bluuebunny/crossref_metadata_2025 huggingface dataset:
Abstract 4246: ABL103, A novel T-cell engaging bispecific antibody, exhibits potent in vitro and vivo antitumor activity and low toxicity via B7-H4 dependent 4-1BB activation in tumor microenvironment
Authors: Park Kyungjin , Lee Yangsoon , Lim Saeyi , Park Kyeongsu , Kim Eunjung , Lee Hanbyul , Yoo Jiseon , Pak Youngdon , Kim Yeunju , Ko Minji , Won Jonghwa
Published: June 2022

B7-H4 (B7x, VTCN1), a member of the B7-family, is overexpressed in majority of cancer patients with ovarian, endometrial and breast cancers. B7-H4 expression was also observed in tumor associated macrophages and implicated as an immune checkpoint regulating T cell responses. Although B7-H4 expression in normal tissues is quite limited, tumor-specific immune response by ABL103 would minimize potential adverse effects. ABL103, B7-H4 Grabody-T, is a First-in Class bispecific antibody targeting B7-H4 and 4-1BB and augments T cell function by a dual mechanism, i.e., 1) blocks the B7-H4 mediated T cell inhibition, and 2) elicits superior T cell activation through B7-H4-dependent 4-1BB clustering. Our data showed that simultaneous binding of B7-H4 and 4-1BB by ABL103 led to potent in vitro T cell activation only in the presence of B7-H4 expressing tumor cells. In the established tumor model, ABL103 potently inhibited tumor progression in a dose-dependent manner and showed higher rate of complete remission (CR) at 2 and 10 mg/kg dose groups. Moreover, mice were protected from the tumor re-challenge 3 months after cessation of ABL103 treatment, suggesting long-term memory has been established. In 4-week pilot tox study using cynomolgus monkeys, ABL103 was tolerable up to 100 mg/kg dosed weekly with no ABL103-related toxicity observations. To understand the relationship of target expression, expression of B7-H4, CD4, CD8, PD-L1, and 4-1BB was examined in 142 ovarian cancer patient biopsies. About 83% of ovarian cancer patient tissues showed B7-H4 positive staining and 62% of them showed strong positive. In addition, the staining of 4-1BB as well as CD4 and CD8 T cells were observed in both tumor nest and stroma, suggesting that cross-linking of B7-H4 and 4-1BB is feasible in tumor microenvironment. Overall, ABL103 has a strong in vitro and vivo anti-tumor activity and good safety profile via B7-H4-dependent 4-1BB activation. This strongly suggests ABL103 is a promising therapeutic agent potentially benefitting patients with B7-H4 overexpression.

Citation Format: Kyungjin Park, Yangsoon Lee, Saeyi Lim, Kyeongsu Park, Eunjung Kim, Hanbyul Lee, Jiseon Yoo, Youngdon Pak, Yeunju Kim, Minji Ko, Jonghwa Won. ABL103, A novel T-cell engaging bispecific antibody, exhibits potent in vitro and vivo antitumor activity and low toxicity via B7-H4 dependent 4-1BB activation in tumor microenvironment [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 4246.
Abstract 4249: Quantitative PBMC ADCC bioassay and ADCC reporter bioassays for immunotherapy and SARS-CoV-2 monoclonal antibody development
Authors: Stecha Pete , Wang Jun , Garvin Denise , Gilden Julia , Grailer Jamison , Hartnett Jim , Fan Frank , Cong Mei , Cheng Zhi-jie Jey
Published: June 2022

Fc effector function is one of the main mechanisms of action (MoA) for therapeutic monoclonal antibodies (mAbs). Quantitative measurement of antibody-dependent cellular cytotoxicity (ADCC) is critically required for understanding the Fc function in mAb drug development. Despite the increasing interest and clinical success of the mAb therapeutic, it has been highly challenging to measure their ADCC activity in a reproducible and quantitative manner due to the lack of consistency in current methods that are based on primary PBMCs or NK cells and use tedious assay procedures. To improve ADCC assay precision so they can be validated as potency assay in cGMP laboratories, we developed reporter based ADCC bioassays using engineered effector cell line stably expressing a luciferase reporter and FcγRIIIa (V or F variant) to replace primary PBMC to overcome the assay variation. The ADCC reporter bioassays have been validated according to ICH guidelines by many laboratories and are demonstrated to be suitable for product release and stability studies in a quality-controlled environment. For early research and antibody characterization, we developed an improved PBMC ADCC assay using ADCC-prequalified PBMCs and engineered HiBiT target cells so they can measure the target specific lysis in ADCC. The PBMCs used in the study are isolated from prescreened blood donors and QC tested in ADCC assay. When HiBiT target cells are incubated with an antibody and PBMCs, HiBiT are released to the culture medium where it binds to LgBiT in the detection reagent to form a functional NanoBiT luciferase to generate luminescence signal. This new PBMC ADCC bioassay is simple, homogenous, highly sensitive, and gives a robust assay window. We demonstrate that it can quantitatively measure the potency for mAb drugs in cancer immunotherapy (e.g., rituximab, trastuzumab), and for anti-SARS-CoV-2 spike antibodies in antiviral drug development. Additionally, it shows antibody potency comparable with the ADCC reporter bioassay. In summary, the new PBMC ADCC bioassay using HiBiT target cells can be a valuable tool for early antibody discovery and characterization and also for method bridging study with ADCC reporter bioassay.

