bluuebunny/crossref_metadata_2025_split huggingface dataset:
Abstract 3970: Tumor-associated macrophage, angiogenesis and lymphangiogenesis predicts prognosis in patients with non-small cell lung cancer
Authors: Hwang Ilseon , Park Keun Uk , Jung Eun Hee , Ryoo Hun-Mo , Ylaya Kris , Chung Joon-Yong , Hewitt Stephen M. 
Published: September 2020

Abstract
Background: The tumor microenvironment (TME) is a critical player in tumor progression, metastasis and therapy outcome. Tumor-associated macrophages (TAMs) are a well-recognized core element of the TME, and are generally characterized as M2-like macrophages. TAMs are believed to contribute to tumor progression, but the mechanism behind this remains unclear. We aimed to investigate the clinical value, angiogenic and lymphangiogenic significance of TAMs in non-small cell lung cancer (NSCLC).
Methods: We assessed CD68, CD163, VEGF-A, and VEGF-C expression in 349 patients with NSCLC by combined immunohistochemistry and digital image analysis. Subsequently, the potential association between M2 TAMs and angiogenetic VEGF-A and/or lymphangiogenetic VEGF-C was evaluated for its prognostic value. Furthermore, the effects of M2 TAMs on angiogenesis and lymphangiogenesis were explored in vitro co-culture system.
Results: Elevated M2 (CD163+/CD68+) ratio was significantly associated with poor overall survival (p=0.023). Dual high expression of M2 ratio and VEGF-C was correlated with worse overall survival (p=0.033). Furthermore, CD68 and CD163 expression are positively associated with VEGF-A and/or VEGF-C expression. Multivariate analysis revealed that M2 ratio was an independent predictor of poor survival [HR (95% CI) = 1.53 (1.01-2.33), p=0.046]. Combined M2+/VEGF-C+ expression was also an independent predictor of poor survival [HR (95% CI) = 2.01 (1.28-3.16), p=0.003]. Notably, we confirmed that M2 macrophages enhanced the protein and mRNA expression of both VEGF-A and VEGF-C significantly, while M1 macrophages induced only mRNA expression of VEGF-A significantly in A549 cells.
Conclusions: This study suggests that TAMs are significantly associated with angiogenesis or lymphangiogenesis in the progression of NSCLC. Furthermore, elevated M2 ratio and combined M2 and VEGF-C high expression are strong indicators of poor prognosis in patients with NSCLC.
Citation Format: Ilseon Hwang, Keun Uk Park, Eun Hee Jung, Hun-Mo Ryoo, Kris Ylaya, Joon-Yong Chung, Stephen M. Hewitt. Tumor-associated macrophage, angiogenesis and lymphangiogenesis predicts prognosis in patients with non-small cell lung cancer [abstract]. In: Proceedings of the Annual Meeting of the American Association for Cancer Research 2020; 2020 Apr 27-28 and Jun 22-24. Philadelphia (PA): AACR; Cancer Res 2020;80(16 Suppl):Abstract nr 3970.
Abstract 4018: Interaction site elucidation of the Rac1/Cdc42 inhibitor, MBQ-167
Authors: Medina Julia I. , Bayro Marvin , Hernandez Eliud , Vlaar Cornelis , Gonzalez Ricardo , Dharmawardhane Suranganie 
Published: September 2020

