Format: Sun Young Jeong, Jaeyeon Jang, Youngkyung Jeon, Ye Ji Jung, Daeho Choi, Joohyun Hong, Seung Tae Kim, Jung Yong Hong, Joon Oh Park, Young Suk Park, Ho Yeong Lim. The Efficacy of Immune Checkpoint Inhibitors in Advanced Biliary Tract Cancer with KRAS Mutation. [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2023; Part 1 (Regular and Invited Abstracts); 2023 Apr 14-19; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2023;83(7_Suppl):Abstract nr 4403.
Abstract 4433: Galectin-3 synergizes with CD47 to suppress phagocytosis and T cell immunity in peritoneal metastases of gastric adenocarcinoma
Authors: Fan Yibo , Song Shumei , Li Yuan , Dhar Shilpa , Jin Jiankang , Yao Xiaodao , Wang Ruiping , Wang Ailing , Pizzi Melissa Pool , Wu Jingjing , Yoshimura Katsuhiro , Ma Lang , Hanash Samir , Calin George , Wang Linghua , Curran Michael , Ajani Jaffer
Published: April 2023

Peritoneal metastasis (PC), the common sites of gastric adenocarcinoma (GAC) metastasis, remains resistant to therapies available in the clinics and new target is strived to be found. CD47 is one of the major players in conveying the “don’t eat me” signal upon binding its receptor SIRPα on myeloid cells which evade innate immune responses. Previously clinical studies on the blockade of CD47 alone resulted in limited clinical benefits suggesting that other target(s) must be inhibited simultaneously with CD47 for gaining advantage. Here, we found that CD47 was highly expressed on peritoneal malignant cells and its overexpression was a poor prognosticator. Additionally, we observed that Galectin-3 (Gal-3) was highly enriched in tumor cluster and was positively correlated with CD47 expression in short survival of patients revealed by scRNA-seq and metastatic GAC samples. Moreover, we found double positive of Gal-3 and CD47 significantly associated with diffuse type, poor differentiation, and tumor relapse. Mechanistically, Depletion of Gal-3 in mouse tumor and human tumor cells reduced the levels of CD47 through inhibition c-Myc expression and its binding to the promoter of CD47. Additionally, tumor-derived Gal-3 were found critical for tumor progression accompanied with protected GAC cells from phagocytosis by macrophages. Gal-3 deficiency resulted in a marked reduction in M2 macrophages and elevated T cell effector activity. Dual blockade of Gal-3 and CD47 significantly decreased tumor growth by promoted phagocytosis, repolarization of macrophage and boosted T cell immune responses in the tumor microenvironment. Our data uncover that Gal-3 synergizes with CD47 to suppress phagocytosis and orchestrates immunosuppressed milieu in the tumor microenvironment warrants combination therapeutic strategy in GAC patients with peritoneal metastases.

Citation Format: Yibo Fan, Shumei Song, Yuan Li, Shilpa Dhar, Jiankang Jin, Xiaodao Yao, Ruiping Wang, Ailing Wang, Melissa Pool Pizzi, Jingjing Wu, Katsuhiro Yoshimura, Lang Ma, Samir Hanash, George Calin, Linghua Wang, Michael Curran, Jaffer Ajani. Galectin-3 synergizes with CD47 to suppress phagocytosis and T cell immunity in peritoneal metastases of gastric adenocarcinoma. [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2023; Part 1 (Regular and Invited Abstracts); 2023 Apr 14-19; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2023;83(7_Suppl):Abstract nr 4433.
Abstract 444: Inhibition of GPR68 signaling activated downstream of the Warburg effect induces cell death and enhances radiosensitivity in diverse cancer cell types
Authors: Neitzel Leif R. , Lee Han G. , Shukla Hem , Mahmood Javed , Williams Charles H. , Hong Charles C.
Published: April 2023

Introduction: The Warburg effect, a phenomenon in which cancer cells produce energy via aerobic glycolysis, is a hallmark of cancer. A key physiological consequence of the Warburg effect is lactate secretion, which acidifies the tumor milieu, thought to confer tumor resistance to chemotherapy and radiotherapy. Yet, the molecular mechanism of this resistance is not well understood. Ogremorphin, a small molecule inhibitor of an extracellular pH sensing receptor, GPR68, was found to inhibit the growth and survival of Glioblastomas. However, the consequences of GPR68 inhibition on carcinomas alone and in combination with radiation are unclear. The aim of this study was to investigate the effect of ogremorphin alone and in combination with radiation on multiple cancer cell lines.

