bluuebunny/crossref_metadata_2025 huggingface dataset:
Abstract 706: Advancing radiotherapy strategies for high-risk rhabdomyosarcoma: Unveiling BCL-xL inhibition as a potent sensitizer to radiotherapy
Authors: Se Whee Park Sammy , Struyf Nona , Erkers Tom , Lehmann Sören , Meister Michael T. , Drost Jarno , Holstege Frank C. , Herold Nikolas , Östling Päivi , Stenman Jakob , Kallioniemi Olli , Henter Jan-Inge , Seashore-Ludlow Brinton , Karlsson Kasper
Published: March 2024

Pediatric soft-tissue sarcomas are a group of solid tumors that account for about 7% of childhood cancer cases. Rhabdomyosarcoma (RMS), the most common subtype, further stratifies into PAX3 or PAX7 and FOXO1 fusion-negative (FN) or fusion-positive (FP) RMS. FP RMS is the higher risk subtype and is more aggressive, metastatic, and less responsive to treatment. Front-line treatments for RMS FP patients have had limited progress in the recent years and existing treatments such as radiotherapy lacks tumor cell selectivity, resulting in toxic side effects.

To address this, we conducted a high-throughput drug screen to identify radiosensitizers to enhance efficacy of RMS radiotherapy. Using three patient-derived FP RMS tumoroids, we screened a comprehensive drug library with and without radiation, comprising 528 drugs at varying stages of pre-clinical and clinical trials. A validation screen of the top 29 drugs was conducted on a larger cohort of samples including eight patient-derived FP RMS tumoroids and three non-cancer models. The screening revealed that drugs targeting the anti-apoptotic protein B-cell lymphoma-extra-large (BCL-xL) exhibited both potent radiosensitization and cytotoxicity selectively for FP RMS. Notably, the sensitivity to BCL-xL inhibitors correlated with elevated BCL2L1 expression and PAX3 fusion status. Additionally, drugs targeting polo-like kinase 1 (PLK1) and other cell cycle-related genes exhibited robust radioprotective effects across both RMS and non-cancer models.

Our findings propose the therapeutic potential of BCL-xL inhibition, either as a standalone treatment or in combination with radiotherapy. In parallel, we show the protective potential of PLK1 inhibition to mitigate radiation toxicity. In the long term, we aspire to integrate the identified radiosensitizing and radioprotecting compounds with existing treatment regiments for RMS, maximizing precision and efficacy in RMS radiotherapy.

Citation Format: Sammy Se Whee Park, Nona Struyf, Tom Erkers, Sören Lehmann, Michael T. Meister, Jarno Drost, Frank C. Holstege, Nikolas Herold, Päivi Östling, Jakob Stenman, Olli Kallioniemi, Jan-Inge Henter, Brinton Seashore-Ludlow, Kasper Karlsson. Advancing radiotherapy strategies for high-risk rhabdomyosarcoma: Unveiling BCL-xL inhibition as a potent sensitizer to radiotherapy [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2024; Part 1 (Regular Abstracts); 2024 Apr 5-10; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2024;84(6_Suppl):Abstract nr 706.
Abstract 7074: Perturbed RNA N6-methyladenosine methylome in SETD2 mutant clear cell renal carcinoma
Authors: Shaikh Shafiq , Wagner Ryan , Hlady Ryan , Robertson Keith D.
Published: March 2024

Clear cell renal carcinoma (ccRCC), which arises from the epithelial cells of the proximal tubule represents 70% of all renal cell carcinoma. Mutations that drive ccRCC in conjunction with near ubiquitous and early VHL loss include, PBRM1 (33 - 45%), BAP1 (15%) and SETD2 (13%). Loss of SETD2 leads to diminished occupancy of H3K36me3 in active gene bodies, which is progressively deregulated in primary RCC and metastasis, linking loss of H3K36me to tumor progression and metastasis. N6-Methyladenosine (m6A) is an important post-transcriptional regulator of RNA transcript stability and half-life. Preliminary findings have shown H3K36me3 (deposited by SETD2), targeting the core m6A methyltransferase machinery (METTL3, METTL14 and WTAP) to the mRNA by virtue of its interaction to METTL14 subunit. The goal of this study is to investigate if H3K36me3 depletion as observed in SETD2 mutant ccRCC, leads to a deregulated m6A methylome. We further investigate how deregulated methylome in key pathways leads to cancer initiation and progression in ccRCC. To accomplish our goals, we generated isogenic ccRCC cell lines with H3K36me3 depletion by knocking out SETD2. RNA-immunoprecipitation and sequencing for m6A (MeRIP) showed global deregulation of m6A methylome in SETD2KO isogenic ccRCC cell lines. We also show a number of pro-tumorigenic genes and pathways including cell invasion, inflammation and stemness being differentially methylated and expressed in SETD2KO ccRCC cell lines. Furthermore, we show SETD2KO cells being more sensitive to METTL3 inhibition suggesting the relevance of m6A hypermethylated transcripts in promoting pro-tumorigenic and growth promoting processes in SETD2KO ccRCC cell lines.

