bluuebunny/crossref_metadata_2025 huggingface dataset:
Abstract 4789: Spin oscillating magnetic field sOMF of oncomagnetic device selectively kills glioma cancer cells by inducing oxidative stress and DNA damage
Authors: Hambarde Shashank , Pandey Arvind , Baskin David S. , Helekar Santosh A.
Published: April 2023

A new noninvasive therapeutic device developed in our laboratory called the Oncomagnetic device provides a novel approach to glioblastoma (GBM) treatment. It involves repeated stimulation with oscillating magnetic fields (sOMF) produced by spinning permanent magnets. It is technologically and mechanistically distinct from the Optune® device approved by the FDA for the treatment of newly diagnosed and recurrent GBM. Use of this device in end-stage GBM patients reversed the progression of their recurrent tumor causing >30% reduction in its contrast-enhanced volume within 4-6 weeks of treatment. Mice with implanted mouse glioma cells in their brains also showed marked reduction in tumor size, increased survival (p< 0.05, n = 10) and higher DNA damage (γ-H2AX foci) after sOMF treatment with a whole-body stimulation method developed by us. Normal mice exposed to sOMF for 4 months had no adverse effects on the brain and other organs. In-vitro, sOMF markedly increased reactive oxygen species (ROS) levels in cancer cells leading to the selective death of these cells, while sparing normal neurons and astrocytes. Detection of γ-H2AX and 53BP1 foci showed that sOMF caused significant DNA damage in GBM cells and diffuse intrinsic pontine glioma (DIPG) cells but not in normal astroglial SVGp12 cells. Furthermore, sOMF exposure for just 2 h resulted in >40% loss of surviving GBM and DIPG cell colonies detected by clonogenic cell survival assay, similar to that produced by 2 Gy radiation dose. This loss was rescued by the antioxidant Trolox. These results indicate that sOMF stimulation has high anticancer potency comparable to low dose radiation therapy at the cellular level with an underlying mechanism of action that is substantially different from that proposed for Optune® TTF.

Citation Format: Shashank Hambarde, Arvind Pandey, David S. Baskin, Santosh A. Helekar. Spin oscillating magnetic field sOMF of oncomagnetic device selectively kills glioma cancer cells by inducing oxidative stress and DNA damage. [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 4789.
Abstract 4808: Hypoxic HIF-1α stabilization dependent on β3-adrenoceptor localization in Ewing sarcoma
Authors: Pasha Amada , Masi Claudia , Carrozzo Francesco , Rosati Martina , Boaretto Alessia , Pini Alessandro , Favre Claudio , Calvani Maura
Published: April 2023

Hypoxia is a condition of oxygen deficiency which occurs in most growing solid tumors and stimulates a cascade of cell signals through a family of transcription factors named as hypoxia inducible factors (HIFs) that transactivate several regulatory genes involved in tumor proliferation. β3-adrenoceptors (β3-ARs) are involved in several hypoxic scenarios and in pathological conditions where hypoxia leads to important steps for cancer progression. Studies showed that β-AR blockade of mice suppressed hypoxia induction of renal HIF-1α accumulation, erythropoietin production, and erythropoiesis in vivo. In this work we assumed that β3-AR mediates hypoxia sensing and that it was necessary for HIF-1α stabilization thus we investigated a putative correlation between β3-AR/HIF-1α. Data showed a similar outcome of β3-ARs and HIF-1α at short time of hypoxia, and that β3-AR antagonist, SR59230A reduced HIF-1α protein expression under hypoxia. β3-AR silencing under hypoxia revealed an evident reduction of HIF-1α protein expression that confirmed the β3-AR modulation of HIF-1α expression. SR59230A treatment strongly reduced the HIF-1α target genes as GLUT1, HK-2 and HIF-1α expression at long time treatment confirming the β3-AR modulation of HIF-1α transcriptional activity. Conversely, the HIF-1α-transcriptional inhibitor Topotecan did not affect β3-AR expression confirming that is β3-AR that mediates HIF-1α activation. Immunofluorescence analysis showed that β3-AR co-localized with the nucleus under hypoxia more than normoxia. These results were confirmed by nuclear-cytoplasmic fragmentation that showed the presence of both β3-AR and HIF-1α proteins in the nucleus under hypoxia. Co-immunoprecipitation of β3-AR/HIF-1α revealed that under hypoxia, HIF-1α dissociates from β3-AR binding, but this process was abrogated by SR59230A. HIF-1α, released under hypoxia from the link with β3-AR, can play its transcriptional activity in the nucleus, while the binding with β3-AR blocks its transcriptional activity. Moreover, β3-AR regulation on HIF-1α was exerted through PHD2 activity at short time as SR59230A treatment under hypoxia increased the PHD2 protein expression but not at long time treatment, thus showing a different regulation of HIF-1α at different time points. In conclusion, the β3-AR upstream of the signaling β3-AR/HIF-1α axis exerts its role by PHD2 activity.