Methods: 1) siRNA and CRISPR-interference knockdown of GPR68 in two carcinoma cell lines Panc02 (pancreatic ductal adenocarcinoma) and A549 (lung cell carcinoma). 2) Treatment of pancreatic and lung carcinomas with GPR68 inhibitor ogremorphin alone and with ionizing radiation. 3) Cell viability (MTT), clonogenic assays, and cell cycle analysis via flow cytometry were performed. 4) Lipid peroxidation study for induction of ferroptosis. The interaction between ogremorphin and radiation was evaluated using the combination index.

Results: Knockdown of GPR68 had significant effects on cell survival. Exposures of ogremorphin in clonogenic assays indicated a significant effect in reducing colony size, but not on the number of colonies, in both carcinoma cell lines. Moreover, ogremorphin significantly increased the efficacy of irradiation on the reduction of colony size. Although ogremorphin had no effect on cell cycle alone, it had a synergistic increase in G2 phase cells with radiation. Finally, GPR68 inhibition indicated synergistic activity with radiation for the induction of ferroptosis.

Conclusion. GPR68 inhibition induces ferroptosis in cancer cells on its own, but also synergistically with ionizing radiation. This study suggests that GPR68 is a key cancer survival pathway activated by the acidic tumor milieu/Warburg effect, and that the ogremorphin class of molecules may be suitable candidates for combination therapy in lung and pancreatic carcinoma.

Citation Format: Leif R. Neitzel, Han G. Lee, Hem Shukla, Javed Mahmood, Charles H. Williams, Charles C. Hong. Inhibition of GPR68 signaling activated downstream of the Warburg effect induces cell death and enhances radiosensitivity in diverse cancer cell types [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2023; Part 1 (Regular and Invited Abstracts); 2023 Apr 14-19; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2023;83(7_Suppl):Abstract nr 444.
Abstract 4449: The expression and immune regulation of thymic stromal lymphopoietin in non-small cell lung cancer development and progression
Authors: Chen Chung-Yu , Chen Ying-Yin , Peng Hung-Chueh , Ye Yi-Ling
Published: April 2023

Purpose: Previous studies have shown thymic stromal lymphopoietin (TSLP) to be an important driver of type 2 inflammation. A new and unexpected function for TSLP has been found in the induction and progression of a variety of tumors. This study conducted TSLP performance in lung cancer patients and the correlation of the clinical pathological data.

Materials and Methods: The patients with pathological diagnosis of lung cancer were enrolled. The percentage of CD19 + B cells was determined in the lymphocyte gate and counted via flow cytometry. Relevant serum cytokines including TSLP were analyzed using LEGENDplex software. In vitro study was conducted by A549 cell line and the mRNA expression change of TSLP was extracted by Reverse Transcriptase PCR (RT-PCR).

Results: When comparing the frequencies of B cell subsets between early-stage (N = 31) and late-stage lung cancer group (N = 29), the frequency of memory B cell (17.3±6.1% vs 23.4±15.1%, p=0.042), CD27+IgM+ B cell (1.8±1.3% vs 2.9±2.3%, p=0.030), class-switched B cell (15.5±5.5% vs 22.2±14.9%, p=0.023), class-switched memory B cell (69.9±11.3% vs 59.9±19.7%, p=0.017) and plasmablast (8.5±7.3% vs 18.0±20.8%, p=0.019) were significantly different between groups. The patients with higher frequency of class-switched B cell had significantly worse prognosis than low frequency (HR 3.054, 95% CI 1.007-9.262, p=0.049). The level of IL-1B, IL-6, IL-10, IL-12, IL-13, and TSLP were significantly higher in the late-stage cancer patients. Simulation in vitro test with human lung A549 cell line was performed, the expression of short-form TSLP (sfTSLP) level decreased with the higher anti-cancer drug as Pemetrexed concentration.

Conclusions: The increase proportion of CD27+CD38- (class-switched) B cell could be an independent, poor prognostic factor for NSCLC patients. TSLP signaling is addressed to the anti-cancer therapeutic response. Therefore, TSLP and relevant cytokines may be involved directly and indirectly in shaping the inflammatory status of the tumor microenvironment.

Citation Format: Chung-Yu Chen, Ying-Yin Chen, Hung-Chueh Peng, Yi-Ling Ye. The expression and immune regulation of thymic stromal lymphopoietin in non-small cell lung cancer development and progression. [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2023; Part 1 (Regular and Invited Abstracts); 2023 Apr 14-19; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2023;83(7_Suppl):Abstract nr 4449.
Abstract 4462: Immunoregulatory effects of NNMT-expressing cancer-associated fibroblasts
Authors: Heide Janna , Piffko Andras , Bilecz Agnes J. , Teich Ethan A. , Schweizer Lisa , Alhunayan Sayed R. , Yang Kaiting , Lengyel Ernst
Published: April 2023

Nicotinamide N-methyltransferase (NNMT) is highly expressed in the stroma of several malignancies and has been shown to drive the transformation of resting fibroblasts into cancer-associated fibroblasts (CAFs) through epigenetic changes. Given that CAFs can promote immunosuppression in cancer, the aim of our study was to investigate the effect of NNMT-expressing CAFs on the tumor-immune environment.

