bluuebunny/crossref_metadata_2025 huggingface dataset:
Abstract 5089: Small molecule Bax activator enhances immunotherapy against lung cancer
Authors: Anisuzzaman Abu Syed Md , Talukdar Muhammad Waliul , Deng Xingming
Published: April 2023

Induction of apoptosis is a critical mechanism underlying the durable efficacy of cancer treatment agents. Bax is a major proapoptotic protein whose activation is required for apoptotic cell death. We have recently discovered that small molecule Bax activator CYD-2-11 targets the S184 structural pocket in the c-terminal tail of Bax, directly activates its proapoptotic activity via conformational change and formation of Bax homo-oligomers in mitochondrial membranes. CYD-2-11 has potent anti-tumor activity against lung cancer in various animal models. Here we found that CYD-2-11 in combination with radiation, cisplatin or BH3 mimetic Bcl2 inhibitor venetoclax (ABT-199) synergistically suppressed tumor growth in non-small cell lung cancer (NSCLC) xenograft models. It is known that LKB1 mutations or deficiency are associated with therapeutic resistance to PD-1/PD-L1 checkpoint blockade immunotherapy. To test whether Bax activator CYD-2-11 reverses resistance to immunotherapy in immunologically “cold” genetically engineered LSL-KRASG12D LKB1 fl/fl (KL) mouse model, KL mice were treated with CYD-2-11, PD-L1 antibody or in combination. Results indicate that PD-L1 antibody alone has no significant anti-tumor effect in KL mice. CYD-2-11 alone can reduce tumor burden, but combination of CYD-2-11 with PD-L1 antibody has very strong synergistic effect and produces maximum efficacy in tumor burden reduction and significantly prolongs survival of KL mice. Mechanistically, combined treatment with CYD-2-11 and PD-L1 antibody enhances T cell infiltration in tumor tissues. These findings provide preclinical evidence for pharmacologic combinations of small molecule Bax activator with chemoradiotherapy, Bcl-2 inhibitor or PD-L1 antibody as novel strategies to treat lung cancer more effectively.

Citation Format: Abu Syed Md Anisuzzaman, Muhammad Waliul Talukdar, Xingming Deng. Small molecule Bax activator enhances immunotherapy against lung 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 5089.
Abstract 5202: NCG-M humanized mice: an excellent model for human immune reconstitution of myeloid lineages
Authors: Wang Huiyi , Fan Jialu , Guo Shiying , Zhang Mingkun , Li Shuai , Chen Santi Suryani , Li Zhiying , Moore Mark Wade , Zhao Jing , Gao Xiang , Ju Cunxiang
Published: April 2023

Severe immunodeficient mice engrafted with human hematopoietic stem cells (HSC) have been extensively used in immuno-oncology related studies to evaluate the efficacy of cancer therapies. However, because of species differences, murine cytokines provide limited support to human immune cells, thus immunodeficient mice have limited engraftment of human immune cells. Increasing evidence has shown that myeloid cells, especially macrophages and dendritic cells, are critical for the induction of anti-tumor immunity. We developed a mouse model, NCG-M, that can support human T, B, NK and myeloid cells such that in vivo evaluation of agents that require the interplay between these immune cells can be examined. This model was genetically engineered on the severe immunodeficient strain NCG and can produce human granulocyte/macrophage colony stimulating factor 2 (GM-CSF, also known as CSF2), interleukin-3 (IL-3) and stem cell factor (SCF, also known as KITLG). Upon human CD34+ HSC cell engraftment, increased myeloid lineage cells, such as granulocytes, monocytes, neutrophils, macrophages, and dendritic cells, were observed in the NCG-M cohort compared to NCG mice. The NCG-M mouse also supports the development of human T cells, and preliminary data showed increased B and NK cells. The NCG-M is an appropriate mouse model for studying the efficacy of therapeutic agents that require human T cells and myeloid cells.

Citation Format: Huiyi Wang, Jialu Fan, Shiying Guo, Mingkun Zhang, Shuai Li, Santi Suryani Chen, Zhiying Li, Mark Wade Moore, Jing Zhao, Xiang Gao, Cunxiang Ju. NCG-M humanized mice: an excellent model for human immune reconstitution of myeloid lineages. [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 5202.
Abstract 5206: A novel mouse model for preclinical efficacy evaluation of anti-CD3/GPRC5D bispecific antibodies
Authors: Yang Xiaoliu , Liang Xinyuan , Zhang Mingkun , Chen Santi Suryani , Li Zhiying , Trujillo Erica , Moore Mark Wade , Zhao Jing , Gao Xiang , Ju Cunxiang
Published: April 2023

The FDA has granted Breakthrough Therapy Designation (BTD) for talquetamab, which targets both GPRC5D on multiple myeloma cells and CD3 on T-cells, for treating adult patients with relapsed or refractory multiple myeloma. Results showed that patients who received talquetamab experienced adverse events, including cytokine release syndrome (CRS), neutropenia, skin-related adverse events and dysgeusia. To allow preclinical in vivo evaluation of anti-human CD3/GPRC5D bispecific antibodies, we developed a BALB/c-hCD3EDG mouse model and a human GPRC5D-expressing A20 murine lymphoma cell line. The BALB/c-hCD3EDG cohort (6-8 weeks old) was subcutaneously inoculated with A20-hGPRC5D cells and then treated with anti-CD3/GPRC5D bispecific antibodies. As expected, anti-CD3/GPRC5D bispecific antibody treatment strongly inhibited the growth of A20-hGPRC5D tumors in BALB/c-hCD3EDG mice.