Citation Format: Pete Stecha, Jun Wang, Denise Garvin, Julia Gilden, Jamison Grailer, Jim Hartnett, Frank Fan, Mei Cong, Zhi-jie Jey Cheng. Quantitative PBMC ADCC bioassay and ADCC reporter bioassays for immunotherapy and SARS-CoV-2 monoclonal antibody development [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 4249.
Abstract 426: Development of oridonin-based proteolysis-targeting chimera (PROTAC) degraders as effective breast cancer therapeutics
Authors: Ma Ruixia , Wang Pingyuan , Xu Junhai , Xu Jimin , Fricke Doerte R. , Xue Yu , Kim Hyejin , Chen Haiying , Liu Xi , Zhou Jia , Shen Qiang
Published: June 2022

Natural products provide a wealth of resources as the molecular starting point for the development of therapeutic agents, including a plethora of anticancer drugs currently in clinical use. Oridonin is a natural diterpene compound enriched in the medicinal herb Rabdosia rubescens, and its anticancer property has been extensively studied. However, the potency of oridonin against triple-negative breast cancer (TNBC) limits its potential clinical translation. Previously, we developed a class of oridonin analogues including CYD0618 and others as effective anticancer agents to inhibit tumor growth of TNBC in vitro and in vivo. Intriguingly, CYD0618 has been shown to directly bind to and inhibit STAT3 transcription factor, which is constitutively activated in the majority of human cancers. Currently there are no FDA-approved direct STAT3 inhibitors in clinical use. Based on the proteolysis-targeting chimera (PROTAC) technology, our team designed novel oridonin- and CYD0618-based PROTACs in an attempt to achieve specific STAT3 binding and protein degradation. We determined anticancer effects of oridonin- and CYD0618-PROTACs on proliferation, colony formation, and apoptosis of the TNBC cell line MDA-MB-231. We found that oridonin- and CYD0618-based PROTACs inhibited the proliferation of MDA-MB-231 cells significantly, similarly effective as the lead compound CYD0618 and much more potent than oridonin. We then examined the expression of STAT3, pSTAT3 and protein biomarkers for tumor metastasis using Western blot. STAT3 and pSTAT3 protein levels were downregulated dose- and time-dependently in MDA-MB-231 cells treated with CYD0618 and CYD0618-PROTACs, respectively. Our results demonstrate impressive effects of CYD0618-based PROTACs against TNBC cells, primarily through direct targeting of STAT3. We are in the process of further characterizing the molecular profile of CYD0618-based PROTACs by studying its interaction with STAT3 and protein degradation as well as their efficacy against TNBC tumor xenografts in vivo.

Citation Format: Ruixia Ma, Pingyuan Wang, Junhai Xu, Jimin Xu, Doerte R. Fricke, Yu Xue, Hyejin Kim, Haiying Chen, Xi Liu, Jia Zhou, Qiang Shen. Development of oridonin-based proteolysis-targeting chimera (PROTAC) degraders as effective breast cancer therapeutics [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 426.
Abstract 483: Automated Ki67-LI assessment in prostate cancer using artificial intelligence in multiplex fluorescence immunohistochemistry
Authors: Blessin Niclas C. , Mandelkow Tim , Bady Elena , Simon Ronald , Hube-Magg Claudia , Lennartz Maximilian , Sauter Guido , Graefen Markus , Steurer Stefan
Published: June 2022

The Ki67 labeling index (Ki67-LI) is a strong prognostic marker in prostate cancer. Its analysis requires cumbersome manual quantification of Ki67 immunostaining in at least 200 tumor cells. To enable automated Ki67-LI assessment in routine clinical practice, we have developed and validated a framework for automated Ki67-LI quantification, which comprises three different artificial intelligence analysis steps and an algorithm for cell-distance analysis of multiplex fluorescence immunohistochemistry staining. The prognostic impact of the Ki67-LI was tested on a tissue microarray (TMA) containing one sample each of 12475 prostate cancers. A “heterogeneity TMA” containing 3 to 6 samples from different tumor areas was used to model Ki67 analysis of multiple different biopsies. The Ki67-LI provided strong and independent prognostic information in 11845 successfully analyzed prostate cancers (p<0.001 each). The analysis of the heterogeneity TMA revealed that the Ki67-LI of the sample with the highest Gleason score (AUC:0.68) was similarly prognostic as the mean Ki67-LI of all six foci (AUC:0.71 [p=0.24]). The combined analysis of the Ki67-LI and Gleason grades obtained on identical tissue spots showed that the Ki67-LI added significant additional prognostic information in case of classical ISUP grades (AUC:0.82 [p=0.002]) and quantitative Gleason grades (AUC:0.83 [p=0.018]). The Ki67-LI is a powerful prognostic parameter in prostate cancer, which can be efficiently analyzed in multiplex fluorescence IHC. In case of multiple cancer positive biopsies, the sole analysis of the worst biopsy can be sufficient.