Citation Format: Amada Pasha, Claudia Masi, Francesco Carrozzo, Martina Rosati, Alessia Boaretto, Alessandro Pini, Claudio Favre, Maura Calvani. Hypoxic HIF-1α stabilization dependent on β3-adrenoceptor localization in Ewing sarcoma. [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 4808.
Abstract 4853: Evaluating the role of melatonin in thyroid cancer cell (MDA-T41) apoptosis and metabolism modulation
Authors: Huang Angela , Sanches Daniel
Published: April 2023

Melatonin, a hormone produced by the pineal gland, is typically known for its modulation of a number of physiological functions, alongside its ability to synchronize the sleep-wake rhythms. In recent years, however, its interactions with cancerous cells and role in prevention and treatment have been increasingly studied. It has been demonstrated that melatonin can improve the efficacy of chemotherapeutic drugs and directly inhibit tumorigenesis in experimental models of neoplasia. In addition, while melatonin typically displays anti-apoptotic effects in normal cells, it holds pro-apoptotic effects in cancer cells. Despite this, the mechanisms by which melatonin affects cell death and metabolism remain unclear due to the seeming difference from cell to cell. We aim to better understand the role of melatonin in regulating thyroid cancer cell apoptosis and metabolism. The MDA-T41 thyroid cancer cell line was cultured with varying quantities of melatonin. Steady volumes of staurosporine (an inductor of apoptotic cell death) were applied to cultures, and cell death levels were subsequently measured using the Realtime-Glo Annexin V Apoptosis and Necrosis assay. Our results showed that melatonin provided anti-apoptotic effects to MDA-T41 cells treated with staurosporine. Both apoptosis and necrosis were reduced by 50% in melatonin-treated cells 24 hours after staurosporine-induced cell death. Our next steps will be to evaluate the mechanisms and further signaling pathways involved in this apoptosis inhibition through Bcl-2 and p53 protein expression.

Citation Format: Angela Huang, Daniel Sanches. Evaluating the role of melatonin in thyroid cancer cell (MDA-T41) apoptosis and metabolism modulation. [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 4853.
Abstract 4911: A novel C-terminal of HSP90 inhibitor NCT-547 eliminates cancer stem-like subpopulation in triple-negative breast cancer
Authors: Jung Eunsun , Jang Seojin , Sung Daeil , Park Soeun , Park Minsu , Ko Dongmi , Kim Seongjae , Seo Juyeon , Nam Kee Dal , Kim Yoon-Jae , Kim Ji Young , Seo Jae Hong , Kang Yong koo , Seock So Ra , Park Jung Min
Published: April 2023

Triple-negative breast cancer (TNBC) harbors a higher cancer stem cell (CSC)-like population and exhibits a more aggressive metastatic phenotype. Due to the limitations of currently available targeted therapies, there remains a significant unmet need for the development of new targeted therapies for TNBC. Molecular chaperone heat shock protein 90 (HSP90) is attracting attention as an ideal therapeutic target due to it regulates essential biological functions such as cell proliferation, angiogenesis and metastasis. Although N-terminal HSP90 inhibitors have had little clinical success to date, C-terminal HSP90 inhibitors have received relatively little attention. We sought to investigate the effects of a new rationally designed NCT-547 on apoptosis, breast cancer stem cell (BCSC)-like properties, migration ability and heat shock response (HSR) in vitro and tumor growth and metastasis in CSC-enriched allograft model in vivo. NCT-547 inhibits TNBC cell proliferation by simultaneously inactivating several survival factors including AKT, MEK, and STAT3. In addition, NCT-547 effectively targets BCSC-like properties with impairment of ALDH1 activity, CD44+/CD24- phenotype, and 3D mammosphere-forming ability. The expression of HSF-1 target genes such as Hsp90, Hsp70, Hsp27, Hsp40 and Hsp65 is highly overexpressed in TNBC patients. The mRNA abundances of target genes were significantly decreased after NCT-547 challenge. NCT-547 effectively targets both the proliferating TNBC tumor cells and CSCs, markedly reducing tumor growth, coinciding with decreased Ki-67 proliferation index and enhanced apoptosis. Anti-tumor effect of NCT-457 was independent of heat shock response as evidenced by significant downregulation of HSF1 phosphorylation and expression of downstream targets HSPs members. It is noteworthy that NCT-547 did not affect markers of hepatic and renal acute toxicity and was not cytotoxic in non-malignant cells. NCT-547 may therefore have potential to address current limitations in the treatment of TNBC.

Citation Format: Eunsun Jung, Seojin Jang, Daeil Sung, Soeun Park, Minsu Park, Dongmi Ko, Seongjae Kim, Juyeon Seo, Kee Dal Nam, Yoon-Jae Kim, Ji Young Kim, Jae Hong Seo, Yong koo Kang, So Ra Seock, Jung Min Park. A novel C-terminal of HSP90 inhibitor NCT-547 eliminates cancer stem-like subpopulation in triple-negative breast 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 4911.
Abstract 4919: Estrogen regulates the immune microenvironment and immunotherapy response of colorectal liver metastases
Authors: Benslimane Yasmine , Lapin Sarah , Chambon Julien , Perrino Stephanie , Brodt Pnina
Published: April 2023