To allow a comprehensive safety evaluation of anti-CD3/GPRC5D candidates, including the possibility of CRS and target organ toxicity, we engineered BALB/c-hCD3EDG mice that express hGPRC5D, denoted BALB/c-hCD3EDG/hGPRC5D. Human GPRC5D was detected in the liver, lung, thymus, adipose tissue, and thyroid. In vivo evaluation of anti-CD3/GPRC5D therapy in this mice is currently underway.

In conclusion, BALB/c-hCD3EDG/hGPRC5D and BALB/c-hCD3EDG mice bearing A20-hGPRC5D tumors are suitable models for preclinical studies testing anti-CD3/GPRC5D bispecific antibodies.

Citation Format: Xiaoliu Yang, Xinyuan Liang, Mingkun Zhang, Santi Suryani Chen, Zhiying Li, Erica Trujillo, Mark Wade Moore, Jing Zhao, Xiang Gao, Cunxiang Ju. A novel mouse model for preclinical efficacy evaluation of anti-CD3/GPRC5D bispecific antibodies. [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 5206.
Abstract 5223: Circulating lipoprotein lipids and colorectal cancer risk: A Mendelian randomization analysis from the GECCO consortium
Authors: Liu Lili , Wen Wanqing , Long Jirong , Assimes Themistocles L , Bujanda Luis , Gruber Stephen B , Küry Sébastien , Lynch Brigid , Qu Conghui , Thomas Minta , White Emily , Woods Michael O. , Peters Ulrike , Zheng Wei
Published: April 2023

Background Conventional observational studies have reported conflicting results regarding the association between low density lipoprotein cholesterol (LDL-C) and risk of colorectal cancer (CRC). We conducted a Mendelian randomization analysis to address this association.

Methods Single-nucleotide polymorphisms (SNPs) associated with five blood lipids (total cholesterol, HDL-C, high-density lipoprotein cholesterol [HDL-C], non-HDL-C, and triglyceride) were obtained from a genome-wide association study (GWAS) meta-analysis of European ancestry in the Global Lipids Genetics Consortium (GLGC, N ≤ 1 319 982), and two lipids (apolipoprotein A1 and apolipoprotein B) from a GWAS in the UK Biobank (UKB, N ≤ 441 016). Summary statistics were obtained for these SNPs from a GWAS of CRC in the Genetics and Epidemiology of Colorectal Cancer Consortium (GECCO) including 34,869 cases and 29,051 controls. Associations with CRC risk per one standard deviation increase in the genetically predicted lipids level were generated using inverse-variance weighted random-effects models.

Results No overall association was observed between genetically predicted levels of blood lipids and CRC risk. However, increased risks were observed for all LDL-C related traits among women, including total cholesterol (OR = 1.10; 95%CI = 1.02, 1.18), LDL-C (1.07; 1.00, 1.14), non-HDL-C (1.09; 1.02, 1.16), and apolipoprotein B (1.11; 1.02, 1.21); whereas no significant association was found among men. Similar but ostensibly stronger associations of these traits

were seen with distal colon cancer cases, with no significant association showing on proximal colon or rectum cancer cases. We also observed similar positive associations of LDL-C related traits among those having CRC before their 50 years. Of note, risk reduction was found for apolipoprotein A1, a major component of HDL-C, in these early-onset cases (0.87; 0.78, 0.98).

Conclusion Results from this study suggest that high circulating LDL-C levels may increase the risk of CRC, particularly cancer of the distal colon, and the association may differ by sex and age at CRC onset.

Key words: Blood lipids; colorectal cancer; Mendelian randomization.

Citation Format: Lili Liu, Wanqing Wen, Jirong Long, Themistocles L Assimes, Luis Bujanda, Stephen B Gruber, Sébastien Küry, Brigid Lynch, Conghui Qu, Minta Thomas, Emily White, Michael O. Woods, Ulrike Peters, Wei Zheng. Circulating lipoprotein lipids and colorectal cancer risk: A Mendelian randomization analysis from the GECCO consortium. [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 5223.
Abstract 5252: Salsolinol suppresses the STAT3-EMT axis in liver carcinogenesis
Authors: Oh Hyeon Jeong , Park Seah , Yang Hong-Kyung , Ryu Hoon , Surh Young-Joon , Kim Dae-Yong , Na Hye-Kyung
Published: April 2023

Liver cancer is one of the most common malignancies and a leading cause of death worldwide. However, it is still very difficult to treat and prevent liver cancer. Salsolinol (SAL) is an endogenous catechol isoquinoline generated by the condensation of dopamine with acetaldehyde, a major metabolite of ethanol. In the present study, we found that SAL inhibited the phosphorylation, dimerization, and subsequent nuclear translocation of STAT3 in a concentration-dependent manner in SK-Hep1 cells, a hepatic carcinoma cell line. SAL suppressed the expression of Cdk 4 and Cyclin D1, the major target protein of STAT3, and induced the cell cycle arrest at Go/G1 phase. However, SAL induced the expression of STAT1 in SK-Hep1 cells. In addition, SAL enhanced the expression of the cell cycle regulators such as p53 and p21 in SK-Hep1 cells. Epithelial-mesenchymal transition (EMT) promotes tumor invasion and metastasis in malignancies through STAT3 activation. SAL induced the expression of E-cadherin while suppressing the expression of Snail, N-cadherin, and Slug, which are related to EMT. SAL inhibited the migration of SK-Hep1 cells by suppressing the expression of MMP-9 and MMP-2. Furthermore, intraperitoneal injection of SAL reduced the number of tumors in the diethylnitrosamine (DEN)-induced liver carcinogenesis model without affecting the body weight change. Proliferative markers PCNA and KI-67 and p-STAT3 were down-regulated in the SAL-injected mice liver cancer tissue. The plasma level of alpha-fetoprotein, a tumor marker in the blood, was also reduced in the SAL-injected mice. Interestingly, SAL injection significantly reduced the anxiety-like behavior without affecting the locomotive activities in the DEN-induced hepatocellular carcinoma mouse model. Moreover, SAL injection recovered tyrosine hydroxylase, a key enzyme for producing dopamine from tyrosine in the substantia nigra compacta of DEN-treated mice. Together, SAL inhibits STAT3 signaling, thereby suppressing the cell cycle progression and EMT in SK-Hep1 cells, which may account for its anti-depressant and anti-carcinogenic activity in DEN-induced liver carcinogenesis.