Citation Format: Niclas C. Blessin, Tim Mandelkow, Elena Bady, Ronald Simon, Claudia Hube-Magg, Maximilian Lennartz, Guido Sauter, Markus Graefen, Stefan Steurer. Automated Ki67-LI assessment in prostate cancer using artificial intelligence in multiplex fluorescence immunohistochemistry [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 483.
Abstract 5006: Computational analysis of hypoxia-inducible factor 1-alpha and UBS109 in hepatocellular carcinoma
Authors: Merchant Neha Bahadur , Behera Santosh Kumar , Alam Afroz , Nagaraju Ganji Purnachandra , Basha Riyaz
Published: June 2022

Hypoxia-inducible factor 1-alpha (HIF-1α) is a heterodimeric transcription factor, which is elevated during hypoxic conditions in several malignancies. HIF-1α is known to stimulate hypoxia-responsive genes that are linked with malignant properties viz.: progression, metastasis, and resistance to therapy. The objective of the present study is to target HIF-1α by potential phytodrug, curcumin and its monocarbonyl analog UBS109 in hepatocellular carcinoma (HCC). Curcumin has been known to exhibit therapeutic properties for the management of HCC due to its powerful anti-inflammatory and antioxidant properties as well as its ability to regulate multiple signaling pathways. UBS109 possesses comparable antitumor and anti-inflammatory properties as curcumin without any of its limitations. To analyze the binding capacity and ligand efficacy of HIF-1α against curcumin and UBS109, computational approaches were applied including molecular docking and dynamics simulation that can provide detailed understanding of its implication in HCC treatment and a deeper analysis of the protein-drug interaction.

Citation Format: Neha Bahadur Merchant, Santosh Kumar Behera, Afroz Alam, Ganji Purnachandra Nagaraju, Riyaz Basha. Computational analysis of hypoxia-inducible factor 1-alpha and UBS109 in hepatocellular carcinoma [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 5006.
Abstract 5010: Comprehensive characterization of 11 prognostic alternative splicing events in ovarian cancer interacted with the immune microenvironment
Authors: Zheng Guixi
Published: June 2022

Alternative splicing (AS) events play a crucial role in the tumorigenesis and progression of cancer. Transcriptome data and Percent Spliced In (PSI) values of ovarian cancer patients were downloaded from TCGA database and TCGA SpliceSeq. Totally we identified 1,472 AS events that were associated with survival of ovarian serous cystadenocarcinoma (OV) and exon skipping (ES) was the most important type. Univariate and multivariate Cox regression analysis were performed to identify survival-associated AS events and developed the prognostic model based on 11-AS events. The immune cells and different response to cytotoxic T lymphocyte associated antigen 4 (CTLA-4) and programmed cell death protein 1 (PD-1) blockers in low-risk and high-risk group of OV patients were analyzed. Ten kinds of immune cells were found up-regulated in low-risk group. Activated B cell, natural killer T cell, natural killer cell and regulatory T cell were associated with survival of OV. The patients in low-risk group had good response to CTLA-4 and PD-1 blockers treatment. Moreover, a regulatory network was established according to the correlation between AS events and splicing factors (SFs). The present study provided valuable insights into the underlying mechanisms of OV. AS events that were correlated with the immune system might be potential therapeutic targets.

Citation Format: Guixi Zheng. Comprehensive characterization of 11 prognostic alternative splicing events in ovarian cancer interacted with the immune microenvironment [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 5010.
Abstract 502: The association of metformin dose and cancer survival in older adults with advanced cancer: SEER-Medicare analysis
Authors: Scarton Lisa , Jo Ara , Xie Zhigang , O’Neal LaToya J. , Pena Juan M. , George Thomas J. , Bian Jiang
Published: June 2022

Cancer and type 2 diabetes (T2DM) are common diseases that have serious health consequences. T2DM is known to increase the risk for the development of many of the most common cancers. Metformin, one of the most commonly used drugs to treat T2DM, is increasingly being used for its anticancer effects; however, the literature is limited on the effect of metformin dose on OS in patients with advanced cancer. We examined the relationship between metformin dose and OS in persons with both T2DM and stage IV lung, breast, colorectal, prostate, or pancreas cancers.

Methods: We used a retrospective study design with cox proportional hazards regression analysis of the 2007-2016 of the Surveillance Epidemiology and End Results-Medicare dataset.

Results: A total of 7,725 patients with T2DM and a least one targeted cancer was included in the analysis. Of those patients, 38.5% had been prescribed metformin. Patients who used metformin had significantly better OS in both unadjusted (Unadjusted HR, 0.73; 95% CI, 0.69-0.76; p < 0.001) and adjusted models (adjusted HR, 0.77; 95% CI, 0.73-0.81; p < 0.001). The OS between patients who took metformin with average daily dose ≥ 1000mg or < 1000mg were not statistically significant (aHR, 1.00; 95% CI, 0.93-1.08; p=0.90).

Conclusions: Metformin use, regardless of dose, increases OS in older adults with advanced cancer. Additionally, OS was not significantly different across race or ethnicity.