Abstract
Metastasis continues to be the primary cause of cancer-related death. Therefore, we focus on designing small molecules inhibitors to block metastasis. The Rho GTPase family members Rac1, and Cdc42 are critical regulators of cancer cell migration and invasion, and thus, metastasis. Rac1 is activated by Guanine Nucleotide Exchange Factors (GEFs), which promote the exchange of GDP to GTP. In most cancers, the homologous Rac1 and Cdc42 are hyperactivated due to the overexpression of GEFs. Therefore, we designed inhibitors to block the activation and interaction of Rac1 and Cdc42 with their GEFs. Our lead compound MBQ-167, is a dual inhibitor of Rac1 and Cdc42 with IC50s of 103 nM and 78 nM, respectively. MBQ-167 inhibited the viability and migration of metastatic cancer cells without affecting non-metastatic or non-cancer cells. Moreover, MBQ-167 reduced mammary tumor growth and metastasis of HER2 positive and triple-negative breast cancer cells in immunocompromised mice by ~ 90%. To further elucidate the mechanism of action of MBQ-167, we tested the hypothesis that MBQ-167 blocks the interaction of specific Rac1/Cdc42 GEFs by interaction with particular amino acid residues in the switch I and II homologous regions of Rac1 and Cdc42 proteins. Therefore, to achieve the objective of elucidating the interaction site of MBQ-167 on Rac1 and Cdc42, we used in silico analysis and Nuclear Magnetic Resonance (NMR) of MBQ-167 with Rac1 in a GDP bound state. These results were biochemically validated by pulldown assays using a G15A nucleotide-free mutant predicted to interact tightly with Rac1/Cdc42 GEFs. In silico, using Autodock Tools, MBQ-167 was docked to the GEF binding domain of Rac1 and Cdc42 with low binding energy, indicating a tight interaction. For the NMR structural analyses, Rac1 was isotopically labeled with 15N and titrated with several concentrations of MBQ-167, at ratios of 1:1, 1:2, 1:3, and 1:4. We were able to detect chemical shift perturbations in key residues in the switch I and II regions that interact with Rac1/Cdc42 GEFs and regulate the activity. Moreover, these perturbations were more intense with increasing amounts of MBQ-167, confirming the specificity of the interaction. Therefore, our data signify the direct interaction of MBQ-167 with Rac1, thus validating the role of MBQ-167 as a specific Rac1/Cdc42 inhibitor.
Citation Format: Julia I. Medina, Marvin Bayro, Eliud Hernandez, Cornelis Vlaar, Ricardo Gonzalez, Suranganie Dharmawardhane. Interaction site elucidation of the Rac1/Cdc42 inhibitor, MBQ-167 [abstract]. In: Proceedings of the Annual Meeting of the American Association for Cancer Research 2020; 2020 Apr 27-28 and Jun 22-24. Philadelphia (PA): AACR; Cancer Res 2020;80(16 Suppl):Abstract nr 4018.
Abstract 4027: Metformin treatment inhibits uterine leiomyoma and leiomyosarcoma cells growth and migration in vitro
Authors: Ricci Anamaria R. , Maffazioli Giovana D. , Baracat Edmund C. , Carvalho Kátia C. 
Published: September 2020

Abstract
Leiomyoma (LM) and leiomyosarcoma (LMS) are uterine mesenchymal tumors that present variable clinical and biological behavior. They are associated with infertility, pelvic pain and abnormal uterine bleeding.. LM is a benign tumor commonly affecting women at reproductive age whereas LMS represents 40% of all uterine sarcomas. These tumors do not have specific therapy and their origin is still uncertain. Some researchers believe that degenerated LM may to turn to LMS. FOXO3a, a transcriptional factor, has been associated to regulation of several cellular functions as well as tumors development. Previously, we detected that FOXO3a gene and protein expression present different expression profiles in LM and LMS. In this current study, our aim was to investigate the in vitro effects of metformin and genistein, both inhibitors of AKT-mediated FOXO3a phosphorylation, treatment in LM and LMS cells behavior. LM (THESCs CRL-4003) and LMS (SK-UT-1) cell lines were grown in specific mediums and conditions (ATCC recommendations), and treated with different drugs concentrations (0.1 - 200 uM and 0.1 - 200 nM, for metformin and genistein, respectively) at several time points (0, 24, 48, 72, 96 and 120 h). After treatment, cells were evaluated for gene and protein expression, cell proliferation, migration and apoptosis. Gene expression was assessed by quantitative Real Time PCR and proteins were evaluated by immunocitochemical analysis. Genistein depicted high toxicity for the cells, due to its vehicle (DMSO), in all tested dose and times. Metformin presented higher efficiency in cell proliferation and migration inhibition for both cell lines, as well as in the apoptosis induction. Prominent effects were observed in LMS cells after treatment with 25 uM for 72h. Gene expression analysis showed a slight decrease in FOXO3a expression after treatment with these drugs. Our results suggest that treatment with metformin induces the FOXO3a function restoration in the cells, by inhibition of the AKT/PI3K pathway, and can represent a future alternative target therapy for these neoplasias.
Citation Format: Anamaria R. Ricci, Giovana D. Maffazioli, Edmund C. Baracat, Kátia C. Carvalho. Metformin treatment inhibits uterine leiomyoma and leiomyosarcoma cells growth and migration in vitro [abstract]. In: Proceedings of the Annual Meeting of the American Association for Cancer Research 2020; 2020 Apr 27-28 and Jun 22-24. Philadelphia (PA): AACR; Cancer Res 2020;80(16 Suppl):Abstract nr 4027.
Abstract 4029: BET inhibitor suppresses neutrophil-dependent lung metastasis of inflammatory renal cancer cells
Authors: Ehata Shogo , Nishida Jun , Miyakuni Kosuke , Koinuma Daizo , Miyazono Kohei 
Published: September 2020