Citation Format: Hyeon Jeong Oh, Seah Park, Hong-Kyung Yang, Hoon Ryu, Young-Joon Surh, Dae-Yong Kim, Hye-Kyung Na. Salsolinol suppresses the STAT3-EMT axis in liver carcinogenesis. [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 5252.
Abstract 5313: The expression of phospho-Rb S249, N-cadherin, E-cadherin, and B-catenin as a prostate proteomic score for the risk classification of prostate cancer tumors
Authors: Cortes Sheila M. Valle , Cardona Pedro Santiago , Osterman Carlos Diaz , Morales Jaileene Perez , Maldonado Darielys Maldonado , Deya Gilberto Ruiz
Published: April 2023

Prostate cancer (PCa) ranks as the second leading cause of cancer-related deaths in American men. Most PCa deaths occur due to aggressive, fast-growing tumors that develop into metastasis when cancer cells spread to other body parts, including the lymph nodes, bones, etc. However, men could also be affected by indolent, slow-growing tumors, which may exist for an extended period without causing any symptoms or death. Current prognostic methods, such as Prolaris, or Oncotype, do not precisely specify how aggressive cancer might be, resulting in overtreating patients with indolent PCa. Therefore, improved ways to differentiate indolent from aggressive tumors are needed. In the presented study, we have validated the clinical potential of phospho-Rb S249, N-cadherin, B-catenin, and E-cadherin to discriminate between indolent and aggressive PCa tumors. We hypothesize that the biomarkers phospho-Rb S249, N-cadherin, E-cadherin, and B-catenin expression in PCa will be a clinical tool for the risk classification of PCa tumors when used as part of a prostate proteomic (PPS) that can predict the risk of the tumor of evolving into an aggressive phenotype. We conducted immunohistochemistry staining of these biomarkers on PCa tumor microarrays and correlated it with the patient’s clinical parameters, such as tumor size, lymph node invasion, metastasis, etc. Also, we correlated it with the PSA levels and created a PPS. The combination of our proteomic biomarkers predicts patient staging by 53.83%. Adding clinical data such as the tumor size, lymph node invasion, and metastasis (TNM) increases the precision of the prediction to 83.21%. In addition, the PSA levels increased as the phospho-Rb S249 expression and N-cadherin rise, accompanied by up-regulation of E-cadherin. Also, the PPS showed to distinguish between normal prostate tissue, hyperplasia, and PCa. Results suggest that these biomarkers could serve as a tool for the risk classification of indolent and aggressive PCa tumors.

Citation Format: Sheila M. Valle Cortes, Pedro Santiago Cardona, Carlos Diaz Osterman, Jaileene Perez Morales, Darielys Maldonado Maldonado, Gilberto Ruiz Deya. The expression of phospho-Rb S249, N-cadherin, E-cadherin, and B-catenin as a prostate proteomic score for the risk classification of prostate cancer tumors. [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 5313.
Abstract 5413: AIVE, an artificial intelligence protein miner based on omics data to discover new cancer druggable candidates
Authors: Hong Dongwan
Published: April 2023

Recently, AI software including Alphafold, RoseTTAFold and ESMFold, can predict protein structures based on amino acid sequences, and DeepMind (Alphafold2) and Meta (ESMFold) opened hundreds of millions of protein structures to world-wide clinical researchers. These tools are possible to generate protein structures in a shorter time and less cost than traditional protein study methods. Finally, they can lead to the advent of the protein rush generation. To evaluate accurately and quickly the prediction of structures of mutations embedding in protein sequences, we developed an Artificial Intelligence analytics toolkit for predicting Virus mutation in protEin (AIVE; http://ai-ve.org) that clinical researchers can use easily on web. An amino acid property eigen selection score (APESS) defined in AIVE is a comprehensive mathematical analysis model, andit was calculated basically by below: 1 artificial intelligence base clustering (AIBC) using predicted aligned error (PAE) and per-residue confidence (pLDDT) values, 2 base level change rate (BLCR)using the probability of occurrence of mutation in each amino acid sequence or nucleotide sequence, 3chemical property eigen score (CPES) using amino acid properties (PH, Residue and Hydrophobic),and 4polarity based feature selection (PBFS) using continuous position of polar amino acids including S, C, N, Q, T, and Y [APESS = AIBC ´ BLCR ´ CPES ´ PBFS]. AIVE also calculated the docking score using AlphaFold to estimate protein to protein interaction (PPI) among hosts (or mutations) and provided a graph as final report showing APESS distribution among mutation group, and evaluation of APESS and docking score. For example, according to APESS score in lung cancer, we can find out the feasibility of structure transformation of H-bond in EGFR protein (L858R) tyrosine kinase and also show the differences of tyrosine kinase affinity between each lung cancer patients. However, for the T790M mutation of EGFR protein, there was no significant difference compared with the wild type (790). The execution time of AIVE (GPU based web server platform) depends on the length of the input sequence, and it completes such an example case in an average of 2-3 hours. AIVE can suggest anti-cancer candidates in protein-level and present a precision medicine model which is available to implement close to real time.