Liver metastases (LM) remain a major cause of cancer-related death and many cancers preferably metastasize to the liver due to its unique anatomical location, rich blood supply and immune-tolerant microenvironment. Up to 50% of colorectal carcinoma patients develop LM during their disease and this is associated with a poor prognosis. Our laboratory previously identified a sexual dimorphism in the regulation of the immune microenvironment (IME) of LM and showed that estrogen could promote the accumulation of myeloid-derived suppressor cells (MDSCs) and Tregs in the liver in response to invading cancer cells. The objective of this study was to elucidate the role of estrogen in the recruitment and polarization of other immune cells and to determine the therapeutic potential of Selective Estrogen Receptor Degrader (SERD) therapy in CRCLM bearing female mice, and evaluate if SERD would improve anti PD-1 immunotherapy efficacy. In estrogen-competent female mice bearing CRCLM, we found increased gene and protein expression of the immunosuppressive cytokine TGF-β, and inversely a decrease in the pro-inflammatory cytokine TNF-alpha. Furthermore, we identified a significant decrease of immunosuppressive CD68+CD163+ M2 macrophages, and an increase of the pro-inflammatory CD68+TNF-alpha+ M1 macrophages in estrogen-depleted (ovariectomized) mice as compared to estrogen-competent mice. This observation was reversed upon estradiol supplementation. Moreover, treatment of female mice with the SERD Fulvestrant, markedly reduced the number of CRCLM compared to vehicle-treated mice, reduced the cell frequencies and counts of M2 macrophages, significantly increased NK cells recruitment in the liver. The addition of SERD to immunotherapy has a better therapeutic effect than both therapies alone, and increased CD8+ T-cells, NK cells, and M1/M2 macrophages ratio. Taken together, our results identify estrogen as a critical regulator of an immunosuppressive TME in the liver and a potential therapeutic target to enhance immunotherapy efficacy.

Citation Format: Yasmine Benslimane, Sarah Lapin, Julien Chambon, Stephanie Perrino, Pnina Brodt. Estrogen regulates the immune microenvironment and immunotherapy response of colorectal liver metastases. [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 4919.
Abstract 4997: PLK1 promotes the metastasis and drug resistance in melanoma
Authors: Mao Fengyi , Kong Yifan , Li Chaohao , Zhang Yanquan , Rao Xiongjian , Zhang Qiongsi , Wang Xinyi , Li Zhiguo , Liu Xiaoqi
Published: April 2023

Melanoma is one of the most frequently diagnosed cancers in the caucasian population of both genders. The survival rate of advanced melanoma patients will be exceedingly poor, as low as 27.3%. BRAF-V600E is the most common mutation found clinically, resulting in the constitutive activation of the MAPK signaling pathway. Vemurafenib, a specific inhibitor of mutant BRAF, has shown an impressive response in phase 3 clinical trials and was approved by FDA in 2011. However, due to the rapid development of resistance, the duration of response under a single treatment is frequently short, highlighting the urgent requirement for novel therapy in melanoma treatment. Polo-like kinase 1 (PLK1), a crucial cell cycle regulator, participates in multiple mitotic processes, including centrosome maturation, mitotic entry, spindle assembly, sister chromatid segregation, mitotic exit, and cytokinesis. Compared to normal tissues, the expression level of PLK1 significantly upregulates in multiple cancers. More importantly, the expression level of PLK1 negatively correlates with the melanoma patients’ survival rate based on the TCGA database. Besides, PLK1 is identified as an oncogene to promote proliferation, motility, and resistance to various drugs. In the present study, we found that overexpression of PLK1 promotes tumor growth, metastasis, metabolic reprogramming, and vemurafenib resistance in the BrafCA/Ptenloxp mouse melanoma model. Through phosphorylation, overexpressed PLK1 stabilizes BACH1, a critical transcriptional factor of multiple metastatic and metabolic genes, and protects it from degradation. Subsequently, a higher level of BACH1 leads to increasing metastasis, upregulation of glycolysis, and suppression of OXPHOs. Besides, we have validated the strong synergy between PLK1 inhibitor volasertib and vemurafenib in treating melanoma carrying BRAF-V600E. Our in vitro and in vivo experiments have shown an improved efficacy of this combined therapy on inhibition of cell proliferation, induction of cell death, and suppression of cell metastasis compared to mono-treatment. In short, our data has indicated PLK1 as a potent and promising target in cancer treatment.

Citation Format: Fengyi Mao, Yifan Kong, Chaohao Li, Yanquan Zhang, Xiongjian Rao, Qiongsi Zhang, Xinyi Wang, Zhiguo Li, Xiaoqi Liu. PLK1 promotes the metastasis and drug resistance in melanoma. [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2023; Part 1 (Regular and Invited Abstracts); 2023 Apr 14-19; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2023;83(7_Suppl):Abstract nr 4997.
Abstract 4999: Preclinical evaluation of MCLA-129, a bispecific antibody targeting EGFR and c-MET on solid tumor cells, in comparison with amivantamab
Authors: de Gorter David J.J. , Deshiere Alexandre , Kramer Arjen , van Rosmalen Martijn , Mortensen Franziska , Geuijen Cecile
Published: April 2023