Citation Format: Lisa Scarton, Ara Jo, Zhigang Xie, LaToya J. O’Neal, Juan M. Pena, Thomas J. George, Jiang Bian. The association of metformin dose and cancer survival in older adults with advanced cancer: SEER-Medicare analysis [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 502.
Abstract 5030: Intra-patient tumor evolution analysis in dogs with many mammary tumors identifies signatures of tumor aggression in early-stage breast cancers
Authors: Graim Kiley
Published: June 2022

Despite tremendous improvements in breast cancer detection, it remains unclear which early-stage tumors will later become aggressive. Dogs provide a unique opportunity to unravel early-stage tumor evolution, as dogs frequently develop multiple naturally occurring mammary tumors. To identify which early-stage human breast tumors will later become malignant, we analyzed mutational profiles of mammary tumors from two canine cohorts to identify patterns of tumor evolution. We compared our dog tumor signatures with human breast cancers. We found that dogs and humans share many known cancer driver mutations. Additionally, three of the five dogs with at least five mammary tumors had a single driver mutation present in most of their tumors. Despite sharing patient-level exposures and environment, passenger mutations significantly differ between these tumors, suggesting independent primaries initiated by a catalyzing factor causing the same driver mutation in each tumor. However, tumors from the same dog with the same driver gene mutations rarely exhibit the same expression-based breast cancer subtypes. Our analysis highlights the mutational similarities of dog and human breast cancers and provides insight into their development.

Citation Format: Kiley Graim. Intra-patient tumor evolution analysis in dogs with many mammary tumors identifies signatures of tumor aggression in early-stage breast cancers [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2022; 2022 Apr 8-13. Philadelphia (PA): AACR; Cancer Res 2022;82(12_Suppl):Abstract nr 5030.
Abstract 5076: Evaluation of the CNS penetration of a next generation PARP inhibitor, AZD9574, in cynomolgus monkey using positron emission tomography
Authors: Pike Andy , Balazs Amber , Cselényi Zsolt , Degorce Sébastien L. , Ghosh Avipsa , Hande Sudhir M. , Johannes Jeffrey , Johnström Peter , Packer Martin J. , Schou Magnus , Zheng XiaoLan
Published: June 2022

The current clinically approved PARP inhibitors have limited subtype selectivity and are to some degree restricted in their ability to penetrate the central nervous system (CNS) due to efflux transporters, potentially limiting their efficacy in treating metastatic disease or primary tumors in the brain. The current study evaluated the potential of AZD9574, a next generation, PARP1 selective inhibitor/trapper, to penetrate the CNS in cynomolgus monkey, and its occupancy of the PARP1 enzyme, using positron emission tomography (PET). In vitro bidirectional efflux assay data suggested AZD9574 showed minimal substrate potential compared to the clinically approved PARP inhibitors. This was reflected in an increased ratio of unbound brain to unbound plasma concentration (Kpuu) in the rat of ~0.31. Therefore AZD9574 was taken forward into cynomolgus monkey PET studies. Firstly, the Kpuu was determined following dosing of [11C]AZD9574, co-administered with unlabeled drug to minimize the impact of specific binding. The high specific signal observed lead to the development of [11C]AZ3391, a PARP1 selective, CNS penetrant PET tracer, which was subsequently used to directly assess the PARP1 target engagement of AZD9574 in the brain. AZD9574 was found to show a Kpuu in cynomolgus monkeys of 0.79, close to unity with unbound plasma concentrations suggesting minimal CNS restriction. Furthermore, an i.v. infusion dose response study with AZD9574, conducted to examine its ability to block target occupancy by the PET tracer [11C]AZ3391, demonstrated a reduction in [11C]AZ3391 accumulation in whole brain. The resulting calculated occupancy of AZD9574 ranged from 17% for the lowest dose (0.003 mg/kg) to 95% for the highest dose tested (0.05 mg/kg). Comparable reduction in occupancy was seen for peripheral tissue, such as bone marrow, supporting the conclusion that AZD9574 shows minimal CNS restriction. These data show that AZD9574 is the first PARP inhibitor to reach the clinic which combines PARP1 selectivity, trapping and high CNS penetration in a single molecule and supports its development as a potential therapy for the treatment of metastatic disease and primary brain tumors.

Citation Format: Andy Pike, Amber Balazs, Zsolt Cselényi, Sébastien L. Degorce, Avipsa Ghosh, Sudhir M. Hande, Jeffrey Johannes, Peter Johnström, Martin J. Packer, Magnus Schou, XiaoLan Zheng. Evaluation of the CNS penetration of a next generation PARP inhibitor, AZD9574, in cynomolgus monkey using positron emission tomography [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 5076.
Abstract 5086: Effects of immune negative genes on immunotherapy combined anti-vascular for liver cancer
Authors: Zhu Xinhua , Chen Yanan , Zhang Junling , Huang Mengli , Xia Xihua
Published: June 2022

Background: In recent years, the application of immune checkpoint inhibitors (ICIs) therapies targeting the programmed cell death-1 (PD-1)/programmed cell death ligand 1 (PD-L1) or cytotoxic T lymphocyte (CTLA-4) pathway represents a major breakthrough in many types of cancer. However, low response rates has limited the clinical application of ICIs monotherapy for liver cancer.Anti-vascular combined ICIs are the preferred regiments in first-line systemic treatment of hepatocellular carcinoma (HCC) by NCCN. Therefore, strategies to improve efficacy through patient stratification, biomarker-directed therapies or combinations remain urgently needed. The purpose of this study was to explore potential genomic markers associated with “low response rates” in liver cancer with ICIs combination therapy.