Citation Format: Shafiq Shaikh, Ryan Wagner, Ryan Hlady, Keith D. Robertson. Perturbed RNA N6-methyladenosine methylome in SETD2 mutant clear cell renal carcinoma [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2024; Part 1 (Regular Abstracts); 2024 Apr 5-10; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2024;84(6_Suppl):Abstract nr 7074.
Abstract 7100: Quantitative proteomics reveal the mechanism of TTFields therapy for glioblastoma
Authors: Mei Qi , Gao Yanmei , Li Rong , Wang Chenxu , Hu Guangyuan , Huang He
Published: March 2024

Glioblastoma (GBM) is a highly malignant brain tumor, and conventional treatments such as surgical resection and chemotherapy have limited effectiveness, with a high rate of recurrence and poor prognosis for patients. In 2011, the FDA approved a novel therapy for recurrent GBM known as Tumor Treating Fields (TTFields). TTFields use alternating intermediate-frequency (200 kHz) and low-intensity (1-3 V/cm) electric fields to interfere with tumor cells, inducing apoptosis and inhibiting tumor growth. This study aimed to explore the molecular mechanisms of TTFields therapy at the systems biology level using quantitative proteomics technology. We analyzed U87 cells before and after TTFields treatment using quantitative proteomics and identified a total of 7679 proteins. Among them, 804 proteins were significantly upregulated after TTFields treatment, and 1141 proteins were significantly downregulated. Pathway enrichment analysis of differentially expressed proteins showed that upregulated proteins were mainly associated with cell proliferation, cell cycle, and transcription, possibly representing compensatory responses to the TTFields treatment. Further analysis revealed a significant upregulation of the tumor proliferation key protein PARP1, which was validated by protein immunoblotting experiments. Based on this analysis, we hypothesized that combining TTFields with Olaparib (a PARP1 inhibitor) might enhance the sensitivity of TTFields. Subsequently, we performed cell proliferation experiments to evaluate the concomitant application strategy, and the results demonstrated that Olaparib in combination with TTFields treatment significantly reduced the survival rate of U87 cells compared to single-drug treatment. In conclusion, using quantitative proteomics, this study revealed the biological processes underlying the inhibitory effects of TTFields on GBM proliferation. Under TTFields intervention, the expression of PARP1 was upregulated, and the use of a PARP1 inhibitor in combination significantly increased the sensitivity of TTFields. Therefore, this study not only elucidated the molecular mechanisms of TTFields therapy for GBM treatment at the systems biology level, but also provided support for further enhancing the efficacy of TTFields.

Citation Format: Qi Mei, Yanmei Gao, Rong Li, Chenxu Wang, Guangyuan Hu, He Huang. Quantitative proteomics reveal the mechanism of TTFields therapy for glioblastoma [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2024; Part 1 (Regular Abstracts); 2024 Apr 5-10; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2024;84(6_Suppl):Abstract nr 7100.
Abstract 7106: Distinct functions of EHMT1 and EHMT2 in cancer chemotherapy and immunotherapy
Authors: Kang Zhihua , Fu Pan , Ma Hui , Li Tao , Lu Kevin , Liu Juan , Ginjala Vasudeva , Romanienko Peter , Feng Zhaohui , Guan Ming , Ganesan Shridar , Xia Bing
Published: March 2024

EHTM1 (GLP) and EHMT2 (G9a) are closely related protein lysine methyltransferases often thought to function together as a heterodimer to methylate histone H3 and non-histone substrates in diverse cellular processes including transcriptional regulation, genome methylation, and DNA repair. Here we show that EHMT1/2 inhibitors cause DNA replication stress, accumulation of single-stranded replication gaps, increased expression of STING, emergence of cytosolic DNA, and activation of the cGAS-STING pathway. Remarkably, EHMT1/2 inhibition strongly potentiates the efficacy of alkylating chemotherapy and anti-PD-1 immunotherapy in mouse models of tripe negative breast cancer. Mechanistically, the effects of EHMT inhibition on DNA replication and alkylating agent sensitivity are largely caused by the loss of EHMT1-mediated methylation of DNA ligase I (LIG1), whereas the elevated STING expression is, at least part, caused by its promoter demethylation. Moreover, depletion of EHMT2, and not EHMT1, leads to STING induction, which is correlated with more complete demethylation of the STING1 promoter. On the other hand, depletion of EHMT1, and not EHMT2, results in cytosolic DNA accumulation, which presumably activates the cGAS-STING pathway. These results reveal distinct functions of the two EHMT paralogs and provide enlightening paradigms and corresponding molecular basis for combination therapies involving alkylating agents and immune checkpoint inhibitors.

Citation Format: Zhihua Kang, Pan Fu, Hui Ma, Tao Li, Kevin Lu, Juan Liu, Vasudeva Ginjala, Peter Romanienko, Zhaohui Feng, Ming Guan, Shridar Ganesan, Bing Xia. Distinct functions of EHMT1 and EHMT2 in cancer chemotherapy and immunotherapy [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2024; Part 1 (Regular Abstracts); 2024 Apr 5-10; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2024;84(6_Suppl):Abstract nr 7106.
Abstract 7109: Fasn inhibition enhances the efficacy of chemotherapy by inhibiting DNA damage response in colorectal cancer
Authors: Banerjee Moumita , Zaytseva Yekaterina , Hasani Sumati , Ryachahou Piotr , Izumi Tadahide , Evers B. Mark , Gao Tianyan
Published: March 2024