The HGF/c-MET pathway is frequently upregulated in tumors that are resistant to EGFR tyrosine kinase inhibitors. MCLA-129 is an antibody-dependent cellular cytotoxicity (ADCC) enhanced Biclonics® that targets epidermal growth factor receptor (EGFR) and c-MET in non-small cell lung cancer (NSCLC) and other solid tumors. MCLA-129 is a potent inhibitor of tumor growth applying various mechanisms of action (MOA), including inhibition of c-MET and EGFR signaling, antibody-dependent cellular phagocytosis (ADCP) and ADCC. Here we compared the MOA of MCLA-129 with the MOA of the EGFR-c-MET bispecific antibody amivantamab. The binding of MCLA-129 to tumor cells expressing different levels of EGFR and c-MET was quantified using radiolabeled antibodies. Scatchard plot analysis showed that the affinity of MCLA-129 for both c-MET and EGFR is in the low nanomolar range. In FACS, MCLA-129 and amivantamab show similar binding to tumor cells harboring EGFR or c-MET exon14-skipping mutations. Crossblock experiments with the c-MET binding antibody onartuzumab analog showed that MCLA-129 competes for the same HGF-binding region of c-MET, which is distinct from the binding site of amivantamab according to Neijssen et al, 2021 (PMC8113745). MCLA-129 and amivantamab did not downmodulate EGFR or c-MET. ADCP was also comparable for MCLA-129 and amivantamab, as measured with monocyte-derived macrophages incubated with pHrodo-labeled target cells with readout of tumor cell phagocytosis on the Incucyte® system. In contrast, MCLA-129 displayed ADCC activity similar to or more potently than amivantamab in an ADCC reporter assay with either high affinity FcγRIII 158V-variant or low affinity 158F-variant effector cells in absence or presence of soluble EGFR and soluble c-MET. MCLA-129-induced ADCC appears to be less affected than amivantimab by the presence of soluble EGFR and soluble c-MET. In conclusion, MCLA-129 is a Biclonics® common light chain bispecific antibody with multiple MOA including inhibition of c-MET and EGFR ligand binding, ADCP and ADCC comparable or more potent than amivantamab. A phase 1/2 clinical trial of MCLA-129 in solid tumors is ongoing. These data support the further clinical development of MCLA-129 in patients with NSCLC and other solid tumors.

Citation Format: David J.J. de Gorter, Alexandre Deshiere, Arjen Kramer, Martijn van Rosmalen, Franziska Mortensen, Cecile Geuijen. Preclinical evaluation of MCLA-129, a bispecific antibody targeting EGFR and c-MET on solid tumor cells, in comparison with amivantamab. [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 4999.
Abstract 5014: HPK1 gene knockout reporter cell line: an efficient drug screening system
Authors: Zhai Yunpeng , Li Lidan , Hao Jianyu , Hu Yuqing , Ning Jinying , Hao Feng
Published: April 2023

Immunotherapy is the stimulation of the immune system, improving on the ability of immune system to treat cancer. It is expected to be the most effective therapy for patients with tumor metastasis. Checkpoint inhibitor (CPI) therapy is a form of cancer immunotherapy which can block inhibitory checkpoints, restoring immune system function. Tumor cells could recruit immunomodulatory cells and induce inhibitory signals, resulting in losing immunomodulatory. Therefore, there is an urgent need to develop new therapeutic agents that work in concert with CPI. HPK1, also known as MAP4K1, can act as a negative regulator of activation signals generated by T cell receptor (TCR). The development and screening of HPK1 inhibitors have great potential in the cancer immunotherapy. Herein, we constructed a HPK1 knockout reporter cell line Jurkat-NFAT-Luc2-HPK1-Knockout-Cell-Line by CRISPR/Cas9 which can be used in high-throughput HPK1 inhibitors screening. Wild type cells were incubated with HPK1 inhibitors and luciferase signals were compared in HPK1-knockout cells. Our HPK1 knockout reporter cell line has high sensitivity and specificity. The reporter system can achieve high-throughput compound library screening. After selected the compound with selective inhibitory activity, further validation and optimization would be performed. Moreover, we constructed a Jurkat-HPK1-HiBiT reporter cell line by CRISPR/Cas9. It can be used in the screening of HPK1 degradants. We believe that HPK1 gene editing cell lines play an important role in the high-throughput screening of drugs. It could facilitate the development of drugs in tumor immunotherapy.

Citation Format: Yunpeng Zhai, Lidan Li, Jianyu Hao, Yuqing Hu, Jinying Ning, Feng Hao. HPK1 gene knockout reporter cell line: an efficient drug screening system. [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 5014.
Abstract 5052: IB-DNQ enhances radiation induced ferroptosis in NQO1-positive cancer cells
Authors: Huang Xiumei , Su Xiaolin , Boudreau Matthew W. , Hergenrother Paul J.
Published: April 2023

Cancer is the second leading cause of death worldwide. Current treatments for solid cancers are associated with inherent resistance mechanisms and are ineffective against non-cycling cancer cells. Thus, there is a critical need to develop novel tumor-selective combination therapies to overcome resistance while optimally spare normal tissues. In this study, we investigate the mechanistic basis for tumor-selective potentiation of ionizing radiation (IR) in combination with IB-DNQ, a novel NQO1-bioactivatable agent. We found that combination of a low dose of ionizing radiation (IR, 2 Gy) with a sublethal dose of IB-DNQ dramatically increased Fe2+ leakage and induced elevated lipid peroxidation and ferroptosis in NQO1-positive cancer cells via upregulating the expression of ACSL4, a lipid metabolism enzyme required for ferroptosis. Knockdown of endogenous ACSL4 significantly inhibited lipid peroxidation via downregulating the expression of PTGS2 and dramatically inhibited the cell death that induced by IR or IB-DNQ alone or combination treatment. We further found that this combination treatment significantly decreased IR or IB-DNQ alone mediated upregulation of the expression of GPX4 that led to the accumulation of lipid peroxides, which suggests that IR or IB-DNQ-induced expression of GPX4 may function as a negative feedback loop to restore cell survival upon a sublethal dose of IR or IB-DNQ treatment. Finally, we demonstrated that this combination therapy synergistically suppressed tumor growth and dramatically extended mouse lifespan in orthotopic pancreatic and non-small-cell lung cancer xenograft models. Together, our study provides novel preclinical evidence for new combination therapy in NQO1-positive solid tumors that may overcome IR-induced resistance.