Methods: Between July 2019 and April 2021, a total of 47 patients with advanced liver cancer treated with ICIs combined anti-vascular or anti-vascular alone therapy were admitted to Drum Tower Hospital. We retrospectively identified genomic markers, abstracted clinicopathologic features and treatment outcomes.

Results: 47 patients with advanced liver cancer treated with immunotherapy combined anti-vascular or anti-vascular were included in this analysis. 40 of 47patients (85.1%) were male and the median age was 58 years (range, 32 to 80 years). Most patients (85.1%) were HCC and 12 patients had at least one immune negative gene mutation (including CCND1, FGF19, FGF3, FGF4, KEAP1, JAK1, MDM4, DNMT3A, EGFR and STK11). Median follow-up was 206 days in immunotherapy combined anti-vascular group and 215 days in anti-vascular group. In the immunotherapy combined anti-vascular group, compared with wild-type, patients with immune negative gene mutations were significantly associated with poorer overall survival (P=0.021). In anti-vascular alone group, there was no significant difference in median OS for patients with/without immune negative gene (P=0.94). In addition, patients with immune negative gene mutations, OS of anti-vascular alone was better than immunotherapy combined anti-vascular (P=0.085).

Conclusion: Our study revealed that liver cancer with immune negative gene mutations have a worse prognosis than wild-type patients on immunotherapy combined anti-vascular. In addition, in patients with immune negative gene mutations, immunotherapy combined anti-vascular did not significantly improve prognosis compared to anti-vascular alone.

Citation Format: Xinhua Zhu, Yanan Chen, Junling Zhang, Mengli Huang, Xihua Xia. Effects of immune negative genes on immunotherapy combined anti-vascular for liver 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 5086.
Abstract 5087: KDR mutation as novel biomarker for immune checkpoint inhibitors in pan-cancer
Authors: Guo Renhua , Cui Yanan , Zhang Pengpeng , Liang Xiao , Xu Jiali , Liu Xinyin , Wu Yuemin , Wang Wei , Zhang Fang , Zhang Junling , Huang Mengli
Published: June 2022

Purpose: Immune Checkpoint Inhibitors (ICIs) have marked important milestones in cancer treatment, while few patients experience clinical benefits from them. Therefore, reliable predictive markers for ICIs are urgently needed. Nowadays, the combination of antiangiogenic therapy and ICIs exhibits encouraging efficacy. As a key protein of VEGFR signaling, KDR may also function to regulate immune responses beyond its role in angiogenesis.

Experimental Design: Pan-cancer immunotherapeutic patients from public cohorts including genomic data and clinical data were analyzed. Further analysis was performed on an internal validation data set including 67 non-small cell lung cancer (NSCLC). Investigation on underlying mechanism was performed in The Cancer Genome Atlas (TCGA) data via bioinformatics analysis.

Results: We found better responses to ICIs in patients who harbored KDR mutation from pan-cancer public datasets (discovery cohort: n=662; validation cohort: n=1333). Our NSCLC cohort verified the value of KDR mutation in predicting better ICI outcomes, including objective response rate (70.0% vs 22.81%, P=0.0057) and progression-free survival (HR=0.158, 95% CI, 0.045-0.773; P=0.007). KDR mutation was associated with increased TMB level, neoantigen burden, immune cellular activities, and decreased VEGFR signaling. Meanwhile, KDR mutation was indicative of an immune-hot status, characterized by higher expression of PD-L1 and abundance of cytotoxic lymphocytes.

Conclusions: Our work demonstrates that KDR mutation has the potential to become a predictive biomarker for ICIs.

Citation Format: Renhua Guo, Yanan Cui, Pengpeng Zhang, Xiao Liang, Jiali Xu, Xinyin Liu, Yuemin Wu, Wei Wang, Fang Zhang, Junling Zhang, Mengli Huang. KDR mutation as novel biomarker for immune checkpoint inhibitors in pan-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 5087.
Abstract 5096: Characteristics of tumor immune microenvironment in patients carrying comutations of DNA damage response pathways in esophageal cancer
Authors: Chen Hao , Xia Xihua , Zhang Junling , Chen Yanan , Huang Mengli
Published: June 2022

Background: Immunotherapy exhibits promising clinical efficacy in treating unresectable esophageal cancer, and several studies have demonstrated that both comutations in DNA damage response (DDR) pathways and tumor immune microenvironment(TIME) could serve as potential biomarkers for immune checkpoint blockade (ICB) treatment. However, the associtation between DDR comutations and TIME remains unclear.

Methods: Between April 2020 and September 2020, NGS and TIME analysis of tumor tissue from 22 patients with esophageal cancer were performed by 3D Medicines, Inc., a College of American Pathologists (CAP)-accredited and Clinical Laboratory Improvement Amendments (CLIA)-certified laboratory. We retrospectively investigated the association between genomic alterations and immune microenvironment features. Among eight DDR pathways, comutations of check point factors(CPF) plus base excision repair(BER), homologous recombination repair(HRR), mismatch repair(MMR) or Fanconi anemia(FA) were defined as "DDR co-mut+", while mutation of CPF or comutations of CPF plus nucleotide excision repair (NER) were defined as “DDR co-mut-”. In addition, the relationship between comutations of DDR pathways and tumor mutational burden (TMB) was also analyzed.