Abstract
Recent studies have shown that cancer cells alter the immune landscape by secreting inflammation-related cytokines or chemokines through a process known as cancer cell-intrinsic inflammation. Cancer cell-intrinsic inflammation facilitates both the initial progression of cancers and cancer metastasis via tumor microenvironment reconstitution, immune surveillance, and premetastatic niche formation. However, in the case of clear cell renal carcinoma (ccRCC), the role of cancer cell-intrinsic inflammation is not fully uncovered. In the present study, we explored the function and regulatory mechanisms of cancer cell-intrinsic inflammation in ccRCC.
First, highly malignant derivatives were obtained through the repetitive exposure of human ccRCC cells into mouse renal microenvironment. Of note, these derivatives elicited cancer cell-intrinsic inflammation in the host mice, which facilitated lung metastasis in neutrophil-dependent manner. Thus, we defined these derivatives as inflammatory ccRCC cells and characterized in the following study.
Next, the regulatory mechanisms for the transcription of genes related to neutrophil dynamics were explored. RNA-sequencing analysis revealed that mRNA-expression levels of four CXC chemokines were up-regulated in all inflammatory ccRCC derivatives. ChIP-sequencing analysis of H3K27ac or bromodomain-containing 4 (BRD4) indicated that super enhancers were recruited at each gene in inflammatory ccRCC derivatives.
Finally, we evaluated the effect of the bromodomain and extra-terminal motif (BET) inhibitor on neutrophil-dependent lung metastasis of inflammatory ccRCC cells. Although the sizes of primary tumors in inflammatory ccRCC derivative-bearing mice were not altered by the treatment of BET inhibitor, those of metastatic lung tumors were significantly decreased.
Overall, our findings revealed a functional role of cancer cell-intrinsic inflammation, which primes neutrophils and drives metastasis of ccRCC cells. The primary tumor microenvironment enriched inflammatory ccRCC cells via epigenetic remodeling. We also showed the utility of inflammation management in suppressing metastasis by utilizing a BET inhibitor.
Citation Format: Shogo Ehata, Jun Nishida, Kosuke Miyakuni, Daizo Koinuma, Kohei Miyazono. BET inhibitor suppresses neutrophil-dependent lung metastasis of inflammatory renal cancer cells [abstract]. In: Proceedings of the Annual Meeting of the American Association for Cancer Research 2020; 2020 Apr 27-28 and Jun 22-24. Philadelphia (PA): AACR; Cancer Res 2020;80(16 Suppl):Abstract nr 4029.
Abstract 4044: Pharmacological targeting of the circadian clock, a new frontier for cancer therapy
Authors: Sulli Gabriele , Rich Jeremy 
Published: September 2020

Abstract
The circadian clock imposes daily rhythms in cell proliferation, metabolism, inflammation and DNA damage response. Perturbations of these processes are hallmarks of cancer. This raises the hypothesis that pharmacological modulation of the circadian machinery may be an effective therapeutic strategy for combating cancer. Indeed, targeting a circadian regulator could affect multiple cancer vulnerabilities, thus mimicking combination therapy. Here we show that pharmacological modulation of clock genes is specifically lethal to cancer cells and oncogene-induced senescent cells and have no effect on the viability of normal cells or tissues. This novel strategy can target multiple oncogenic drivers (such as HRAS, BRAF, PIK3CA, lack of p53 and others) and is effective under hypoxic conditions. Autophagy and de novo lipogenesis two crucial metabolic nodes for cancer survival are simultaneously blocked by chemical clock modulators. Notably, this approach impairs glioblastoma growth in vivo and improves survival without causing overt toxicity in mice. These results indicate that pharmacological modulation of circadian regulators is a novel antitumor strategy, identifying a class of anticancer agents with a wide therapeutic window. This new therapeutic paradigm could offer promising tools for the treatment of highly heterogeneous tumors that are unaffected by targeted therapy. The integration of circadian biology into cancer research therefore offers new options for making cancer treatment more effective and selective.
Citation Format: Gabriele Sulli, Jeremy Rich. Pharmacological targeting of the circadian clock, a new frontier for cancer therapy [abstract]. In: Proceedings of the Annual Meeting of the American Association for Cancer Research 2020; 2020 Apr 27-28 and Jun 22-24. Philadelphia (PA): AACR; Cancer Res 2020;80(16 Suppl):Abstract nr 4044.
Abstract 4064: Optimization of the transposon expression system for electroporation based gene delivery
Authors: Chen Jian , Chen Youbang 
Published: September 2020