Citation Format: Dongwan Hong. AIVE, an artificial intelligence protein miner based on omics data to discover new cancer druggable candidates. [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 5413.
Abstract 5421: Constructing the largest-scale knowledge graph using all PubMed abstracts and its application for highly specific and accurate knowledge retrieval
Authors: Sui Xin , Zhang Yuan , Pan Feng , Sun Donghu , Chung Menghan , Zhang Jinfeng
Published: April 2023

Most of the biomedical knowledge the research community has acquired during the past few decades has been deposited in scientific literature as unstructured text. Converting the unstructured text into the structured form will enable novel methodologies and applications for scientific discovery that can fully harness the power of the existing knowledge. To this end, two fundamental questions need to be addressed: named entity recognition (NER) and relation extraction (RE). NER deals with identifying the concepts or entities in texts, such as diseases, genes/proteins, chemical compounds, etc. while RE aims to extract the relations among these entities. Together, the extracted information forms a knowledge graph (KG) where the nodes are entities in the texts and the edges represent their relationships. KGs can link concepts within existing research to allow researchers to find connections that may have been difficult to discover without them. The LitCoin Natural Language Processing (NLP) Challenge was recently organized by NCATS of NIH and NASA to spur innovation by rewarding the most creative and high-impact uses of biomedical text to create KGs. Our team participated in the challenge and ranked first place. We have applied the methods we developed for the LitCoin NLP challenge to all PubMed abstracts and constructed the largest-scale biomedical KG to date. We show that powerful and versatile query functions can be implemented on top of the KG to enable highly specific and accurate knowledge retrieval and inference of causal and indirect relationships.

Citation Format: Xin Sui, Yuan Zhang, Feng Pan, Donghu Sun, Menghan Chung, Jinfeng Zhang. Constructing the largest-scale knowledge graph using all PubMed abstracts and its application for highly specific and accurate knowledge retrieval. [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 5421.
Abstract 5490: Combination of the PI3k inhibitor Paxalisib with the nucleoside analog Gemcitabine over-activates the integrated stress response and induces atypical teratoid/rhabdoid tumor cell death
Authors: Findlay Tyler , Malebranche Kristen , Geethadevi Anupa , Raabe Eric , Eberhart Charles , Rubens Jeffrey
Published: April 2023

We have previously identified high activation of both mTORC1 and mTORC2 in the aggressive infantile brain tumor, atypical teratoid/rhabdoid tumor (AT/RT). Paxalisib is a highly brain penetrant PI3k inhibitor which acts upstream of mTOR to simultaneously inhibit mTORC1/2. Paxalisib is well-tolerated in ongoing pediatric clinical trials. In murine orthotopic xenograft models of AT/RT, Paxalisib significantly extended survival (CHLA-06: 40 to 54 days, p=0.001; BT12: 21 to 35 days, p=0.02). However, to improve the durability of this single agent therapy, we evaluated the efficacy of rational therapeutic partners. Analysis of RNAseq following mTOR inhibition (mTORi) of 4 cell lines representative of AT/RT showed an upregulation of genes associated with the integrated stress response (ISR), (ATF4: CHLA-06, t-test p<0.05; CHOP: CHLA-02, CHLA-06 t-test p<0.001). The activation of the ISR is further suggested through induction of PPP1R15A, a gene whose activation is closely associated with stress events and leads to downstream cell growth arrest and apoptosis (BT37 t-test p<0.001, CHLA-02, CHLA-06, CHLA-266, t-test p<0.01). Gemcitabine is a pyrimidine nucleoside prodrug, which inhibits DNA synthesis and induces DNA damage. As a result, Gemcitabine also induces phosphorylation of eIF2α and the activation of the ISR. While the ISR can be protective, intense or prolonged activation leads to cell death. We therefore hypothesized that Paxalisib would combine with Gemcitabine to hyper-activate the ISR and drive AT/RT cell death. Through Western blot analysis, we demonstrate that combination therapy increases phospho-eIF2α, ATF4, and CHOP compared to DMSO control. Furthermore, we demonstrate that combination therapy increases apoptosis (Western blot cPARP, MUSE Annexin V Assay) and synergizes to decrease AT/RT cell growth (SynergyFinder BLISS score CHLA-06: 16.8, BT12: 14.3, CHLA-266: 13.9, BT37: 11.4, CHLA-05: 10.4). Ongoing treatment of mice bearing CHLA06 orthotopic tumors demonstrate that combination therapy further extends median survival. Our studies suggest that pharmacologically activating the ISR may be an effective strategy to target AT/RT and combination of Paxalisib and Gemcitabine may be an effective treatment to help extend survival in this deadly disease.