Results: 22 patients with esophageal cancer were included for analysis. 16 of 22 patients (72.7%) were male and the median age was 62 years old(range, 52 to 73 years). Based on the definition of DDR co-mut+ and DDR co-mut-, 13 patients were devided into DDR co-mut+ group, and 9 patients belonged to DDR co-mut- group. Gene alterations and TIME analysis revealed that patients with DDR co-mut+ showed a higher abundance of CD56dim natural killer cells in both tumor(p=0.0161) and stroma(p=0.028) compared with DDR co-mut- patients. No differences were observed in the densities of CD8+ T cells, M1-TAMs, and M2-TAMs between DDR co-mut+ and DDR co-mut- patients. TMB analysis displayed slightly higher in DDR co-mut+ group with no significant difference.

Conclusions: Patients with DDR co-mut+ showed better infiltration of CD56dim NK cells in tumor and stroma in esophageal cancer. Combination of DDR comutations with TIME features may be a more promising biomarker to guide immunotherapy.

Keywords: esophageal cancer; DNA damage response pathway; tumor immune microenvironment; tumor mutational burden

Citation Format: Hao Chen, Xihua Xia, Junling Zhang, Yanan Chen, Mengli Huang. Characteristics of tumor immune microenvironment in patients carrying comutations of DNA damage response pathways in esophageal 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 5096.
Abstract 5115: Combine high risk factors with circulating tumor DNA (ctDNA) to assist adjuvant chemotherapy (ACT) decision in Stage II colorectal cancer (CRC)
Authors: Yuan Xie Gui , Miao Liu , Li Huang Meng
Published: June 2022

Background: Approximately 10% to 15% have residual disease after surgery in patients with stage II CRC. Identification of this high-risk population and treatment with ACT could potentially reduce their risk of recurrence. However, the relative contribution of the factors used to define high risk stage II disease needs better understanding, although post-surgical ctDNA has been studied as a marker for an elevated risk of recurrence in stage I-III colorectal cancer.

Methods: Individual data from 3 early colorectal cancer clinical studies(Tie, 2016;Reinert, 2019; Chen, 2021) that included information on high-risk factors and post-surgical ctDNA status were collected and reanalysed for 595 patients with or without ACT.

Results: In stage II CRC patients with ctDNA-negative not treated with ACT form the Tie cohort, with high-risk status (n=42) showed a significantly reduced recurrence-free survival (RFS) compared to those with low-risk status(n=122) [HR, 3.283; 95%CI, 1.398 to 14.18; P = 0.0116], but in the ctDNA-negative patients with high-risk status, ACT (n=32) or not did not affect the RFS(P=0.1758). It meant false negatives of ctDNA made for bias of survival analysis results. Interestingly, in other two cohorts of stage II CRC patients with ctDNA-negative not treated with ACT, high-risk status showed no difference in RFS compared to low-risk status (p=0.3429 and 0.3173, respectively). In addition to the low sample size, the contribution of defined high-risk factors in prognosis was worth exploring which meant pseudo high-risk status also affect survival analysis results. And there were big differences in the high-risk factors included in the three cohorts especially in ctDNA-negative patients, such as the proportion of lymph nodes (LN) yield <12 seemed higher in Tie cohort.

Conclutions: It was not enough to predict recurrence depend on ctDNA status alone for stage II CRC. Different risk factors might have different prognostic weights so that different combinations of post-surgical ctDNA status and risk factors might be used to make more accurate ACT decision.

Citation Format: Xie Gui Yuan, Liu Miao, Huang Meng Li. Combine high risk factors with circulating tumor DNA (ctDNA) to assist adjuvant chemotherapy (ACT) decision in Stage II colorectal cancer (CRC) [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 5115.
Abstract 5119: Genomic landscape analysis in tumor tissue and circulating tumor DNA of Chinese patients with hepatocellular carcinoma
Authors: Yang Dinghua , Yu Sheng , Guo Baitang , Chen Tingting
Published: June 2022

Background: Hepatocellular carcinoma (HCC) is a leading cause of tumor-associated death worldwide, owing to its high 5-year postoperative recurrence rate and heterogeneity. Although Molecular profiling has been used to predict prognosis and guide targeted therapy, obtaining liver tissue biopsies was still a clinical challenge of advanced HCC patients. Circulating tumor DNA (ctDNA) testing as a novel and convenient detecting techniques, which can revealed comprehensive information regarding to tumor genome profiles. The purpose of the study was to analyze and compare the genomic landscape characteristics of tumor tissue and ctDNA in Chinese HCC.

Methods: 187 tumor tissue samples and 293 plasma samples from patients with HCC were collected from Nanfang hospital and 3DMed cohort, respectively. Next generation sequencing (NGS) assay (381/733 genes panel) was performed on tumor and plasma samples to capture tumor somatic single-nucleotide variants (SNVs). The gene variants information was summarized, analyzed and visualize using the maftools R package (Mayakonda et al. 2018.).