Altered lipid metabolism plays a significant role in progression of colorectal cancer, however, its role in chemotherapy resistance remains poorly understood. We previously identified FASN, the rate limiting enzyme in de novo lipogenesis, as an important regulator of colorectal cancer progression. However, TVB-3664, a potent and specific small molecule inhibitor of FASN, showed limited effect in reducing tumor burden in pre-clinical studies as a stand-alone therapy. Therefore, the goal of this study is to evaluate the combinatorial efficacy of TVB-3664 in improving the therapeutic indices of standard chemotherapy in colorectal cancer. Using a combination of colon cancer cell lines and patient derived organoids we showed that TVB-3664 potentiates chemotherapy drug-induced double-strand DNA breaks (DSBs) and apoptotic cell death by altering histone acetylation levels. In addition, TVB-3664 treatment blocked DNA damage response (DDR) by decreasing ATM expression and Chk1/2 phosphorylation. Mechanistically, inhibition of FASN prevents the activation of both DSB repair pathways, namely homologous recombination (HR) and classical non-homologous end joining (NHEJ), by blocking BRCA1 and 53BP1 recruitment to γH2AX foci, respectively. Moreover, we found that the inhibitory effect of TVB-3664 on DNA repair requires ACLY-dependent acetylation as genetic or pharmacological inhibition of ACLY attenuates TVB-3664-inducded DNA damage. Functionally, combining TVB-3664 with irinotecan synergistically decreased xenograft tumor growth and significantly delayed tumor relapse after withdrawal of therapy. Taken together, we identified a novel mechanism by which FASN inhibitors improve the efficacy of conventional chemotherapy by blocking DNA damage response to delay tumor recurrence and overcome a major challenge in treating colorectal cancer patients.

Citation Format: Moumita Banerjee, Yekaterina Zaytseva, Sumati Hasani, Piotr Ryachahou, Tadahide Izumi, B. Mark Evers, Tianyan Gao. Fasn inhibition enhances the efficacy of chemotherapy by inhibiting DNA damage response in colorectal cancer [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2024; Part 1 (Regular Abstracts); 2024 Apr 5-10; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2024;84(6_Suppl):Abstract nr 7109.
Abstract 7120: Investigating PARP1 selective inhibitor DSB1559 efficacy in BRCA1-associated triple negative breast cancer
Authors: Nelson Adam , Zheng Kelly F. , Wanderley Carlos , Michaud Daniel E. , Cowley Phillip M. , Campbell Gillian M. , McGuinness Barry E. , Wise Alan , Guerriero Jennifer
Published: March 2024

Background: Poly (ADP-ribose) polymerase inhibitors (PARPi) have improved outcomes of BRCA-associated breast cancer. However, toxicity can limit patient response as well as the ability to be combined with other therapeutics. Current PARPi broadly inhibit members of the PARP protein family, however only inhibition of PARP1 is needed for PARPi-mediated cell death. As inhibition of PARP2 is associated with hematological toxicity seen in PARPi patients, selectively inhibiting PARP1 may reduce PARPi toxicity and lead to better outcomes for patients. Here we characterize and test the efficacy of the PARP1 selective inhibitor (PARP1i) DSB1559 compared to Olaparib in BRCA1-associated triple negative breast cancer (TNBC).

Methods: NanoBRET, PARylation, and pharmacokinetic/pharmacodynamic assays were performed to examine DSB1559 potency, PARP family selectivity, and in vivo behavior. Immunocompetent FVB/n mice bearing BRCA1-deficient TNBC (CMV-Cre;Brca1f/f;Trp53f/f) tumors were treated daily with PARPi (Olaparib) or PARP1i (DSB1559) for 60 days and followed for survival. CellTiter-Glo and qPCR assays were used to examine DSB1559 effect on cell viability and STING activation, respectively.

Results: DSB1559 is a novel and potent PARP1 inhibitor which has superior selectivity for PARP1 compared to Olaparib and PARP1i AZD5305. DSB1559 has highly desirable physico-chemical and in vitro ADME properties, which translate to high oral bioavailability and moderate clearance in rodent PK studies. Additionally, DSB1559 demonstrated superior tumor residence time and tumor/plasma ratio compared to AZD5305. Compared to Olaparib, DSB1559 significantly increased STING activation and reduced cell viability in BRCA1-associated murine TNBC cells. BRCA1-associated TNBC-bearing immune-competent mice treated with DSB1559 had significantly reduced tumor growth and increased overall survival (median survival 74 days) compared to Olaparib (median survival 18 days). Analysis of changes in the tumor microenvironment are ongoing.

Conclusions: Taken together, DSB1559 is a highly selective PARP1 inhibitor that demonstrates superior efficacy compared to Olaparib in a BRCA1-associated immune-competent TNBC model.

Citation Format: Adam Nelson, Kelly F. Zheng, Carlos Wanderley, Daniel E. Michaud, Phillip M. Cowley, Gillian M. Campbell, Barry E. McGuinness, Alan Wise, Jennifer Guerriero. Investigating PARP1 selective inhibitor DSB1559 efficacy in BRCA1-associated triple negative breast cancer [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2024; Part 1 (Regular Abstracts); 2024 Apr 5-10; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2024;84(6_Suppl):Abstract nr 7120.
Abstract 7133: Phenomics-enabled discovery and optimization of small-molecule RBM39 degraders as an alternative to CDK12 targeting in high-grade serous ovarian cancer (HGSOC)
Authors: Neumann Chase , Shankaran Harish , Nadella Kiran , Biette Kelly , Rowley Shane , Gardner Ethan , Swaidani Shadi , Manda Vamshi , Chen Lu , Beshnova Daria , Saeed Ashraf , Bailey Christopher , Paulsen Janet , Rearden Paul , Brooks Carl , Bhandari Ashish , Gibson Chris , Schaevitz Laura , Haque Imran , Donnella Hayley , Cuccarese Michael , Evangelista Marie
Published: March 2024