Citation Format: Tyler Findlay, Kristen Malebranche, Anupa Geethadevi, Eric Raabe, Charles Eberhart, Jeffrey Rubens. Combination of the PI3k inhibitor Paxalisib with the nucleoside analog Gemcitabine over-activates the integrated stress response and induces atypical teratoid/rhabdoid tumor cell death. [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 5490.
Abstract 5495: Combination therapy with poly (ADP-ribose) polymerase inhibitor in endometrial cancer
Authors: Xu Conway , Hou Xiaonan , Jessen Erik , Wang Chen , Weroha S. John
Published: April 2023

Objective: Endometrial cancer (EC) is the most common gynecologic malignancy and serous histology is the second most common subtype. It is associated with poorer outcomes as recurrent disease tends to be resistant to chemotherapy. Given the dearth of proven targeted therapies in this setting and the tendency for serous ECs to have homologous recombination deficiency (HRD), this study aimed to test the synergy between poly (ADP-ribose) polymerase inhibitors (PARPis) and SN38, an active metabolite of DNA topoisomerase I, irinotecan. PARPis are currently FDA approved in ovarian cancer and have their greatest efficacy in tumors with HRD while topoisomerase inhibitors are a standard chemotherapy option for recurrent cancer.

Methods: A genomic instability score (GIS) was derived from low-pass whole genome sequencing-based bioinformatics analysis according to three previously reported HRD metrics: telomeric allelic imbalance, large state transitions, and loss of heterozygosity. GIS was calculated for 34 patient derived xenograft (PDX) tumors, of which eight had serous histology. PARPi, rucaparib, and SN38 were used to treat EC PDX tumors in ex vivo 3D cell culture experiments using RealTime Glo for viability after 4-5 days of drug exposure. Synergy was assessed by calculating the combination index (CI) using Chou-Talalay method.

Results: GIS ≥ 42 was noted in 62.5% (5/8) of serous EC PDX tumors and only 19.2% (5/26) of non-serous EC PDX tumors. Synergy (CI < 1.0) between rucaparib and SN38 was demonstrated in 85.7% (6/7) of serous EC PDX ex vivo 3D cell culture experiments, but only 62.5% (10/16) of non-serous EC PDX tumors.

Conclusions: Most serous EC PDX tumors had a high GIS, consistent with HRD, when compared to a minority of non-serous histologies. Furthermore, combination therapy of rucaparib and SN38 had a synergistic effect in almost all serous EC PDX models, while still being effective in a majority of non-serous models. Additional PDX tumors need to be assessed to establish the predictive value of a GIS on synergy between rucaparib and SN38. Further in vivo studies are also needed to confirm these ex vivo findings of synergy. Ultimately, the hope is to generate preclinical data which justifies the use of PARPis in serous ECs as monotherapy or in combination with other anticancer therapies.

Citation Format: Conway Xu, Xiaonan Hou, Erik Jessen, Chen Wang, S. John Weroha. Combination therapy with poly (ADP-ribose) polymerase inhibitor in endometrial 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 5495.
Abstract 5747: A novel combination therapy of arenavirus vectors and PD1-IL2v strongly potentiates tumor specific T cell responses resulting in synergistic anti-tumor efficacy
Authors: Strauss Judith , Reckendorfer Diana , Pojar Kimberly , Deak Laura Codarri , Nicolini Valeria , Sutmuller Roger , Klein Christian , Umaña Pablo , Orlinger Klaus K. , Lauterbach Henning
Published: April 2023

It is undisputed that CD8 T cells play a critical role in controlling tumor growth and killing tumor cells, and that tumor specific T cells are essential for clinical responses to immune checkpoint inhibitors. Yet, the induction and maintenance of functional tumor antigen specific T cell responses represent still one of the greatest challenges in cancer immunotherapy.

Therapeutic cancer vaccines based on engineered arenavirus vectors have been shown preclinically to overcome self-tolerance to tumor associated antigens (TAA) and mount unprecedented T cell responses, both, in preclinical tumor models as well as HPV16+ cancer patients in an ongoing Phase 1/2 trial. The novel PD-1 targeted IL-2 variant (PD1-IL2v) was created to maximize the potency of IL-2R activation of effector T cells while overcoming the toxicities of wildtype IL-2. Cis binding of PD1-IL2v to PD-1 and IL-2Rβγ on the same cell was recently shown to promote differentiation of stem-like CD8 T cells into better effectors and was therefore identified as an ideal compound for combination therapies with arenavirus vectors. Combination of vectors encoding tumor associated foreign or self-antigens with PD1-IL2v lead to a massive, up to 60-fold, increase of peripheral tumor specific CD8 T cell numbers compared to vector treatment alone. Notably, frequency of peripheral regulatory T cells was not enhanced by this combination. The massive increase of TAA specific CD8 T cells was associated with synergistic anti-tumor efficacy in two independent tumor models, resulting in longer survival times and up to 60% complete responders. Complete tumor clearance resulted in a stable memory T cell population and protection against tumor re-challenge. Ongoing studies will address modifications of the tumor microenvironment and further characterization of tumor infiltrating T cells upon combination therapy.

These preclinical data establish the combination of arenavirus vectors and PD1-IL2v as a promising next generation cancer immunotherapy.