Results: In total, DNA alterations were detected in 93.86% (275/293) plasma samples and 90.91% (170/187) in tumor samples, which variant levels included pathogenic, likely pathogenic and variant of uncertain significance. The most commonly altered genes in ctDNA results were TP53 (52%), TERT (24%), LRP1B (22%), FAT1 (15%), CTNNB1 (14%). While in HCC tumor testing results were TP53 (55%), TERT (27%), CTNNB1 (17%), SPTA1 (14%) and ARID1A (12%). Both the most variant classification was missense, next were frameshift indel. Both the top five affected pathways were TP53, RTK-RAS, NOTCH, WNT and Cell Cycle. In WNT pathway related genes, the highest alteration frequency tumor oncogene was CTNNB1, and suppressor gene was APC. We analyzed mutually exclusive and co-occurring event on top 25 mutated genes. In ctDNA analysis results, the probability of co-mutation between LRP1B and KMT2A gene was the highest (p < 0.001), while mutual exclusion between ARID1A and EGFR gene was the highest.

Conclusion: Circulating tumor DNA mutation detection in HCC is feasible in the absence of HCC tissue DNA. The analysis results revealed the genomic landscape characteristics of blood-derived ctDNA in HCC were consistent with tumor tissues, which can provided HCC potential prognosis and treatment related genes information to further study.

Citation Format: Dinghua Yang, Sheng Yu, Baitang Guo, Tingting Chen. Genomic landscape analysis in tumor tissue and circulating tumor DNA of Chinese patients with hepatocellular carcinoma [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 5119.
Abstract 5120: Deciphering melanoma CTC signatures leading to immune escape and brain metastasis: The first MRI CTC xenograft model
Authors: Bowley Tetiana , Marchetti Dario
Published: June 2022

Melanoma brain metastasis (MBM) is a devastating disease which is diagnosed in up to 60% of melanoma patients and 80% of patients at autopsy. While microarray studies identified many genes differentially expressed between MBM and extracranial metastasis, magnetic resonance imaging (MRI) is the established technology to evaluate clinical MBM. Circulating Tumor Cells (CTCs) are “seeds” of fatal metastasis and smallest functional units of cancer. Defining mechanisms which regulate the spatial and temporal competence of patient-isolated CTCs driving MBM thus presents the opportunity to suppress its occurrence. Hypothesis is that MRI has fundamental translational relevance to validate preclinical CTC models and that MBM onset depends upon the absence or presence of human immune system cells (Hu-mice), and mice gender and age. We isolated CTCs from blood of primary/metastatic melanoma patients, and confirmed CTC detection by CellSearch࣪, Parsortix࣪, high-definition microscopy and RNA-Seq profiling. Second, we injected melanoma CTCs in immunodeficient but also immunocompetent xenografts (NBSGW female/male mice generated by multiple donors CD34+ cells), and assessed tumor progression by IVIS and MRI with parallel monitoring of CTC gene-specific transcriptional activation. Third, we discovered specific differences in number of CTCs/CTC clustering patterns and CTC signatures which were dependent upon MBM status. Quantitation of melanoma CTC-derived signatures enabled serial noninvasive monitoring of tumor burden. Lastly, we found that these differences and the occurrence, severity and spatial distribution of MBM highly depended from gender, age, and immunocompetency status of injected animals. Thus, by reporting the first MRI CTC xenograft model, its use for melanoma CTC interrogation can provide a platform for early monitoring of response to immunotherapeutic and/or targeted interventions in melanoma, and contribute to rational therapy applications, notably for MBM prediction or prevention.

Citation Format: Tetiana Bowley, Dario Marchetti. Deciphering melanoma CTC signatures leading to immune escape and brain metastasis: The first MRI CTC xenograft model [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2022; 2022 Apr 8-13. Philadelphia (PA): AACR; Cancer Res 2022;82(12_Suppl):Abstract nr 5120.
Abstract 513: Radiomic profiles revealing targets for therapy in cervical cancer
Authors: Halle Mari Kyllesø , Hodneland Erlend , Hoivik Erling , Wagner-Larsen Kari , Lura Njål G. , Dybvik Julie , Forsse David , Bertelsen Bjørn I. , Krakstad Camilla , Haldorsen Ingfrid S. , Bozickovic Olivera , Woie Kathrine
Published: June 2022

Magnetic resonance imaging (MRI) has emerged as an important part of the diagnostic work-up in cervical cancer, with tumor volume and extent as important prognosticators. MRI radiomic tumor features may aid in prognostication and tailoring of treatment in cervical cancer. We extracted whole-volume radiomic texture features from 124 manually segmented tumors and performed unsupervised clustering yielding two distinct clusters. Overlapping clinicopathologic, genomic (whole exome sequencing, n=61), transcriptomic (L1000 arrays, n= 65) and molecular biomarker (n=82) data were applied to characterize the clusters. Independent of tumor size parameters, patients in cluster II had significantly reduced disease-specific survival as compared to those in cluster I (p>0.001), also within squamous cell carcinomas (n=96, p<0.001). Cluster II associated with high age (p=0.02), high FIGO-2009 stage (p<0.001), high BMI (p=0.03) and PR negative tumors (p=0.004). Distinct mutational and copy-number profiles were detected for the two clusters. By gene set enrichment analyses, gene sets associated to immune activation were enriched in cluster I tumors, whilst gene sets associated with oxidative phosphorylation and epithelial to mesenchymal transition associated to cluster II tumors. This study links radiomic signatures to distinct genomic profiles that may potentially aid in prognostication and tailoring of treatments in cervical cancer patients.