Using a co-culture system of normal fibroblasts and activated PBMCs, we found that IFN-γ produced by PBMCs upregulates NNMT expression in fibroblasts. We confirmed this mechanism in vivo in IFN-γ knockout mice and identified T cells as the main source of IFN-γ in the tumor. Gene expression and protein analysis showed that upregulation of NNMT in CAFs induces the secretion of the chemokine CXCL1, a critical chemoattractant for immunosuppressive myeloid-derived suppressor cells (MDSCs). Upregulation of NNMT induces CXCL1 secretion directly through promoter hypomethylation, and in vitro migration assays show that NNMT-expressing CAFs recruit high numbers of MDSCs. Using spectral flow cytometry, we analyzed the tumor-immune cell infiltration in whole-body NNMT-knockout mice and discovered that NNMT knockout of the stroma significantly reduced the abundance of MDSCs and increased the number of functional CD8+ T cells. Increased T cell cytotoxicity was associated with a significant reduction in tumor burden in the syngeneic ID8 ovarian and MC38 colon cancer models. These findings were confirmed after adoptive transfer of NNMT-wildtype immune cells into irradiated whole-body NNMT-knockout mice. Moreover, in scRNA-seq data of various human malignancies, we found a strong correlation between NNMT and CXCL1 expression in CAFs.

In summary, our results support a model in which cancer cells induce IFN-γ secretion by T-cells, thereby upregulating NNMT expression in fibroblasts, leading to CXCL1 secretion. CXCL1 secreted by CAFs recruits MDSCs to the tumor, which in turn suppress the anti-tumor T cell response.

Citation Format: Janna Heide, Andras Piffko, Agnes J. Bilecz, Ethan A. Teich, Lisa Schweizer, Sayed R. Alhunayan, Kaiting Yang, Ernst Lengyel. Immunoregulatory effects of NNMT-expressing cancer-associated fibroblasts. [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2023; Part 1 (Regular and Invited Abstracts); 2023 Apr 14-19; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2023;83(7_Suppl):Abstract nr 4462.
Abstract 4464: Comparison of two DTH models for T cell-mediated immunity in preclinical screen
Authors: Cai Yingying
Published: April 2023

Introduction: Delayed-type hypersensitivity (DTH) involves the recruitment of T cells into tissues and the activation by antigen-presenting cells to produce cytokines, which further activates local endothelial cells and recruits macrophages, resulting in tissue erythema, swelling and inflammation. DTH models in mice are classical in-vivo T cell-mediated immunity screening models for atopic dermatitis, asthma and autoimmune diseases. Different antigens and haptens have been used to establish DTH models with different Th1/Th2 profiles. More immunity characterization of the most widely used DTH models need to be revealed.

Results: We established DNFB, Oxazolone induced DTH models in both BALB/c and C57BL/6 mice with ear thickness as the readout. At the termination, we measured Th1 (ie, IFN-γ)/Th2 (ie, IL-4) profile, pro-inflammatory cytokines including IL-13, TNF, and IL-17 as well as myeloperoxidase (MPO) activity from mice ears as indices to score the degree of inflammation. Oxazolone in BALB/c exhibited more repeatable and robust window in Th1/Th2 balance and cytokines profiles. The comparison of two DTH models were further validated by the pathological analysis for cell infiltration in mouse ears and immune-phenotyping in spleen by FACS.

Conclusion: The comparison of DNFB and Oxazolone induced DTH models characterized the disease progression and established a more comprehensive preclinical DTH model with well-defined Th1/Th2 profile for the drug efficacy evaluation in the T cell-mediated immune response.

Key words: Delayed-type hypersensitivity, mice model, T cell-mediated

Citation Format: Yingying Cai. Comparison of two DTH models for T cell-mediated immunity in preclinical screen. [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2023; Part 1 (Regular and Invited Abstracts); 2023 Apr 14-19; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2023;83(7_Suppl):Abstract nr 4464.
Abstract 4465: Evaluation of tumor draining lymph nodes in dogs with spontaneously arising osteosarcoma using single-cell sequencing
Authors: Brill Samuel A. , Ammons Dylan T. , Avery Anne C. , Thamm Douglas H.
Published: April 2023