Citation Format: Judith Strauss, Diana Reckendorfer, Kimberly Pojar, Laura Codarri Deak, Valeria Nicolini, Roger Sutmuller, Christian Klein, Pablo Umaña, Klaus K. Orlinger, Henning Lauterbach. A novel combination therapy of arenavirus vectors and PD1-IL2v strongly potentiates tumor specific T cell responses resulting in synergistic anti-tumor efficacy. [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 5747.
Abstract 5860: ZEB1-driven reconstruction of the hyaluronan network establishes a pro-metastatic microenvironment in lung adenocarcinoma
Authors: Lee Sieun , Park Jihye , Kim Eun Ju , Park Sungsoo , Kurie Jonathan M. , Ahn Young-Ho
Published: April 2023

Hyaluronan, one of the major components of the extracellular matrix (ECM), surrounds and protects neighboring cells from diverse stresses. Hyaluronan promotes epithelial-to-mesenchymal transition (EMT) and metastasis in various epithelial cancers. However, the mechanism by which the hyaluronan network built up by cancer cells regulates cancer progression and metastasis in the tumor microenvironment has not been elucidated in detail. In this study, ITIH2, a hyaluronan-binding protein that participates in building a hyaluronan network, was confirmed to be secreted from mesenchymal-like lung cancer cells when co-cultured with cancer-associated fibroblasts. ITIH2 was transcriptionally upregulated by ZEB1, an EMT-inducing transcription factor. Hyaluronic acid synthase-2 (HAS2) was also up-regulated by ZEB1, and the expression of HAS2 and ZEB1 showed a positive correlation in the TCGA database. In addition, ZEB1 controlled alternative splicing and isoform expression of CD44, a hyaluronan receptor. Interestingly, ITIH2 knockdown and 4-methylumbelliferone (4-MU), a HAS inhibitor, reduced the formation of hyaluronan cables and inhibited the migration and invasion of lung cancer cells. Likewise, when lung cancer cells were treated with “Compound S,” an ITIH2 inhibitor predicted by a deep learning-based drug-target interaction algorithm, the formation of hyaluronan cables and cell migration were suppressed. In addition, the ITIH2 inhibitor prevented the growth of metastatic tumors in an orthotopic lung cancer mouse model. Collectively, the EMT signal controlled by ZEB1 establishes the pro-metastatic hyaluronan network in the tumor microenvironment, suggesting a novel strategy for targeting the hyaluronan network to suppress lung cancer progression and metastasis.

Citation Format: Sieun Lee, Jihye Park, Eun Ju Kim, Sungsoo Park, Jonathan M. Kurie, Young-Ho Ahn. ZEB1-driven reconstruction of the hyaluronan network establishes a pro-metastatic microenvironment in lung 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 5860.
Abstract 5869: The concordant disruption of B7/CD28 immune regulators predicts the prognosis of oral carcinoma
Authors: Chang Sih-Rou
Published: April 2023

Immune-escape is a critical factor determining the survival of malignancies including oral squamous cell carcinoma (OSCC). The immune escape of cancer cells may be driven by B7/CD28 family members and others on the tumor cells that form ligand-inhibitory receptor complexes with immune cells. Since the family members of B7/CD28 can functionally compensate, the concomitant disruption of multiple B7/CD28 family members in OSCC pathogenesis remains elusive. RNAseq performed on OSCC and paired oral mucosa retrieved the transcriptome of OSCC, and algorithms signified the general alterations of T cell population and the increase of macrophage population in tumors relative to controls. Upregulation of CD80, CD86, PD-L1, PD-L2, CD276 and CTLA4, and downregulation of L-ICOS in OSCC relative to normal mucosa was noted. In addition, there was a high concordance in the expression profile across the dysregulated regulators. The co-upregulation of CD276 and CTLA4 was associated with the decrease of T helper and NK cell population, and the increase of Treg and myeloid dendritic cell population. Interestingly, tumors harboring higher PD-L1 and/or PD-L2 expression accompany with lower ICOS or PD1 expression exhibited worse prognosis. In combination with these immunity features in primary tumors, the survival of patients with lymph node metastasis was further worsened. In the isografic models of murine OSCC cell lines, the decrease of CD4+T population in the lymph node and increase of myeloid dendritic cell population in the spleen were induced in the host mice. Many miRNAs are involved in the modulation of immune escape for malignancies. With the treatment of inhibitors against oncogenic miRNAs which enabled the OSCC suppression, the inhibitors of miR-146a and miR-211 also resulted in the down-regulation in the protein expression of multiple B7/CD28 members. This study concludes the eminent co-disruption of B7/CD28 members in OSCC tumors, which would confound the patient survival. The efficacy of miRNA inhibitors in abrogating oncogenicity and B7/CD28 members could be a therapeutic strategy.

Citation Format: Sih-Rou Chang. The concordant disruption of B7/CD28 immune regulators predicts the prognosis of oral carcinoma. [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 5869.
Abstract 5876: Sorafenib and Lenvatinib induce vascular responses in patient derived HCC on-Chip models
Authors: Mocellin Orsola , Robinson Abbie , Olczyk Aleksandra , Treillard Stephane , Olivier Thomas , Ng Chee , Heijmans Jeroen , Goubert Désiréé , Stok Arthur , van Tienderen Gilles , Verstegen Monique , Trietsch Sebastian , Lanz Henriëtte , Vulto Paul , Joore Jos , Queiroz Karla S.
Published: April 2023

Hepatocellular carcinoma (HCC) is the most common type of liver cancer. Its incidence is increasing, and is closely related to advanced liver disease. Interactions in the HCC microenvironment between tumor cells and the associated stroma actively regulate tumor initiation, progression, metastasis, and therapy response. In the present study, we used the OrganoPlate graft to establish a co-culture system consisting of dissociated HCC tumors (HCC 1-8) and cell lines, HCC derived fibroblasts and vasculature. Cultures were prepared and validated by assessing their response to Sorafenib and Lenvatinib (72 hours). Cultures had their viability (alamar blue assay), and chemokine/cytokine levels in the supernatant (Luminex) determined. In addition, the organization of the vasculature in the tumor compartment was studied through immunostainings, confocal imaging, and subsequent morphological analyses. HCC models were characterized by a range of specific markers, tumor (albumin), endothelial (CD31 and VE-Cadherin) and stromal (aSMA) cells. CD31 immunostained cultures were imaged, and morphology changes quantified. Sorafenib and Lenvatinib induced changes in the tumor vasculature area and organization. Hereby, we present vascularized patient-derived HCC models that include relevant cellular players of the HCC microenvironment. These co-cultures are highly suitable for studying specific cell types as well as patient-specific responses. We envision that this patient derived model will evolve to become a platform for understanding the interplay between angiogenesis, stroma and immune infiltrate in HCC.