Citation Format: Mari Kyllesø Halle, Erlend Hodneland, Erling Hoivik, Kari Wagner-Larsen, Njål G. Lura, Julie Dybvik, David Forsse, Bjørn I. Bertelsen, Camilla Krakstad, Ingfrid S. Haldorsen, Olivera Bozickovic, Kathrine Woie. Radiomic profiles revealing targets for therapy in cervical 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 513.
Abstract 5131: Deep learning-based tumor microenvironment cell types mapping from H&E images of lung adenocarcinoma using spatial transcriptomic data
Authors: Choi Hongyoon , Na Kwon Joong , Koh Jaemoon , Kim Young Tae
Published: June 2022

Purpose Spatial characterization of cell types of the tumor microenvironment (TME) is a key to finding new targets as well as developing biomarkers for immuno-oncology treatment. Here, we develop and validate a deep learning model trained by integrating H&E images of lung adenocarcinoma and spatial transcriptomic data to decipher spatial mapping of multiple cell types in TME only using H&E images.

Methods A total of 21 H&E images combined with spatial transcriptomic data of lung adenocarcinoma were used to develop a model. As each spot of spatial transcriptomic data consisted of a few cells, the cell types were inferred by CellDART, a domain adaptation-based method to estimating five cell types defined by single cell RNA-seq (scRNA-seq) data - B, T/NK, myeloid cells, fibroblasts, and epithelial cells. A convolutional neural network using H&E image patches as inputs was developed to predict the cell type scores for each corresponding spot. For the external validation, the model estimated the cell type scores from H&E-stained tissue image patches of lung adenocarcinoma of the Cancer Genome Atlas (TCGA-LUAD). Furthermore, the model inferred cell types from H&E images of lung adenocarcinoma tissues acquired by different patients and the results were compared with immunohistochemistry.

Results The cell types inferred by the model using H&E image patches were significantly correlated with those derived by spatial transcriptomic data as an internal validation. The mean value of the deep learning-based cell types scores estimated by the TCGA-LUAD tissue images was significantly correlated with the cell type scores estimated by bulk RNA-seq data of corresponding TCGA data. When cell type inference maps of independent lung adenocarcinoma H&E images were spatially co-registered with immunohistochemistry slides, including CD3, CD20, and CD68, pixel-wise correlation analysis revealed positive significant correlations.

Conclusions A deep learning model to infer spatial cell types of tumor microenvironment using H&E images was developed. The results of cell type scores were positively correlated with bulk RNA-seq-based immune cell enrichment scores and pixelwise analyses on immunohistochemistry images using typical cell type markers. This approach can provide objective and flexible deep learning-based models for characterizing tumor microenvironment.

Citation Format: Hongyoon Choi, Kwon Joong Na, Jaemoon Koh, Young Tae Kim. Deep learning-based tumor microenvironment cell types mapping from H&E images of lung adenocarcinoma using spatial transcriptomic data [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 5131.
Abstract 5136: Development of a novel gene expression signature that correlates with intratumor microvessel density and antitumor activity of lenvatinib
Authors: Kuronishi Megumi , Ozawa Yoichi , Kimura Takayuki , Kato Yu
Published: June 2022

Lenvatinib is a multiple receptor tyrosine kinase inhibitor and an angiogenesis inhibitor targeting mainly vascular endothelial growth factor (VEGF) and fibroblast growth factor (FGF) receptors. In our previous study, we reported that human xenograft tumors sensitive to lenvatinib showed higher microvessel density (“MVD (IHC)”) than relatively resistant tumors in immunohistochemistry (IHC). Although MVD (IHC) is a potential indicator to predict antitumor activity of lenvatinib, available IHC data are limited. Here, we investigated a novel gene expression signature (MVD gene score) calculated from RNA-sequencing (RNA-seq) data that reflects MVD (IHC) and antitumor effect of lenvatinib.

Tumor tissues were collected from 12 mouse syngeneic tumor models followed by RNA-seq. We also stained CD31 in the same tumors and calculated MVD (IHC) score by dividing the number of CD31 positive blood vessels by tumor area. Genes that met the following criteria were selected for MVD gene score calculation: (1) genes that are specifically expressed on endothelial cell cluster in in-house single cell RNA-seq data using Hepa 1-6 mouse hepatocellular carcinoma tumor tissues; (2) well-known angiogenesis markers from literature or genes whose expression show significant correlation with MVD (IHC) in mouse syngeneic tumor models. MVD gene score was calculated as the average of logarithm transformed gene expression values. In order to examine applicability of MVD gene score, we compared MVD (IHC) and MVD gene score to lenvatinib antitumor activities on mouse syngeneic tumors.

We measured MVD (IHC) and computed MVD gene score from RNA-seq data using matched tumor tissues of 12 mouse syngeneic models. MVD gene score was correlated with MVD (IHC). As reported, lenvatinib sensitive tumor models showed higher MVD (IHC) score. They also showed higher MVD gene score calculated from RNA-seq data.

Our newly developed MVD gene score has a potential to predict the antitumor activity of angiogenic inhibitors such as lenvatinib using RNA-seq data in tumor samples. Further validation study is ongoing using various human tumor samples.

Citation Format: Megumi Kuronishi, Yoichi Ozawa, Takayuki Kimura, Yu Kato. Development of a novel gene expression signature that correlates with intratumor microvessel density and antitumor activity of lenvatinib [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 5136.