Citation Format: Xiumei Huang, Xiaolin Su, Matthew W. Boudreau, Paul J. Hergenrother. IB-DNQ enhances radiation induced ferroptosis in NQO1-positive 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 5052.
Abstract 5183: Development of an aerosol-based immunotherapy for lung cancer
Authors: Li Yongren , Zhao Zhen , Fetse John , Hasan Reaid , Mamani Umar-Farouk , Guo Yuhan , Cheng Kun
Published: April 2023

Lung cancer is one of the leading causes of cancer death and the second most cancer in the world. Compared to systematic administration, local delivery of therapeutic agents to the lung increases their accumulation in lung cancer cells and reduces the toxicity in other organs. We previously discovered an anti-PD-L1 peptide that can be potentially uses for lung cancer therapy. This project aims to develop an aerosolized formulation of the anti-PD-L1 peptide for the treatment of lung cancer. We hypothesize that the aerosolized peptide has higher accumulation in the lung compared to systematic formulations, leading to high therapeutic index with less side effects. We developed a spray freeze-drying procedure and optimized the formulation to prepare the peptide dry powders. The median mass aerodynamic diameter (MMAD) and geometric standard deviation (GSD) of the aerosols were determined by an 8-stage Andersen Cascade Impactor. The morphology of the aerosols was studied using a scanning electron microscope (SEM). The stability of the aerosols in lung fluid was evaluated. Blocking assay suggested that the aerosolized formulation maintains the blocking efficiency of the peptide. Intratracheal administration of the peptide dry powder shows a high accumulation of the peptide in the lung.

Citation Format: Yongren Li, Zhen Zhao, John Fetse, Reaid Hasan, Umar-Farouk Mamani, Yuhan Guo, Kun Cheng. Development of an aerosol-based immunotherapy for 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 5183.
Abstract 5196: Novel humanized CD36 mouse model for therapeutic studies
Authors: Wang Linlin , Liu Chang , Soto Rebecca , Shang Chengzhang , Lin Qingcong
Published: April 2023

Activation of oncogenic signaling pathways can be mediated by various transporters on tumor cells that facilitate trafficking of lipids. CD36 has been identified as a fatty acid transporter and participates in tumor growth and drug resistance. To explore potential therapeutic strategies targeting CD36, we generated a human CD36 knockout/knock-in in situ (B-hCD36) mouse model to evaluate the efficacy of anti-human CD36 antibodies. We first confirmed human CD36 gene and protein expression in B-hCD36 mice by RT-PCR and flow cytometry, respectively. We next observed that the overall development, differentiation, and distribution of splenic, blood and lymph node immune cells were similar between wild-type C57BL/6 and B-hCD36 mice. Lastly, in vivo efficacy studies using anti-human CD36 antibodies alone or in combination with anti-mouse PD-1 antibodies, were effective in controlling MC38 tumor growth in B-hCD36 mice. Our data demonstrates that B-hCD36 mice are a powerful preclinical model for in vivo efficacy evaluation of anti-human CD36 antibodies.

Citation Format: Linlin Wang, Chang Liu, Rebecca Soto, Chengzhang Shang, Qingcong Lin. Novel humanized CD36 mouse model for therapeutic studies. [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 5196.
Abstract 5319: EU-OPENSCREEN - An open-access initiative in chemical biology to support anti-cancer drug discovery
Authors: Stechmann Bahne , Wit Charlotte
Published: April 2023

Over the past two decades, an increasing number of small molecule drugs that target unique molecular changes underlying a specific cancer have been approved, and small molecule drugs represent a major therapeutic class for cancer treatment. Chemical ‘probes’ are also indispensable tools to advance our understanding of pathologies at the molecular and cellular level and to validate novel drug targets.

A major bottleneck in identifying and developing these chemical compounds in academia is often access to robust disease-relevant assays, high-throughput screening platforms, comprehensive screening libraries, medicinal expertise for the hit-to-lead optimization. In order to democratize access to these resources for a broader scientific community, the publicly funded EU-OPENSCREEN (www.eu-openscreen.eu) initiative has been established with the aim to accelerate drug discovery efforts in an open-access setting in collaboration with researchers from academia and industry.

EU-OPENSCREEN’s 30 expert centers offer access to their cancer target-based and phenotypic drug screening expertise, capabilities in developing disease-relevant assays and hit-to-lead optimization capabilities. The identified bioactive ‘hit’ compounds will be progressed into high-quality chemical starting points to be further developed into drug leads or chemical probes for the elucidation of mechanisms of cancer onset, progression, resistance and effects of drug therapy. All generated tool compounds and associated bioactivity data are made available to the global scientific community.

Here we will present an overview of anti-cancer drug discovery projects of EU-OPENSCREEN and explain the opportunities and benefits for other international scientists to collaboratively develop novel anti-cancer drug leads and chemical probes.

EU-OPENSCREEN receives funding from the European Union's Horizon 2020 and Horizon Europe research and innovation programs under grant agreements No 654248 and 101058620.