Tumor draining lymph nodes (TDLNs) function as an anatomic niche which primes immune cells to generate anti-cancer responses. While tumors are frequently staged based on the presence or absence of TDLN metastasis, mechanisms for how tumors impact the TDLN microenvironment and immunologic responses are incompletely understood. Dogs with spontaneously arising tumors provide an opportune model to study immuno-oncology, sharing many similarities to human patients with their environment, clinical care, and pathobiology. To evaluate how tumors impact TDLN responses, we conducted single cell RNA sequencing (scSeq) on cryopreserved TDLNs of dogs with osteosarcoma (n=4) and healthy controls (n=2). We obtained an average of 3,984 cells per sample (range 1,603-5,259) with a total of 4,780 cells from healthy samples and 19,121 cells from TDLN samples. All samples were integrated into one dataset, then unsupervised clustering was completed. Each unique cell cluster was assigned an identity using canonical markers and reference mapping to human datasets. Following cell classification, we observed notable changes in gene expression across all cells isolated from the LNs, with most differences arising from changes to the transcriptome of myeloid cells. Although still preliminary, the data also demonstrate differences in the abundance of T, B, and myeloid cells between healthy and tumor draining LNs. Taken together, this dataset provides indications of the changes TDLNs undergo in dogs with osteosarcoma.

Citation Format: Samuel A. Brill, Dylan T. Ammons, Anne C. Avery, Douglas H. Thamm. Evaluation of tumor draining lymph nodes in dogs with spontaneously arising osteosarcoma using single-cell sequencing. [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2023; Part 1 (Regular and Invited Abstracts); 2023 Apr 14-19; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2023;83(7_Suppl):Abstract nr 4465.
Abstract 4588: Vascular regulation of T-cell exclusion in melanoma
Authors: Park Ha-Ram , Shiva Anahita , Cummings Portia , Carvajal Richard , Thurston Gavin , An Joon-Yong , Lund Amanda W. , Yang Hee Won , Kim Minah
Published: April 2023

T-cell position in the tumor microenvironment determines the probability of target encounter and tumor killing. CD8+ T-cell exclusion from the tumor parenchyma is associated with poor response to immunotherapy and yet the biology that underpins this distinct pattern remains unclear. Here we show that the vascular destabilizing factor angiopoietin-2 (ANGPT2) causes compromised vascular integrity in the tumor periphery, leading to impaired T-cell infiltration within tumor. We analyzed the spatial regulation of ANGPT2 in whole tumor cross-sections and its impact on CD8+ T-cell position, vascular integrity, and response to immunotherapy using syngeneic murine melanomas. We found that T-cell exclusion was associated with peritumoral expression of ANGPT2 and elevated vascular leakage in human and murine melanomas. Both pharmacological and genetic blockade of ANGPT2 promoted CD8+ T-cell infiltration to the tumor core, exerting antitumor effects. Importantly, the reversal of T-cell exclusion following ANGPT2 blockade not only enhanced response to anti-PD-1 therapy in immunogenic, therapy responsive mouse melanomas, but also rendered non-responsive tumors susceptible. Therapeutic response after ANGPT2 blockade, driven by improved CD8+ T-cell infiltration, coincided with TIE2 signaling activation and increased vascular integrity in the tumor periphery, indicating that maintenance of vascular integrity within tumors is necessary for robust response to immunotherapy. These data highlight ANGPT2/TIE2 signaling as a key mediator of T-cell exclusion and a promising target to potentiate checkpoint inhibitor efficacy in melanoma.

Citation Format: Ha-Ram Park, Anahita Shiva, Portia Cummings, Richard Carvajal, Gavin Thurston, Joon-Yong An, Amanda W. Lund, Hee Won Yang, Minah Kim. Vascular regulation of T-cell exclusion in melanoma. [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2023; Part 1 (Regular and Invited Abstracts); 2023 Apr 14-19; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2023;83(7_Suppl):Abstract nr 4588.
Abstract 4593: Glioblastoma tumors that respond poorly to bevacizumab therapy show upregulation of an angiogenesis gene set
Authors: Lodha Roshan , Muller-Greven Gaelle , Guo Lydia , Gladson Candece
Published: April 2023

Glioblastoma (GBM) is the most common primary brain tumor in adults with a 15-month median survival, despite surgical-resection and radio-chemotherapy, and a recurrence rate of 90%. Despite improving survival in only a small percentage of patients, bevacizumab, a monoclonal antibody toward vascular endothelial growth factor-A, is frequently used to treat recurrent-GBM. To find predictors of poor-response to bevacizumab, we performed RNA-sequencing on multiple GBM patient-derived xenograft (PDX) tumors after orthotopic propagation in athymic nude mice. The study was repeated, and once PDX-tumors were established, mice were treated with bevacizumab and grouped based on survival. Bioinformatic-analysis of RNA-sequencing data demonstrated differential gene expression in tumors that were poor-responders to bevacizumab (no survival change) as compared to tumors that were good-responders (longer survival), along with upregulation of an angiogenesis gene set in poor-responders. Within this gene set, multiple genes known to be regulated by the early growth response 1 (EGR1) transcription factor were identified; CHRNA7 (cholinergic-receptor-nicotinic-α7-subunit) was selected for further study based on the reported role in promoting cancer cell migration and proliferation. Results were validated at the protein level using immunohistochemical staining. Additionally, Sry-box transcription factor-10 (SOX10) showed upregulation in poor-responders, potentially driven by impaired proteosome degradation. SOX10 is known to promote mural-cell coverage of neovasculature, cancer cell migration, and glioma development when cooperating with platelet-derived growth factor-B (PDGFB). In summary, GBM PDX-tumors with upregulated expression of an angiogenesis gene set and of two transcriptional regulators (EGR1 and SOX10) demonstrated a poor response to bevacizumab; upregulation of these genes could be used to predict bevacizumab response.