Phenomics-enabled drug discovery is a powerful approach to identify novel targets and relationships previously unappreciated in biology. We built a phenomics platform leveraging high content microscopy and generated unique phenoprints for >7,000 genes using CRISPR/Cas9 technology and a diverse chemical library of >1 million compounds. This phenomics dataset contains an unprecedented quantity of gene-gene, gene-compound, and compound-compound relationships. We applied our phenomics platform and an inference based target-agnostic drug discovery approach to discover novel genes and small molecules that mimic CDK12 inhibition, an important transcriptional regulator of DNA damage response, while avoiding inhibition of the paralog CDK13. We discovered a novel and unappreciated association between CDK12 and RBM39 as well as small molecule degraders of RBM39. We established structure activity relationships (SAR) solely leveraging our phenomics platform, to generate REC-1170204, an RBM39 degrader. Unlike inhibitors of CDK12, we show that REC-1170204 does not directly inhibit CDK12, CDK13 or other kinases. However, REC-1170204 demonstrates candidate quality properties with improved potency, selectivity, and drug-like characteristics. Finally we show that REC-1170204 shows efficacy in High Grade Serous Ovarian cancer (HGSOC) pre-clinical models and synergizes with PARP inhibition in a PARP-resistant patient-derived xenograft (PDX) model. Together, our data suggests that targeting RBM39 may provide an attractive and safer approach to targeting CDK12 in the clinic.

Citation Format: Chase Neumann, Harish Shankaran, Kiran Nadella, Kelly Biette, Shane Rowley, Ethan Gardner, Shadi Swaidani, Vamshi Manda, Lu Chen, Daria Beshnova, Ashraf Saeed, Christopher Bailey, Janet Paulsen, Paul Rearden, Carl Brooks, Ashish Bhandari, Chris Gibson, Laura Schaevitz, Imran Haque, Hayley Donnella, Michael Cuccarese, Marie Evangelista. Phenomics-enabled discovery and optimization of small-molecule RBM39 degraders as an alternative to CDK12 targeting in high-grade serous ovarian cancer (HGSOC) [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2024; Part 1 (Regular Abstracts); 2024 Apr 5-10; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2024;84(6_Suppl):Abstract nr 7133.
Abstract 7142: Optimizing estradiol dimers through linker design: Enhancing anticancer efficacy by targeting microtubule dynamics
Authors: Dzubak Petr , Jurášek Michal , Řehulka Jiří , Gurská Soňa , Polishchuk Pavel , Dráber Pavel , Huml Lukáš , Drašar Pavel B. , Ivanova Alexandra , Mokshyna Olena , Hrubá Lenka , Hajdúch Marián
Published: March 2024

We investigated the development and anticancer efficacy of novel estradiol dimers (ED), targeting the dynamics of microtubules—an essential factor in effective cancer therapy. Employing copper-catalyzed azide-alkyne cycloaddition (CuAAC), two distinct series of ED variants were synthesized. The initial series included thirteen dimers, derived from 17α-ethinyl estradiol, incorporating varied five-atom linkers with central carbon, nitrogen, or oxygen atoms, and, in some instances, modified with benzyl groups containing hydroxy or methoxy substituents. These dimers were evaluated for their cytotoxic properties and impact on the cell cycle, revealing certain structures with substantial efficacy, compared with the original ED. The subsequent series, encompassing twelve variants, concentrated on structural alterations within the aromatic bridges linking the estradiol units. In this series, linkers of simpler composition and fewer substituents demonstrated increased selectivity and potency against cancer cell lines, particularly ED3 and ED5, which exhibited cytotoxicity levels comparable to established tubulin inhibitors. Both dimers series showed the ability to disrupt microtubule assembly and mitotic processes, confirmed through cell-based evaluations and in vitro tubulin assembly assays. Complementing these results, in silico modelling and binding free energy calculations exhibited a strong correlation with the demonstrated biological activities, especially in terms of binding affinity to the colchicine site. Collectively, these findings highlight the promise of estradiol dimers as potential anticancer agents and emphasize the critical role of linker structures in enhancing their biological effectiveness. Work was supported by the Ministry of Education, Youth and Sports of the Czech Republic through infrastructural projects (CZ-OPENSCREEN - LM2018130; EATRIS-CZ - LM2018133) and National Institute for Cancer Research (Programme EXCELES, ID Project No. LX22NPO5102) - Funded by the European Union - Next Generation EU.

Citation Format: Petr Dzubak, Michal Jurášek, Jiří Řehulka, Soňa Gurská, Pavel Polishchuk, Pavel Dráber, Lukáš Huml, Pavel B. Drašar, Alexandra Ivanova, Olena Mokshyna, Lenka Hrubá, Marián Hajdúch. Optimizing estradiol dimers through linker design: Enhancing anticancer efficacy by targeting microtubule dynamics [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2024; Part 1 (Regular Abstracts); 2024 Apr 5-10; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2024;84(6_Suppl):Abstract nr 7142.
Abstract 7144: BR0L075 (SCB01A), a microtubule inhibitor targeting the colchicine binding site, induces immunogenic cell death
Authors: Bano Ayesha , Liu Xinli
Published: March 2024