Citation Format: Bahne Stechmann, Charlotte Wit. EU-OPENSCREEN - An open-access initiative in chemical biology to support anti-cancer drug discovery. [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 5319.
Abstract 5333: Combining chemoproteomics with machine learning identifies functionally active covalent fragments for hard-to-drug cancer drivers
Authors: Hermann Johannes C. , Everley Robert , Marholz Laura , Berberich Matthew , Yang Tzu-Yi , Chao Yu-Hsin , Awol Abduselam K. , Shaghafi Michael , Yoon Han , Varma Rohan , Stein Reed , Krug Karsten , Lachtara Emily , Erlanson Daniel , Varma Chris , Webster Kevin R.
Published: April 2023

a) Many cancer drivers are considered “undruggable” and without targeted treatments because they lack binding sites for conventional small molecules. Here, we introduce The FRONTIER™ Platform applying machine learning (ML), chemoproteomics and covalent chemistry to identify binding sites and cell-active covalent fragments across the human proteome, including against most cancer drivers and previously “undruggable” targets. Molecules tested in functional assays are active and can serve as starting points for new drug discovery initiatives.

b) We are using mass spectrometry and data analysis workflows to perform high-throughput chemoproteomic profiling experiments. These experiments identify hits across the proteome using different cancer-relevant cell backgrounds and characterize binding sites for drug discovery. Customized ML algorithms using chemoproteomic, genomic and structural data to characterize and prioritize identified binding sites for covalent drug discovery. The performance of the platform allows the profiling of thousands of compounds from a custom-built library that has been optimized by ML for covalent fragment-based drug discovery. The nature of the fragment hits and the ability to map them to and focus on preferred binding sites for covalent drugs enables accelerated lead generation.

c) We show details of the platform highlighting library design concepts and the hotspot map with binding site prioritization algorithms. We show coverage and applicability across important cancer target classes and signaling pathways. Covalent fragment hits have been identified for multiple difficult cancer targets, including KRAS, p53, STAT3, KEAP1, PTPN11 and others. The platform also identifies ligands for novel allosteric binding sites in established oncology targets such as CDK4, PI3KCA and BTK. We will highlight how discovered ligands show functional activity in orthogonal assays, demonstrating their fitness for drug discovery campaigns and target validation experiments.

d) Undruggable targets across a variety of cancer target classes have become druggable.

Citation Format: Johannes C. Hermann, Robert Everley, Laura Marholz, Matthew Berberich, Tzu-Yi Yang, Yu-Hsin Chao, Abduselam K. Awol, Michael Shaghafi, Han Yoon, Rohan Varma, Reed Stein, Karsten Krug, Emily Lachtara, Daniel Erlanson, Chris Varma, Kevin R. Webster. Combining chemoproteomics with machine learning identifies functionally active covalent fragments for hard-to-drug cancer drivers. [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 5333.
Abstract 5343: Evaluating the screened polyphenolics molecules as potential chemosensitizers against the oncogenic signalling proteins in lung cancer
Authors: Arora Swati , Sheoran Sumit , Basu Tanmayee , Subbarao Naidu , Upadhyay Atul Kumar , Vuree Sugunakar
Published: April 2023

Background: Lung cancer is the most commonly diagnosed cancer globally. There are numerous agonists and antagonistic treatment interventions available for therapy. However, all of them possess different detrimental consequences in addition to resistance and remission in sufferers. Medicinal plants have traditionally been employed to treat a wide range of ailments and problems.

Methodology: A total of 550 flavonoids and small molecules retrieved from literature and databases (Chembl, and PubChem) were analysed for molecular docking, biological activities, ADME, and drug-like properties through SwissADME and PreADMET, Molecular Simulation, and cytotoxicity studies. Simultaneously, review literature is employed to identify the novel protein targets viz. ROS1 (3ZBF) & EGFR (6DUK). Additionally, the crystal structures of the proteins were obtained from PDB and their domains and active sites were predicted using ScanProsite, MOTIF Search, metaPOCKET2, and PrankWeb tools. Finally, the proteins and ligands were energy-minimized using Chimera and open babel tool and subjected to virtual screening and molecular docking using GOLD. Furthermore, the molecular simulation was performed at 100ns by utilizing GROMACS, and cytotoxicity studies were performed by employing the A549 Lung cancer cell line.

Results: Diosmin (64.57) and Hesperidin (60.98) had the greatest binding affinity with the identified targets, i.e., ROS1 (3ZBF) and EGFR(6DUK) respectively against Lung Cancer, and remained stable during 100ns of molecular simulation. Furthermore, we have performed cytotoxicity studies on A549 cell lines that show both the compounds have significant anticancer activity and do not affect healthy cells in the range of, i.e., Diosmin (500 µg/ml to 0.024 µg/ml.) and Hesperidin (500 µg/ml to 125 µg/ml).

Conclusion: After the analyses, we can conclude that it has the potential to be exploited as a novel therapeutic agent in the treatment of lung cancer.Keywords: Diosmin, Hesperidin, In-silico, A549, Lung Cancer, Simulation

Citation Format: Swati Arora, Sumit Sheoran, Tanmayee Basu, Naidu Subbarao, Atul Kumar Upadhyay, Sugunakar Vuree. Evaluating the screened polyphenolics molecules as potential chemosensitizers against the oncogenic signalling proteins in 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 5343.
Abstract 5432: Validation of spatial gene expression patterns predicted by deep convolutional neural networks from breast cancer histopathology images
Authors: Ton Kathy , Wang Yinxi , Pan Liuliu , Kartasalo Kimmo , Acs Balazs , Weitz Philippe , Zhang Liang , Liang Yan , Hartman Johan , Volkonen Masi , Larsson Christer , Ruusuvuori Pekka , Beechem Joseph , Rantalainen Mattias
Published: April 2023