Citation Format: Roshan Lodha, Gaelle Muller-Greven, Lydia Guo, Candece Gladson. Glioblastoma tumors that respond poorly to bevacizumab therapy show upregulation of an angiogenesis gene set. [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2023; Part 1 (Regular and Invited Abstracts); 2023 Apr 14-19; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2023;83(7_Suppl):Abstract nr 4593.
Abstract 46: High-plex spatial transcriptomic characterization of canine tumor tissue
Authors: Church Sarah E. , London Cheryl , Toedebusch Christine M. , Sutton Ben , Weigel Sarah , Piazza Erin
Published: April 2023

Background: Combination therapy to treat hematological and solid malignancies including chemotherapy, radiation, targeted and immunotherapy all hold huge potential for eliciting clinical responses. Informative pre-clinical testing of these approaches can be greatly facilitated using immune competent animals with spontaneous tumors. Pet dogs are immunologically outbred, immune competent and develop spontaneous tumors such as non Hodgkin’s lymphoma, glioblastoma, osteosarcoma, urothelial carcinoma and melanoma that share remarkable clinical, biological and genetic features with their human counterparts. As such, pre clinical testing of therapeutic approaches in dogs with cancer promises to accurately inform human clinical trial design. For this comparative approach to provide maximum information to accelerate human clinical translation of novel combination therapies and identify correlative biomarkers of therapeutic response, it is necessary to develop research tools for deep interrogation of the canine tumor microenvironment (TME). Here we present spatial transcriptomic analysis of multiple canine tumor and tissue types using GeoMx® digital spatial profiler (DSP) Canine Cancer Atlas (CCA) panel.

Methods: FFPE slides or tissue microarrays were used to profile tumor and normal tissue from canines. Each slide was stained with tissue specific immunofluorescent antibodies, including Pan-cytokeratin, CD45, Vimentin, IBA1, CD3, and/or CD68. Regions of interest were selected to assess the TME and normal tissue as possible. Slides were then run on the DSP using the CCA panel that contains 1900-canine specific genes using standard DSP methods.

Results: We were able to spatially detect over 1700 genes across multiple tissue types from canines, including osteosarcoma, glioblastoma, melanoma and normal tissues. Genes were detected in spatial compartments including malignant tumor, tumor stroma and normal tissue.

Conclusions: Together the GeoMx CCA allow for interrogation of the TME of multiple tumor types and has the potential to inform spatial biomarkers for response to therapy, as well as translate the effectiveness of these therapies to humans.

Citation Format: Sarah E. Church, Cheryl London, Christine M. Toedebusch, Ben Sutton, Sarah Weigel, Erin Piazza. High-plex spatial transcriptomic characterization of canine tumor tissue [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2023; Part 1 (Regular and Invited Abstracts); 2023 Apr 14-19; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2023;83(7_Suppl):Abstract nr 46.
Abstract 4644: Modeling the effects of niche specific microenvironmental changes on patient-derived organoids
Authors: von Schlichting Jennifer , Peidli Stefan , Morkel Markus , Blüthgen Nils , Simon Leona
Published: April 2023

Here, we present an approach to study the effects of microenvironmental changes on inter- and intratumoral heterogeneity using colorectal cancer (CRC) patient-derived organoids. We identified relevant paracrine interactions between stromal and tumor cells based on the expression of ligand-receptor pairs in single cell-RNAseq data of 12 colorectal cancer patients. Physiological relevance was tested by adding CRC stroma-derived ligands to patient-derived organoids. A systematic screening of different conditions was enabled by multiplexed mass cytometry and scRNAseq analysis. We also aimed to model the effects of a changing extracellular matrix composition, by supplementing the laminin/collagen IV rich environment with other known ECM proteins such as collagen I to identify the impact of a changing substrate on cell plasticity. We found that ECM parameters as well as niche specific paracrine factors affect proliferation, differentiation, and developmental trajectories of colorectal cancer patient-derived organoids. We hypothesize that supplementing patient-derived organoids in vitro with physiologically relevant factors and substrates may expand the phenotypic space in which organoid cells differentiate, resulting in a more versatile preclinical model system.Our data provide insights into non-genetic sources of inter- and intratumoral heterogeneity resulting in differential drug responses. Additionally, they can serve as guidelines to improve existing patient-derived colorectal cancer models and provide a feasible approach to address common limitations in organoid culture.