BPR0L075 (also known as SCB01A in phase II clinical trial) is a novel anti-microtubule drug with potent anti-tumor and anti-angiogenic activity in vitro and in vivo. BPR0L075 inhibits tubulin polymerization (microtubule de-stabilizer) by binding to the colchicine-binding site of tubulin. We previously reported that BPR0L075 overcomes paclitaxel (microtubule stabilizer) resistance and induces vascular-disruption in vitro and in vivo. We observed that breast and ovarian cancer cells treated with BPR0L075 in vitro induced key hallmarks of immunogenic cell death (ICD). In both human and murine, ovarian (OVCAR3, ID8) and breast cancer (MDA-MB-231, 4T1) cell lines, treatment with low nanomolar concentration of BPR0L075 significantly decreased the viability of cells evidenced by SRB assay. Using a known ICD inducer mitoxantrone as a positive control, both flow cytometric analysis and immunofluorescence staining assays demonstrated that BPR0L075 treatment led to a time- and dose-dependent translocation of calreticulin, an endoplasmic reticulum chaperone protein, on the plasma membrane of dying cells which sends an “eat me” signal. The BPR0L75 treatment also led to secretion of High Mobility Group Box 1 (HMGB1), a non-histone chromatin-binding nuclear protein, as well as adenosine triphosphate (ATP) from the cell into the extracellular space using ELISA assay. These damage-associated molecular patterns (DAMPs) are known to lead to antigen-presenting cell recruitment, phagocytosis, maturation of dendritic cells, and chemotaxis. The appearance of the ICD biomarkers (calreticulin exposure, HMGB1 and ATP release) supports BPR0L075 as an ICD-inducing antimitotic agent. Further in vivo studies are warranted to delineate the effect of BPR0L075 on immune activation and recruitment of immune cells to the tumor microenvironment.

Citation Format: Ayesha Bano, Xinli Liu. BR0L075 (SCB01A), a microtubule inhibitor targeting the colchicine binding site, induces immunogenic cell death [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2024; Part 1 (Regular Abstracts); 2024 Apr 5-10; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2024;84(6_Suppl):Abstract nr 7144.
Abstract 721: Novel and distinctive anti-HLA-G antibody, IMB-201, inhibits tumor progression by breaking immune escape mechanism and enhancing cytotoxicity
Authors: Kang Hyeonju , Baek Seungil , Lee Inyoung , Ju Manseok , Han Seonggu , Han Eunjung , Kang Sungmuk , Koo Jihye , Kim Yoojin , Park Suho , Cho Eunyoung , Lee Chungmin , Ha Gyongsik
Published: March 2024

Human leukocyte antigen-G (HLA-G), a type of MHC class I, is known to be expressed only in the placenta and acts as an immune suppressor to protect the fetus from maternal immunity. Recent studies have revealed that HLA-G suppresses various tumor-infiltrated immune cells, such as T cells, NK cells, and macrophages, through interaction with inhibitory receptors Ig-like transcript 2 (ILT2). HLA-G has recently emerged as an attractive anticancer target because HLA-G is a highly specific tumor antigen and has a role in immune evasion of cancer cells. Additionally, the expression pattern of ILT2 differs from that of PD-1 in tumor-infiltrated CD8+ T cells. Therefore, an HLA-G targeting strategy is expected to be an alternative to address unmet medical needs for those not responding to anti-PD(L)1 therapies. In this study, we screened the novel anti-HLA-G antibody, IMB-201, using phage display technique and confirmed that IMB-201 selectively bound to the membrane HLA-G of cancer cells. It exhibited superior activities in ILT2 blockade, NK cell activation, and antibody-dependent cellular cytotoxicity (ADCC) compared to the reference antibody. Furthermore, IMB-201 demonstrated more efficient suppression of tumor growth compared to the reference antibody in subcutaneous gastric cancer xenograft model. Notably, the group administered IMB-201 with NK92MI cells (CD16-negative NK cells) in the ovarian cancer xenograft model showed stronger tumor growth inhibition compared to the group treated only IMB-201, indicating that NK92MI cells were activated by IMB-201. This result indicates that one of the mechanisms of action (MoA) of IMB-201 worked effectively in vivo. In addition, to maximize the efficacy of IMB-201 and differentiate it from competitors, various efficacy enhancing modality could be applicable. We aim to develop biopharmaceuticals by selecting the optimized modality. Based on these results, IMB-201 could overcome the limitations of existing cancer immunotherapy, potentially becoming highly influential in cancer treatment as a novel immune checkpoint inhibitor. It is also expected to exhibit higher anticancer efficacy in combination therapy with immune checkpoint inhibitors.

Citation Format: Hyeonju Kang, Seungil Baek, Inyoung Lee, Manseok Ju, Seonggu Han, Eunjung Han, Sungmuk Kang, Jihye Koo, Yoojin Kim, Suho Park, Eunyoung Cho, Chungmin Lee, Gyongsik Ha. Novel and distinctive anti-HLA-G antibody, IMB-201, inhibits tumor progression by breaking immune escape mechanism and enhancing cytotoxicity [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2024; Part 1 (Regular Abstracts); 2024 Apr 5-10; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2024;84(6_Suppl):Abstract nr 721.
Abstract 7217: Orthogonal targeting of NAD metabolism and ErbB2/ErbB3 signaling in pancreatic adenocarcinoma
Authors: Chougoni Kranthi Kumar , Kim Nari , Dcona Martin M. , Dcona Diana T. , Malhotra Gautam K. , Grossman Steven R.
Published: March 2024

Background: Pancreatic ductal adenocarcinoma (PDAC) is now the third leading cause for cancer related deaths in the US with a five-year survival of ~10% that has improved little over the past 20 years. Thus, there is a critical need to fully understand mechanisms of PDAC tumorigenesis in order to develop novel and effective therapies. Nicotinamide Adenine Dinucleotide (NAD), a critical co-factor and energy source required for cellular processes such as DNA repair and epigenetic regulation is frequently upregulated in PDAC. We recently reported a novel oncogenic axis connecting the NAD-dependent epigenetic regulator and oncogene C-terminal Binding Protein (CtBP) with the EGFR family growth factor receptors ErbB2/ErbB3 (ErbB2/3) in PDAC cells and tumors. Indeed, previous reports have shown ~30% of human PDAC tumors overexpress ErbB3, and our work has shown that PDAC tumor cell growth relies on maintenance of ErbB3 expression. Moreover, we show that the majority of PDAC cells with high levels of ErbB3 expression are uniquely sensitive to the ErbB2-targeted tyrosine kinase inhibitor lapatinib. In addition, we have recently shown that co-targeting NAD and CtBP with small molecule inhibitors is an effective and synergistic therapeutic strategy in PDAC model systems.