Molecular profiling is central in cancer precision medicine but remains costly and is only based on tumor average profiles. Morphologic patterns observable in histopathology sections from tumors are determined by the underlying molecular phenotype and therefore have the potential to be exploited for the prediction of molecular phenotypes. Transcriptome-wide expression morphology (EMO) analysis with deep convolutional neural networks (CNN) enables the prediction of mRNA expression and proliferation markers from routine histopathology whole slide images in breast cancer. The NanoString GeoMx® Digital Spatial Profiler (DSP) platform has capabilities of associating crucial spatial information with intratumor variabilities of gene expression, serving as an ideal tool to characterize and validate the prediction of intratumor heterogeneity. The GeoMx platform also provides functionalities of overlaying and aligning hematoxylin and eosin (H&E) whole slide scans on serial tissue sections with morphology marker staining to facilitate region of interest (ROI) selection to match relevant image tiles from the model input, which is a key step to ensure accuracy for the evaluation of the model predictions. The GeoMx RNA Immune Pathways Panel contains 84 gene targets including key genes involved in immune pathways and tumorigenesis and was used to validate the prediction of mRNA expression output from deep CNN models.

Citation Format: Kathy Ton, Yinxi Wang, Liuliu Pan, Kimmo Kartasalo, Balazs Acs, Philippe Weitz, Liang Zhang, Yan Liang, Johan Hartman, Masi Volkonen, Christer Larsson, Pekka Ruusuvuori, Joseph Beechem, Mattias Rantalainen. Validation of spatial gene expression patterns predicted by deep convolutional neural networks from breast cancer histopathology images. [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 5432.
Abstract 5494: Preclinical combination of ONC201 with radiotherapy and Temozolomide in a GBM mouse orthotopic model results in reduced tumor burden and prolonged survival
Authors: Zhou Lanlan , Wu Laura Jinxuan , Zhang Jun , George Andrew , Hahn Marina , Zhang Leiqing , Seyhan Attila A. , El-Deiry Wafik S.
Published: April 2023

Glioblastoma (GBM) is the most common and lethal primary malignancy of the central nervous system. It is estimated that more than 13,000 new cases of GBM will be diagnosed this year in the United States. Despite multidisciplinary treatments such as surgery, chemotherapy, and radiotherapy, the five-year survival rate for GBM patients is only 6.8 percent and has shown no notable improvement in the last three decades. There have only been five drugs and one device ever approved by the FDA for the treatment of GBM since it was first identified in the scientific literature in the 1920’s. Our in vitro studies suggested that first-in-class small-molecule imipridone TIC10/ONC201 can inactivate ERK/AKT, induce the integrated stress response (ISR), upregulate pro-apoptotic TRAIL receptor DR5, deplete cancer stem cells, and induce mitochondrial dysfunction, growth arrest or cell death in GBM cells. Knockdown of ClpP protects GBM cells from ONC201 but not TMZ. ONC201 crosses the blood-brain barrier and has induced durable tumor regressions in adult and pediatric H3K27M-mutant diffuse midline glioma patients. We hypothesized that ONC201 may synergize with radiotherapy and temozolomide in GBM. GBM mouse orthotopic models were established through intracranial injection of luciferase expressing U251 GBM cells with a KOPF model 940 small animal stereotaxic frame and a Stoelting Quintessential Stereotaxic Injector. Tumor formation and growth was confirmed with bioluminescence imaging. Randomized treatment group mice received weekly treatment of ONC201 (100 mg/kg p.o.) and/or radiotherapy (2 Gy local irradiation) and/or TMZ (20 mg/kg i.p.) for four weeks for long term survival and tumor monitoring or one week for short term biomarker studies. We observed that the triple combination of ONC201, radiotherapy and TMZ significantly prolongs survival and reduces tumor burden compared to single treatment and dual combinations. Short term biomarker studies demonstrated that triple combination treatment decreases tumor cell proliferation, induces more apoptosis and inhibits ClpX to unleash mitochondrial ClpP. Our data support further development of the triple combination regimen of ONC201, TMZ and radiation therapy for GBM first-line treatment.

Citation Format: Lanlan Zhou, Laura Jinxuan Wu, Jun Zhang, Andrew George, Marina Hahn, Leiqing Zhang, Attila A. Seyhan, Wafik S. El-Deiry. Preclinical combination of ONC201 with radiotherapy and Temozolomide in a GBM mouse orthotopic model results in reduced tumor burden and prolonged survival. [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 5494.
Abstract 5630: Robust spatial biomarker discovery through multi-platform multiplex image analysis of breast cancer clinical cohorts
Authors: Eng Jennifer R. , Bucher Elmar , Hu Zhi , Sanders Melinda , Chakravarthy Bapsi , Pietenpol Jennifer , Sears Rosalie C. , Gibbs Summer , Gray Joe W. , Chin Koei
Published: April 2023

Spatial profiling of tissues promises to elucidate tumor-microenvironment interactions and enable development of spatial biomarkers to predict response to immunotherapy and other therapeutics. However, spatial biomarker discovery is often limited to small patient cohorts and single technologies, limiting statistical power and increasing the likelihood of technical artifacts. We utilized cyclic immunofluorescence, (CyCIF), to profile patient tissues from 102 breast cancer patients, 63 with clinical follow up. We then developed methods for comparative analysis of data from three disparate imaging technologies including our CyCIF images, as well as publicly available imaging mass cytometry and multiplex ion-beam imaging breast cancer data sets. We demonstrate similar single-cell phenotyping results across breast cancer patient cohorts imaged with the three methods, and furthermore, show agreement in the prognostic value of cellular and spatial biomarkers across platforms. We identified T cell infiltration as independently associated with longer survival in high-proliferation breast cancer, which was enriched for activated and spatially clustered T cells. A comparison of six spatial analysis methods revealed robust spatial biomarkers, including tumor-macrophage and tumor-fibroblast proximity associated with poor prognosis in estrogen receptor positive and triple negative tumors, respectively. Our methods enable assembly of larger clinical cohorts from diverse platforms to aid in predictive and prognostic spatial biomarker identification and validation.