Citation Format: Jennifer von Schlichting, Stefan Peidli, Markus Morkel, Nils Blüthgen, Leona Simon. Modeling the effects of niche specific microenvironmental changes on patient-derived organoids. [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2023; Part 1 (Regular and Invited Abstracts); 2023 Apr 14-19; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2023;83(7_Suppl):Abstract nr 4644.
Abstract 4647: Potential of TIMP3 regulating the versican status in the colorectal cancer tumor microenvironment
Authors: Field Anna , Kraus Sean , Matkowskyj Kristina , Deming Dustin , Pasch Cheri A. , Zhang Wei
Published: April 2023

Background: In colorectal cancer (CRC), versican (VCAN) accumulation has been found to negatively correlate with CD8+ tumor infiltrating lymphocyte (TIL) abundance and its proteolysis into versikine (Vkine) positively correlates with TIL abundance. TIMP3 has demonstrated the ability to inhibit several ADAMTS proteases able to cleave VCAN to Vkine. Here we investigate the correlation of TIMP3 accumulation in CRCs with the VCAN status and CD8+ T cell infiltration.

Methods: Matched primary and metastatic CRC samples from 156 patients who had resected oligometastatic CRC were stained for VCAN, Vkine, TIMP3, and CD8+ TILs. VCAN, Vkine, and TIMP3 were scored using an intensity binning system ranging from 0-3+ and T cells in the epithelium were reported per high power field (HPF). Tumors were designated as high (2 or 3+) or low (0 or 1) for both VCAN and Vkine and positive (+; 1-3) or negative (0) for TIMP3. Tumors designated VCAN low and Vkine high are considered VCAN proteolytic predominant (VPP) and all other combinations are considered VCAN proteolytic weak (VPW).

Results: Across the whole dataset, 52% of cancers were VCAN high and 24% were VPP. TIMP3 was detected in 33% of cancers. The frequency of TIMP3 was similar between primary and metastatic tumors (39% and 29%, respectively). Of the VCAN high cancers, 42% were TIMP3+ whereas 61% of the TIMP3+ samples are VCAN high. Of the Vkine high cancers, 44% were TIMP3+ and 36% of VPP cancers were TIMP3+. The prevalence of the VPP phenotype was similar between the TIMP3+ and negative cancers. Additionally, among the TIMP3+ samples 68% were Vkine high versus 46% in TIMP3 negative cancers (p=0.001). VCAN proteolysis correlates with higher CD8+ TIL abundance, thus we assessed the number of CD8+ TILs in TIMP3+ and negative tumors. The TIMP3+ cancers had a median of 2 CD8+ TILs/HPF and the TIMP3 negative cancers has a median of 3 CD8+ TILs/HPF. For VPP cancers, those that were also TIMP3+ had a median of 3 CD8+ TILs/HPF compared to TIMP3 negative cancers which had a median of 6 CD8+ TILs/HPF (p=0.2).

Conclusions: TIMP3 correlates with higher levels of VCAN accumulation but does not correlate with lower levels of proteolysis. Future studies are needed to better clarify the impact of TIMP3 and other mediators in regulating VCAN accumulation and proteolysis in the tumor microenvironment.

Citation Format: Anna Field, Sean Kraus, Kristina Matkowskyj, Dustin Deming, Cheri A. Pasch, Wei Zhang. Potential of TIMP3 regulating the versican status in the colorectal cancer tumor microenvironment. [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2023; Part 1 (Regular and Invited Abstracts); 2023 Apr 14-19; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2023;83(7_Suppl):Abstract nr 4647.
Abstract 466: A potential pro-oncogenic role of SERPINB4 in melanoma
Authors: Chhabra Gagan , Shirley Carl A. , Robarge Rachel K. , Rizvi Hassan A. , Ndiaye Mary A. , Ahmad Nihal
Published: April 2023