Methods and Results: In the current study, we demonstrate NAD-dependent regulation of ErbB2/3 signaling in PDAC cells in vitro. Upon depletion of NAD using the NAD salvage pathway inhibitor GMX1778 in human PDAC cells, we observed a striking coordinate decrease in the levels of both ErbB2 and ErbB3. Furthermore, stable knockdown of the limiting enzyme of the complementing Preiss-Handler NAD synthetic pathway, Nicotinamide Adenine Phosphoribosyl Transferase, also resulted in robust downregulation of ErbB2/3 protein levels, suggesting a direct linkage in PDAC cells between the status of NAD metabolism and synthesis and activity of the key growth factor receptors ErbB2/3.

Conclusions: Our findings highlight the novel connection between NAD and ErbB2/3 in PDAC. Furthermore, our data suggests combining chemical or biologic inhibitors of ErbB2/3 with an inhibitor of NAD synthesis may result in highly synergistic cytotoxic effects in PDAC cells and PDAC preclinical models. Such data could support the future development of a human PDAC clinical trial repurposing agents targeting NAD synthesis and ErbB2/3 that are already FDA-approved or currently in clinical testing.

Citation Format: Kranthi Kumar Chougoni, Nari Kim, Martin M. Dcona, Diana T. Dcona, Gautam K. Malhotra, Steven R. Grossman. Orthogonal targeting of NAD metabolism and ErbB2/ErbB3 signaling in pancreatic adenocarcinoma [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2024; Part 1 (Regular Abstracts); 2024 Apr 5-10; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2024;84(6_Suppl):Abstract nr 7217.
Abstract 7220: Emerging combination strategy: FANCI suppression induces PARP1 redistribution to enhance efficacy of PARP inhibitors in breast cancer
Authors: Huang Yu-zhou , Sang Ming-Yi , Xi Pei-Wen , Xu Ruo-Xi , Cai Meng-Yuan , Wang Zi-Wen , Zhao Jian-Yi , Li Yi-Han , Wei Ji-Fu , Ding Qiang
Published: March 2024

While polyADP-ribose polymerase (PARP) inhibitors have made advancements in the treatment of breast cancer, challenges such as chemotherapy resistance and limited application persist. Fanconi AnemiaComplementation Group I (FANCI), a DNA repair protein associated with breast cancer development, represents a potential target for novel combination therapeutic strategies. However, the role of FANCI in breast cancer and its impact on the efficacy of PARP inhibitors require further investigation.

In our study, we analyzed FANCI expression in breast cancer tissues and cell lines, and its correlation with clinical parameters and patient prognosis. Lentiviral vectors were utilized and functional assays were performed to evaluate the effects of FANCI modulation on breast cancer cell growth and migration. Co-immunoprecipitation assays and protein interaction analysis were conducted to identify the interaction between FANCI and PARP1 and determine the specific binding region. The functionality and nuclear distribution of PARP1 were assessed upon FANCI modulation. Finally, the sensitivity of breast cancer cells to the PARP inhibitor talazoparib upon FANCI knockdown was evaluated in vitro and in vivo.

Our findings demonstrated that FANCI was overexpressed in breast cancer and associated with poor prognosis. FANCI significantly promoted breast cancer cell proliferation both in vitro and in vivo. We identified the interaction between FANCI and PARP1, specifically at the FANCI helical domain 2 binding site. FANCI knockdown led to reduced nuclear localization of PARP1 and decreased PARP1 activity. Importantly, combination treatment with FANCI knockdown and talazoparib significantly inhibited cancer growth in vitro and in vivo. Additionally, we found that the cyclin-dependent kinase (CDK) 4/6 inhibitor palbociclib, which effectively suppresses FANCI protein expression, exhibited a robust synergistic effect with talazoparib both in vitro and in vivo.

In conclusion, FANCI is a novel therapeutic target for breast cancer. Suppression of FANCI regulates PARP1 redistribution and activity, making breast cancer cells more responsive to PARP inhibitors. This combination therapeutic strategy shows potential in enhancing the effectiveness of PARP inhibitors for breast cancer treatment, regardless of BRCA mutations.

Citation Format: Yu-zhou Huang, Ming-Yi Sang, Pei-Wen Xi, Ruo-Xi Xu, Meng-Yuan Cai, Zi-Wen Wang, Jian-Yi Zhao, Yi-Han Li, Ji-Fu Wei, Qiang Ding. Emerging combination strategy: FANCI suppression induces PARP1 redistribution to enhance efficacy of PARP inhibitors in breast cancer [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2024; Part 1 (Regular Abstracts); 2024 Apr 5-10; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2024;84(6_Suppl):Abstract nr 7220.
Abstract 7229: In situ trans-differentiation therapy effectively terminates the growth and recurrence of glioblastoma
Authors: Zhao Huihui , Li Zhaozhong , Zhang Xiaohui , Zhao Congjian , Zhu Zhao , Liu Yueguang
Published: March 2024

Background: Glioblastoma (GBM) is the most common and malignant brain tumor in adults, and remains almost invariably lethal due to its aggressive and invasive nature. The standard first-line treatment for GBM is surgical resection, postoperative radiotherapy and chemotherapy with temozolomide (TMZ), but it exhibits short-term effects due to a high recurrence rate of GBM is its (up to almost one hundred percent). This long-existing clinical challenge has demanded development of novel and effective therapeutic strategies. In situ cell trans-differentiation therapy which direct reprograms tumor cells into terminally differentiated benign cells represents a novel therapeutic strategy for tumors. We further explored the application of in situ trans-differentiation therapy for glioblastoma.