Citation Format: Jennifer R. Eng, Elmar Bucher, Zhi Hu, Melinda Sanders, Bapsi Chakravarthy, Jennifer Pietenpol, Rosalie C. Sears, Summer Gibbs, Joe W. Gray, Koei Chin. Robust spatial biomarker discovery through multi-platform multiplex image analysis of breast cancer clinical cohorts. [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 5630.
Abstract 5834: The pro-tumoral function of cancer-associated mesothelial cells
Authors: Chauvin Maeva
Published: April 2023

Cancer-associated mesothelial cells have emerged as significant contributors to the tumor microenvironment, influencing tumor growth and immune evasion. Mesothelial cells are the first point of contact with ovarian cancer cells during peritoneal spread. Peritoneal mesothelial cells also play an essential role in immune surveillance. However, the mechanism by which cancer and mesothelial cells within the tumor microenvironment coordinate growth and immune evasion remains poorly understood. To study the role of mesothelial cells in the tumor microenvironment, we have recently generated novel syngeneic mouse models of high-grade serous ovarian cancer, which were derived from Fallopian tube epithelium, and bear human-relevant mutational genotypes. Analysis of one such model (CCNE1 amplified) using single-cell transcriptomics in omental metastases revealed specific expression of the anti-Mullerian hormone type II receptor (Amhr2) in cancer-associated mesothelial cells, whereas normal omental mesothelium does not express Amhr2. Co-culture of cancer cells with normal omental mesothelial cells induces AMHR2 expression along with a pro-tumorigenic cancer-associated mesothelial cell phenotype. Overexpression of AMHR2 in a human immortalized mesothelial cell line significantly modulated the expression of genes involved in immune response pathways, remodeling of extra-cellular matrix, and secretion of growth factors by RNAseq and qPCR. Furthermore, we showed that anti-Mullerian hormone (AMH) (the ligand of AMHR2), is secreted by ovarian cancer cells and further downregulates the chemoattractant cytokines secreted by CAMCs, thus promoting immunosuppression within the tumor. Finally, implantation of ovarian cancer cells in syngeneic Amhr2-/- hosts results in tumors that grow significantly slower than those implanted in wild-type hosts. Together these data suggest AMHR2 is a specific marker of CAMCs, whose expression is necessary for the reprogramming of mesothelial cells into CAMCs. The AMH-AMHR2 axis mediates paracrine signaling between cancer cells and CAMCs within the tumor microenvironment to promote tumor growth and immune evasion. These results suggest that AMHR2 could be a highly specific target to enhance immunotherapies in ovarian cancer.

Citation Format: Maeva Chauvin. The pro-tumoral function of cancer-associated mesothelial 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 5834.
Abstract 5843: miR-224 activates cancer-associated fibroblasts to promote cancer cell invasion in lung tumor microenvironment
Authors: Oh Seonyeong , Cheon Inyoung , Ahn Young-Ho
Published: April 2023

Cancer-associated fibroblasts (CAFs) promote EMT (epithelial-mesenchymal transition), invasion, and metastasis of cancer cells in the tumor microenvironment. To identify microRNAs (miRNAs) associated with the reprogramming of normal lung fibroblasts (LFs) into CAFs, global miRNA expression was profiled in LFs and CAFs through Nanostring nCounter. miR-224 was one of the increased miRNAs in CAFs, and its expression was higher in tumors than in normal tissues in dbDEMC and TCGA_lung adenocarcinoma data. miR-224 levels were then checked by qRT-PCR in paraffine-embedded tumor samples from patients with lung adenocarcinoma. Kaplan-Meier survival analysis showed that high miR-224 expression was related to poor survival. These data show that lung cancer progression is promoted by miR-224. To discover the roles of increased miR-224 in CAFs, miR-224 was overexpressed by lentiviral infection in LFs. LF_miR-224 overexpression enhanced the expression of CAF activation markers including Fap, S100a4, and Vim. In spheroid invasion assay using co-culture of LFs and cancer cells, as an experimental model of the tumor microenvironment, LF_miR-224 promoted invasion of lung cancer cells compared with control LFs. In transwell migration assay, LF_miR-224 stimulated migration of lung cancer cells more than control LFs. Collectively, these results indicate that CAFs reprogrammed by miR-224 promote the migration and invasion of lung cancer cells.

Citation Format: Seonyeong Oh, Inyoung Cheon, Young-Ho Ahn. miR-224 activates cancer-associated fibroblasts to promote cancer cell invasion in lung tumor microenvironment. [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2023; Part 1 (Regular and Invited Abstracts); 2023 Apr 14-19; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2023;83(7_Suppl):Abstract nr 5843.