Despite recent therapeutic advancements, melanoma remains one of the deadliest forms of skin cancer. Melanoma metastasis and recurrence is associated with the existence of heterogeneous cell populations within tumors that have the ability to both initiate metastatic programs and bypass immune recognition. Tumor cells achieve immune evasion by interfering with immune signaling to limit immune cell infiltration, allowing tumor cell proliferation and growth. SERPINB4 (Squamous Cell Carcinoma Antigen 2 or SCCA2), a serine protease inhibitor, has been shown to regulate the host immune response against tumor cells by inhibiting immune cell activity. Interestingly, certain cancer cell types have been reported to overexpress SERPINB4. However, the expression profile and the functional significance of SERPINB4 in melanoma is not known. In this study, employing ‘The Cancer Genome Atlas’ (TCGA), we found that SERPINB4 is overexpressed and significantly associated with lower overall survival in patients with melanoma. Next, employing a panel of human melanoma cell lines, we demonstrated that SERPINB4 is overexpressed in melanoma cells when compared to normal human melanocytes. In addition, CRISPR knockout of SERPINB4 in high-SERPINB4-expressing A375 melanoma cells resulted in a significant decrease in proliferation of melanoma cells in comparison to control cells. Conversely, forced overexpression of SERPINB4 in low-SERPINB4-expressing SK-MEL-2 and SK-MEL-28 melanoma cells significantly increased melanoma cell proliferation. Moreover, RT-qPCR analyses suggested that SERPINB4 overexpression significantly increased, while its knockout significantly decreased epithelial-mesenchymal transition (EMT) markers (N-cadherin, β-catenin, Snail and Zeb1) in melanoma cells. Additionally, SERPINB4 manipulation significantly modulated the expression of genes involved in antigen-presentation (MHC Class I molecules, HLA-A, -B, and -C) in melanoma cells. Taken together, our data suggested a potential oncogenic role of SERPINB4 in melanoma. Our study also suggested that SERPINB4 might be a promising therapeutic target for melanoma. Indeed, additional in vitro mechanistic and in vivo studies are needed to firmly establish the mechanism and therapeutic potential of SERPINB4 for melanoma management.

Citation Format: Gagan Chhabra, Carl A. Shirley, Rachel K. Robarge, Hassan A. Rizvi, Mary A. Ndiaye, Nihal Ahmad. A potential pro-oncogenic role of SERPINB4 in melanoma [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2023; Part 1 (Regular and Invited Abstracts); 2023 Apr 14-19; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2023;83(7_Suppl):Abstract nr 466.
Abstract 4684: Genomic and transcriptomic analysis of a diffuse pleural mesothelioma patient-derived xenograft library
Authors: Sen Triparna
Published: April 2023

Background: Despite recent treatment advances, malignant pleural mesothelioma (MPM) is an aggressive, recalcitrant malignancy. Currently, the histologic subtype (epithelioid/non-epithelioid/biphasic) is the primary prognostic factor; other potential biomarkers to guide therapeutic strategies remain elusive. Even with multimodality therapies, recurrence is high in early-stage disease. In the unresectable/metastatic setting, there are only two FDA-approved regimens, both in the first-line setting: cisplatin/pemetrexed and ipilimumab/nivolumab. Unfortunately, most who respond to first-line treatment experience disease progression within a year. Therapeutic and diagnostic advances in DPM are hindered by a paucity of well-annotated preclinical models which can faithfully recapitulate the complex genomic interplay of the disease.

Methods: We established a library of patient-derived xenografts (PDX) from patients with DPM. We performed multi-omic analyses on available PDX and patient samples to deconvolute the mutational landscapes, global expression profiles, and molecular subtypes. Targeted next-generation sequencing (NGS; MSK-IMPACT), immunohistochemistry, and histologic subtyping were performed on all available samples. RNA-sequencing was performed on all available PDX samples. Clinical outcomes and treatment history were annotated for all patients. Platinum-doublet progression free survival (PFS) was determined from the start of chemotherapy until radiographic/clinical progression and grouped into < or ≥ 6 months.

Results: The mutational landscapes of PDX models strongly correlated with paired tumor samples. There were some differences in CDKN2A/B mutations and relative enrichment of NF2 with fewer BAP1 alterations, the significance of which is being investigated. When compared by histological subtype, we observed an upregulation of genes involved in NOTCH and EMT signaling in the epithelioid models. Models derived from patients with shorter overall survival or poor response to platinum doublet had higher expression of WNT/β-catenin signaling, hedgehog pathway, and epithelial-mesenchymal transition signaling as well as downregulation of immune-activation pathways, including type I and II interferon signaling and inflammatory response pathways.

Conclusions: This library of MPM PDXs, the largest to date, effectively mimics human disease and provides unprecedented insight into the genomic, transcriptomic, and protein landscape of MPM. These PDX models will inform future clinical investigations and provide an important new preclinical resource.