Results: In the present study, we have conducted a large-scale screen for transcription factors which can reprogram glioblastoma cells into terminally differentiated postmitotic cells. Over-expression of a combo of selected trans-differentiation factors that possess the highest trans-differentiation rate effectively reprogramed human glioma cells into neuronal-like cells with characteristic neuronal morphology and expression of neuronal genes in culture, as a result substantially reducing glioblastoma cell proliferation and tumor development after orthotopic transplantation. With replicable recombinant oncolytic virus vectors, product candidate can specifically replicate in glioma cells, therefore transcription factors can be specifically delivered to all tumor cells rather than normal cells. In either cell line-derived or patient-derived tumor tissue xenograft models, intra-tumoral injection product candidate inhibited glioblastoma cells growth at an exceptionally high efficiency. Remarkably product candidate combination with TMZ resulted in a complete regression of tumor and no recurrence was observed in animal model cases.

Conclusion: In summary, our findings have established a novel therapeutic approach for treating glioblastoma by terminating sustained proliferation of glioma cells via transcription factors-induced in situ cell trans-differentiation and supported further clinic investigations in patients.

Citation Format: Huihui Zhao, Zhaozhong Li, Xiaohui Zhang, Congjian Zhao, Zhao Zhu, Yueguang Liu. In situ trans-differentiation therapy effectively terminates the growth and recurrence of glioblastoma [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2024; Part 1 (Regular Abstracts); 2024 Apr 5-10; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2024;84(6_Suppl):Abstract nr 7229.
Abstract 7309: Edible mushroom derived antioxidant, vialinin A, prevents melanoma growth and invasion
Authors: Vo Alexandria , Zafar Amber
Published: March 2024

Melanoma is the second leading invasive cancer with a greater capacity to invade and metastasize. Increased oxidative stress has been shown to be involved in melanoma growth and spread. Several antioxidants, vitamins, and other phytochemicals have been shown to prevent melanoma cancer growth. However, the chemopreventive efficacy of vialinin-A, an edible mushroom-derived p-terphenyl antioxidant, on cancer growth and invasion is unknown. We, therefore, hypothesized that with its potent anti-oxidative and anti-inflammatory actions, vialinin-A could prevent melanoma cell growth and invasion. B16-F10 melanoma cells were treated without or with various concentrations of vialinin-A. The cell viability was determined by MTT assay and apoptosis by annexin-V staining. Our results suggest that vialinin-A prevents melanoma cell growth in a dose-dependent manner. Vialinin-A also prevents the invasion and migration of melanoma cells. Further, vialinin-A prevents the generation of reactive oxygen species and promotes apoptosis of B16-F10 cells. In addition, vialinin-A increases the activation of caspase-3 and cleavage of PARP protein. Vialinin-A also regulated the expression of various anti-apoptotic, pro-apoptotic, and inflammatory markers in melanoma cells. In conclusion, our results suggest that vialinin-A, through regulating oxidative stress-modulated apoptotic pathways, could prevent melanoma cell growth and invasion and has potential chemopreventive properties.

Citation Format: Alexandria Vo, Amber Zafar. Edible mushroom derived antioxidant, vialinin A, prevents melanoma growth and invasion [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2024; Part 1 (Regular Abstracts); 2024 Apr 5-10; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2024;84(6_Suppl):Abstract nr 7309.
Abstract 7315: Collaboration between NIDCR and NCI promotes innovative research in head and neck cancers
Authors: Chen Zhong , Wang Wendy
Published: March 2024

The Oral and Salivary Cancer Biology Program at National Institute of Dental and Craniofacial Research (NIDCR) supports research activities that improve early detection and treatment of cancers of the oral cavity, pharynx, and salivary glands. We promote basic science research on the regulatory pathways and transformation process of pre-cancerous lesions to primary and metastatic tumors, as well as translational science activities to develop novel technologies and therapeutics for the diagnosis, prognosis, and treatment of cancers. NIDCR and National Cancer Institute (NCI) have established long-term collaborations to promote and co-manage awards funded through The Immuno-Oncology Translational Network (IOTN) of the Cancer Moonshot program, Specialized Programs of Research Excellence (SPOREs), and Early Detection Research Network (EDRN) program. NIDCR and NCI have co-developed Notice of Special Interest (NOSI) and Request for Application (RFA) on Advancing Head and Neck Cancer Early Detection Research (AHEAD), which co-funded seven research projects to discover and develop molecular signatures of early detection and diagnosis of HPV positive and negative head and neck cancer (HNC). To further strengthen NIDCR and NCI collaboration, we jointly organized the symposium to promote HNC early detection and biomarker identification at the American Head and Neck Society (AHNS) 11th International Conference on Head and Neck Cancer in July 2023. In addition, NIDCR has organized a symposium to promote HNC research at the 2024 annual meeting of American Association for Dental, Oral, and Craniofacial Research (AADOCR) to celebrate the 75th anniversary of NIDCR. This AADOCR symposium will feature the collaboration between NIDCR with NCI in the Cancer Moonshot program focusing on immunoprevention and immunotherapy, HNC early detection and biomarker research in AHEAD and EDRN, and investigation of HPV positive and negative HNC and clinical translational research under SPORE. At this 2024 AACR annual meeting, NIDCR and NCI program directors will present the collaborations and funding opportunities between NIDCR and NCI, focusing on early cancer detections and biomarker discovery and validation. Collaboration between NIDCR with NCI provides a unique opportunity to promote basic, translational, and clinical research in advancing HNC early detection to benefit cancer patients.