Citation Format: Orsola Mocellin, Abbie Robinson, Aleksandra Olczyk, Stephane Treillard, Thomas Olivier, Chee Ng, Jeroen Heijmans, Désiréé Goubert, Arthur Stok, Gilles van Tienderen, Monique Verstegen, Sebastian Trietsch, Henriëtte Lanz, Paul Vulto, Jos Joore, Karla S. Queiroz. Sorafenib and Lenvatinib induce vascular responses in patient derived HCC on-Chip models [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 5876.
Abstract 5975: Discovery of mechanisms that modulate the hippo pathway and CDK6 expression in CDK4/6 inhibitor resistant breast cancer cells
Authors: Yu Chung-Jen , Zhuang Yong-Ji , Lin Ting-Yi , Chao Ta-Chung , Liu Chun-Yu , Tseng Ling-Ming , Lai Jiun-I
Published: April 2023

Hormone positive (HR+) breast cancer is the most common subtype in this population. The current standard of care in advanced HR+ breast cancer includes endocrine therapy (tamoxifen, aromatase inhibitors, etc) and CDK4/6 inhibitors. CDK4/6 inhibitors significantly prolong survival and is now the general accepted choice of drug in first line. Despite the enormous clinical benefit, there remains a substantial portion of patients who ultimately develop resistance against CDK4/6 inhibitors. Many mechanisms have been proposed for CDK4/6 inhibitor resistant, including upregulation of CDK4, CDK6, Cyclin E, and others. Recently studies have linked the hippo pathway to CDK6 upregulation and resulting in CDK4/6 inhibitor resistance. We have set up a kinase library screen for kinase inhibitors that downregulate CDK6 amplification and further assess for reversal of resistance. We discovered staurosporine and the PI3K inhibitor PIK-75 as small molecules that potently downregulate CDK6 expression in breast cancer cells. We further confirmed that these compounds reverse the resistance towards CDK4/6 inhibitors in CDK6 amplification cells. By kinase profiling combined with a small hairpin RNA (shRNA) screen, we discovered the mechanisms that resulted in CDK6 downregulation and modulation of the hippo pathway to reverse the CDK4/6 inhibitor resistance. Our discovery has implications for therapeutic development in this group of patients posing a huge unmet need.

Citation Format: Chung-Jen Yu, Yong-Ji Zhuang, Ting-Yi Lin, Ta-Chung Chao, Chun-Yu Liu, Ling-Ming Tseng, Jiun-I Lai. Discovery of mechanisms that modulate the hippo pathway and CDK6 expression in CDK4/6 inhibitor resistant breast cancer cells [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 5975.
Abstract 5977: Evaluation of the potential combination regimens for q901, a clinical stage selective cdk7 inhibitor, as a DNA damage repair inhibitor
Authors: Yu Donghoon , Jeon Yeejin , Lee YoonJi , Lee Seung-Joo , Kim Jaeseung , Jung Hyerim , Kim Tae-Kyung , Nam Kiyean
Published: April 2023

Background; Q901 is an extremely selective CDK7 inhibitor with potent single agent activity in the multiple in vivo tumor models including small cell lung cancer, cholangiocarcinoma, colon cancer, pancreatic cancer, hormone receptor positive breast cancer, castrate-resistant prostate cancer, and ovarian cancer. Q901 is currently in Phase 1/2 clinical trial for patients with selected advanced solid tumors (ClinicalTrials.gov: NCT05394103). In the preclinical models, Q901 treatment induces DNA damage response (DDR) impairment markers, particularly ones involved in the double-strand break repair mechanisms. This opens up broad range of possibilities for new combination regimen that could benefit from the accumulated DNA damages caused by Q901 treatment.

Methods: Various combinations have been tested to evaluate potential clinical regimens in multiple solid tumor models. Two different BRCA wild-type ovarian cancer A2780 and OVCAR3 models, an estrogen receptor positive MCF7 breast cancer model, or a syngeneic RENCA mouse renal cancer model was tested with Q901 in combination with poly ADP-ribose polymerase (PARP) inhibitor, selective estrogen receptor degrader (SERD), an estrogen receptor (ER) PROTAC degrader, or an anti-PD-1 antibody, respectively.

Results: In the A2780 model, the PARPi alone group showed 15% TGI (tumor growth inhibition), but significant tumor regression (104% TGI) was observed in the Q901 and PARPi combination. In the OVCAR3 model, the Q901 and PARPi combination group showed 105% TGI, whereas the PARPi alone group showed 38% TGI. In the MCF7 model, SERD alone showed 64% TGI and Q901 combination with SERD showed 94% TGI. Remarkable tumor regression (111% TGI) was observed in the MCF7 model combining ER PROTAC and Q901, whereas ER PROTAC alone group showed 55% TGI. In the RENCA model, the anti-PD-1 antibody alone group showed 13.2% TGI and the combination of Q901 and anti-PD-1 antibody showed 66% TGI. In summary, these preclinical model studies demonstrated significant additive/synergistic effects of the Q901 combination on tumor growth inhibition.