Citation Format: Triparna Sen. Genomic and transcriptomic analysis of a diffuse pleural mesothelioma patient-derived xenograft library. [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2023; Part 1 (Regular and Invited Abstracts); 2023 Apr 14-19; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2023;83(7_Suppl):Abstract nr 4684.
Abstract 4696: Characterizing CD39 and CD73 cell subtypes in the tumor microenvironment using MultiOmyx
Authors: Todorov Courtney , Gallagher Kevin , Wortman Juliana , Nunns Harry , Parnell Erinn , Leones Eric , Yeung Elaine , Russell Blair , Sahafi Flora , Au Qingyan
Published: April 2023

CD39 and CD73 are membrane bound enzymes that function together to convert extracellular adenosine 5’-triphospate (ATP)/adenosine diphosphate (ADP) into adenosine. CD39 first catalyzes ATP/ADP into adenosine monophosphate (AMP), which is then converted by CD73 into adenosine. Accumulation of extracellular adenosine creates an immunosuppressive tumor environment and facilitates tumor growth and metastasis. Conversely, the presence of ATP promotes a pro-inflammatory, tumor-suppressive environment. Suppression of CD73/CD39 activity and reduction of extracellular adenosine has been shown to support an antitumor immune response. Therefore, the targeting of CD73 and CD39, both individually as well as in combination with immune checkpoint inhibitors for biomarkers such as PD-1 and CTLA-4, is an emerging strategy for cancer therapeutics. CD73/CD39 have been described in multiple cell types, including tumor cells, fibroblasts, endothelial cells, tumor infiltrating lymphocytes (TILs), myeloid cells, and natural killer (NK) cells. While known to be expressed on multiple cell types, the spatial characterization of CD39 and CD73 in the tumor microenvironment (TME) is still poorly understood. Characterizing the diversity of active CD73 and CD39 populations in the tumor environment will help improve development of targeted therapies for cancer treatment. The multiplex immunofluorescence (mIF) platform MultiOmyx will be used to define the distinct populations of cells expressing CD73 and/or CD39 in the TME. MuliOmyx is a proprietary mIF platform for the visualization and characterization of up to 60 protein biomarkers in a single FFPE section. Herein we report the design and use of a novel panel of commercially-available antibodies broad enough to characterize CD39 and CD73 subpopulations in the TME of a variety of tumor indications including lung, prostate, and colon cancer. Using this panel in combination with proprietary deep-learning based image analysis, CD39 and CD73 positive cells can be characterized into different immune and TME subtypes. Understanding of the variety and phenotype of CD39 and CD73 expressing cells in the TME is crucial to define the populations being targeted by therapies for cancer treatment.

Citation Format: Courtney Todorov, Kevin Gallagher, Juliana Wortman, Harry Nunns, Erinn Parnell, Eric Leones, Elaine Yeung, Blair Russell, Flora Sahafi, Qingyan Au. Characterizing CD39 and CD73 cell subtypes in the tumor microenvironment using MultiOmyx. [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2023; Part 1 (Regular and Invited Abstracts); 2023 Apr 14-19; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2023;83(7_Suppl):Abstract nr 4696.
Abstract 4733: Effect of KDM6A mutation in bladder cancer
Authors: Le Ninh B. , Qiu Hong , Fink Emily E. , Sona Surbhi , Ting Angela H. , Lee Byron H.
Published: April 2023

As of 2020, bladder cancer is the tenth most common cancer in the world, with a heavier toll exerted on males. Its higher tumor recurrence and progression rates cause major treatment challenges. One of the most frequent mutations in bladder cancer is in lysine-specific demethylase 6A (KDM6A). This gene encodes for ubiquitously transcribed tetratricopeptide repeat on chromosome X (UTX), which catalyzes the demethylation of di- and tri- methylated histones H3 lysine 27 and works as part of “COMPASS-LIKE” protein complexes to coordinate many transcriptional processes. The molecular basis for how KDM6A mutations promote carcinogenesis is still under investigation. In this study, we use single-cell RNA sequencing data from bladder cancers with and without KDM6A mutations in both human and mice to examine the effects of KDM6A mutations on the cellular composition and gene expression of tumor and normal bladders. In mice without tumors, KDM6A mutations seem to correlate with enrichment of macrophages in the bladder tissue microenvironment. The opposite is true, however, in the presence of tumors, as there were not only more macrophages but also more dendritic cells in wildtype mice. This pattern was also observed in human, suggesting differences in immune infiltrates in the tumor microenvironment are part of the phenotype of KDM6A mutation. Lastly, KDM6A mutations may also impact urothelial cell states at baseline, as KDM6A KO mouse bladders contain higher proportions of intermediate cell populations compared with their wildtype counterparts. Together, these observations contribute to a better understanding of the role KDM6A plays in bladder cancer.

Citation Format: Ninh B. Le, Hong Qiu, Emily E. Fink, Surbhi Sona, Angela H. Ting, Byron H. Lee. Effect of KDM6A mutation in bladder cancer. [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2023; Part 1 (Regular and Invited Abstracts); 2023 Apr 14-19; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2023;83(7_Suppl):Abstract nr 4733.