Citation Format: Zhong Chen, Wendy Wang. Collaboration between NIDCR and NCI promotes innovative research in head and neck cancers [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2024; Part 1 (Regular Abstracts); 2024 Apr 5-10; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2024;84(6_Suppl):Abstract nr 7315.
Abstract 737: Arrays of 3D ECM-embedded fibroblast clusters for characterization of ECM remodeling by primary cancer associated fibroblasts and for evaluation of drugs attenuating fibrotic ECM remodeling
Authors: Danen Erik H. J.
Published: March 2024

Chronic fibrosis can lead to organ failure and is also associated with cancer, where the tumor is interpreted as a chronic wound, and the fibrotic tumor environment drives disease progression. Fibrosis occurs in an inflammatory environment where macrophages are activated, and fibroblasts transdifferentiate into myofibroblasts. Activated macrophages stimulate the myofibroblasts to produce excessive amounts of collagen-rich ECM. Changes in proteolytic activity and high contractile forces that myofibroblasts apply onto their tissue environment, further lead to extensive remodeling of the healthy tissue matrix into a scar matrix. Similar activity is displayed by cancer-associated fibroblasts (CAFs) in tumors. Intervening with the excessive ECM remodeling by activated fibroblasts represents a candidate therapeutic strategy to normalize the architecture of fibrotic and malignant tissues. We developed a screening platform using an assay based on automated image guided injection of clusters of fibroblasts in wells of multi-well plates preloaded with a collagen-rich ECM network. The identical x-y-z position, spacing, and size of the ECM-embedded fibroblast clusters in each well facilitates automated real time confocal microscopy and quantitative image analysis algorithms. We use this setup for automated quantitative analysis of ECM remodeling activity of CAFs obtained from early-stage colorectal cancer patients. We find that CAFs from these early-stage lesions display increased ECM remodeling capacity as compared to patient matched normal fibroblasts. We then use the same setup to screen a series of compounds for their ability to attenuate ECM remodeling triggered by TGFb-activated primary human fibroblasts. We detect known and novel pathways, and we identify small molecule inhibitors with a promising efficacy versus toxicity profile. Lastly, we detect a previously unknown concentration-dependent side effect of ALK5 inhibitors that leads to the identification of a novel profibrotic pathway and indicates that caution is warranted for the clinical application of ALK5 inhibitors.

Citation Format: Erik H. J. Danen. Arrays of 3D ECM-embedded fibroblast clusters for characterization of ECM remodeling by primary cancer associated fibroblasts and for evaluation of drugs attenuating fibrotic ECM remodeling [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2024; Part 1 (Regular Abstracts); 2024 Apr 5-10; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2024;84(6_Suppl):Abstract nr 737.
Abstract 7381: Mathematical modeling and validation of mechanistic target of rapamycin and N-myristoyltransferase signaling pathways in breast cancer
Authors: Meher Abinash , Shrivastav Shailly Varma , Agarwal Anouska , Dube Sheen , Portet Stephanie , Shrivastav Anuraag
Published: March 2024

Hormone receptor-positive (HR+) breast cancer (BC) makes up approximately 65% of all breast cancers diagnosed. The prognosis for early-stage disease is excellent with mainstay endocrine therapy (ET). Despite the effectiveness of standard ET, as many as 41% of HR+ early-stage BC diagnosed women will experience distant recurrence. The resistance to endocrine therapy and recurrence is partly attributed to the activation of the insulin pathway and the independence of HR+ BC cells on the ER pathway for their growth. Earlier, we demonstrated the crosstalk between insulin/mTOR and ER pathways. N-myristoyltransferase (NMT) exists in humans in two paralogues (NMT1 and NMT2) that catalyze myristoylation reaction. Recent studies from our laboratory demonstrated that NMTs are downstream targets of insulin and ER pathways. In this study, we have designed mathematical models using differential equations to study the activation of the insulin pathway and its effect on NMT. The mathematical modeling incorporated the partition of cellular organelles and the sequential flow of information with cascades of equations representing signaling reactions. The activated mathematical model was designed by activating the insulin receptor (IR) or insulin-like growth factor receptor (IGF1R) and compared with the control model. The mathematical models were validated by wet lab experiments. The HR+ BC cells (MCF7) were treated with insulin or IGF1 for short term and long term. The expression patterns of NMT1, NMT2, and mTOR were determined by cell fractionation and western blot analysis. The results revealed the correlation between the differential expression patterns of NMTs and the proliferation of MCF7 cells when the insulin pathway was activated by insulin or IGF. The mathematical modeling was validated by simulations and data fittings. The results demonstrated that differential equation-based mathematical modeling could predict the NMT-related oncogenic changes in HR+ BC cells.

Citation Format: Abinash Meher, Shailly Varma Shrivastav, Anouska Agarwal, Sheen Dube, Stephanie Portet, Anuraag Shrivastav. Mathematical modeling and validation of mechanistic target of rapamycin and N-myristoyltransferase signaling pathways in breast cancer [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2024; Part 1 (Regular Abstracts); 2024 Apr 5-10; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2024;84(6_Suppl):Abstract nr 7381.