Conclusion: The non-clinical data demonstrate that Q901 not only has a potential as a single agent,but can also be used in combination with other therapeutic strategies to develop next-generation cancer therapeutics to potentially address unmet medical needs.

Citation Format: Donghoon Yu, Yeejin Jeon, YoonJi Lee, Seung-Joo Lee, Jaeseung Kim, Hyerim Jung, Tae-Kyung Kim, Kiyean Nam. Evaluation of the potential combination regimens for q901, a clinical stage selective cdk7 inhibitor, as a DNA damage repair inhibitor [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 5977.
Abstract 6002: Characterizing dynamic landscape of DNA methylation in ovarian high-grade serous carcinoma
Authors: Lahtinen Alexandra , Marchi Giovanni , Afenteva Daria , Holmström Susanna , Huhtinen Kaisa , Virtanen Anni , Hynninen Johanna , Häkkinen Antti , Hautaniemi Sampsa
Published: April 2023

Dysregulated DNA methylation (DNAm) is a hallmark of all cancer types, though the precise role of these aberrations in tumor progression and treatment response remains elusive. Advances in whole-genome bisulfite sequencing (WGBS) have enabled profiling methylomes at full resolution and stimulated interest to explore dynamics of DNAm across tumor sites and cancer treatment. A major challenge in the analysis of patient data is that a surgically removed tumor sample represents a mixture of cancer cells and microenvironment, and that the insights on DNAm changes are intertwined due to mixed effects from patient epigenomic background, tissue context, and treatment pressure. By using a computational approach, these effects can be decomposed into patient background, tissue site, and treatment phase specific changes, and controlled for natural microenvironment variation. retion, and action, as well as in several oncogenic signaling cascades. The analysis of the tissue profiles indicated that the progression of HGSC from the tissues-of-origin to the metastatic deposits is accompanied by drastic changes in DNAm profiles. Thus, when comparing ovaries and peritoneum, intra-abdominal tissues mesentery and omentum featured prominent change of promoter DNAm: loss in 10% and gain in 20% of genes. Interestingly, the genes with aberrant DNAm included ABC transporters implicated in pumping drugs across the cell membrane, as well as the members of MAPK and PI3K signaling cascades. The joint analysis of both treatment and tissue contributions suggests that the overall modest effect of NACT is further reduced in metastases, mediated via DNAm changes in genes involved in multidrug resistance.

Citation Format: Alexandra Lahtinen, Giovanni Marchi, Daria Afenteva, Susanna Holmström, Kaisa Huhtinen, Anni Virtanen, Johanna Hynninen, Antti Häkkinen, Sampsa Hautaniemi. Characterizing dynamic landscape of DNA methylation in ovarian high-grade serous carcinoma [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 6002.
Abstract 6018: Sequencing of 5-hydroxymethylcytosine to single base resolution
Authors: Evanich Daniel J. , Ponnaluri V. K. Chaithanya , Panchapakesa Vaishnavi , Erijman Ariel , Campbell Matthew A. , Dai Nan , Langhorst Bradley W. , Vaisvila Romualdas , Williams Louise
Published: April 2023

DNA methylation is an epigenetic regulator of gene expression with important functions in development and diseases such as cancer. The modified cytosines, 5-methylcytosine (5mC) and 5-hydroxymethylcytosine (5hmC) are identified using Illumina sequencing. Libraries are generated using either NEBNext® EM-seq™, an enzyme-based workflow, or by using sodium bisulfite conversion. These methods, however, cannot differentiate between 5mC and 5hmC. Distinguishing these modifications is important as there is increased interest in 5hmCs role in regulating gene expression. Methods currently exist to enable discrimination of 5mC and 5hmC, for example, oxBS-seq and TAB-seq. These methods still rely upon sodium bisulfite conversion and have reduced data quality due to fragmentation and loss of DNA. Here we describe an enzymatic method that enables specific detection of 5hmC. 5hmC is detected using two enzymatic steps. Firstly, 5hmCs are glucosylated, which protects them from subsequent deamination by APOBEC. In contrast, cytosines and 5mCs are deaminated resulting in their conversion to uracil and thymine, respectively. During Illumina sequencing deaminated cytosine and 5mC are read as thymine and the glucosylated 5hmC are read as cytosine. Subtractive analysis of 5hmC data from EM-seq data, which detects both 5mC and 5hmC, permits the identification of individual 5mC and 5hmC sites. 5hmC data were generated for inputs of 0.1 ng to 200 ng DNA isolated from E14 mouse embryonic stem cells and human brain. The 5hmC libraries had similar characteristics to EM-seq libraries, including expected insert sizes due to intact DNA molecules, low duplication rates and minimal GC bias. T4147 phage DNA was used as an internal control, as all cytosines are 5-hydroxymethylated, with 98-99% of cytosines identified as 5hmC. 5mC and 5hmC levels were also profiled during E14 cell differentiation for 10 days. Interestingly, 5hmC levels decreased whereas 5mC levels increased particularly during the first five days of differentiation. LC-MS/MS quantification of this same DNA mirrored the changes observed by sequencing. The ability to discriminate between 5mC and 5hmC will provide key insights into the role of these cytosine modifications in development and disease.

Citation Format: Daniel J. Evanich, V. K. Chaithanya Ponnaluri, Vaishnavi Panchapakesa, Ariel Erijman, Matthew A. Campbell, Nan Dai, Bradley W. Langhorst, Romualdas Vaisvila, Louise Williams. Sequencing of 5-hydroxymethylcytosine to single base resolution [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 6018.