Abstract
Stable protein expression in therapeutic cells such as T cells and NK cells is important for long-lasting clinical effects in cell therapy such as CAR-T therapy. Stable protein expression can be achieved by the viral vector expression method, transposon based expression method and CRISPR/Cas9 mediated knock-in method. Transposon based expression method has the advantage of being relatively safe and cost-effective compared with other methods. However, in the actual application of the transposon systems such as the piggyBac (PB) system and the Sleeping Beauty (SB) system, it is quite challenging to achieve a high delivery efficiency and a high cell viability. We found that in addition to the choice of electroporation methodology, vector design and cell culture are two most important aspects that need to be optimized. Vector design dictates protein expression level as commonly anticipated, it also dictates the expression timing. We found that two distinct designs give two very different expression timing, one with initial high efficiency of expression which tapers down to about half, and another one with very low initial expression level and it goes up to a very high level after two weeks. We propose that the differential expression profiles of the vector systems can be applied to different circumstances to achieve better therapeutic effects.
Citation Format: Jian Chen, Youbang Chen. Optimization of the transposon expression system for electroporation based gene delivery [abstract]. In: Proceedings of the Annual Meeting of the American Association for Cancer Research 2020; 2020 Apr 27-28 and Jun 22-24. Philadelphia (PA): AACR; Cancer Res 2020;80(16 Suppl):Abstract nr 4064.
Abstract 4072: Analysis of clinical BRCA1/2 reversions identifies hotspot mutations and predicted neoantigens associated with therapy resistance
Authors: Pettitt Stephen , Frankum Jessica , Punta Marco , Lise Stefano , Alexander John , Chen Yi , Haider Syed , Tutt Andrew N. , Lord Christopher J. 
Published: September 2020

Abstract
Although PARP inhibitors and platinum salts form part of the standard-of-care for cancers with homologous recombination (HR) defects, drug resistance does emerge and is often caused by reversion mutations in BRCA1 or BRCA2. Reversion mutations are secondary mutations that restore functional protein or compensate for frameshift mutations. To better understand the nature and aetiology of these mutations, we collated, codified and analysed over 200 reversion mutations from 24 published studies. Most reversions have been reported as case reports or small clinical series - we have collected all published reports into a single freely-accessible database to which further cases can be submitted.
The majority of reported reversion mutations are from BRCA1/2 mutant serous ovarian cancer, reflecting the longer use of platinum and PARP inhibitors in this disease. Our analysis identified reversion “hotspots” and “deserts” in the N- and C-terminal coding regions (respectively) of BRCA2, suggesting that pathogenic mutations in these domains may be at higher or lower chance of reversion, an effect not seen for BRCA1. Missense and splice-site pathogenic mutations in BRCA1/2 also appeared less likely to revert than frameshift-causing mutations. Unexpectedly, whilst most reversions were &lt;100 bp “second site” deletions, microhomology use was not universal, especially in BRCA1-mutant cancers, suggesting that multiple DNA repair processes cause reversion and these vary in BRCA1 vs. BRCA2 mutant tumours. Reversions in BRCA1 mutant tumours were less likely to be mediated by deletions.
Many reversions contain novel protein sequence not found in the wild type protein, for example where a stretch of out-of-frame protein sequence is retained between the pathogenic and reversion mutations. We modelled MHC binding affinities for these sequences and found that many reversions were predicted to encode potentially immunogenic neopeptides, suggesting a route to the treatment of reverted disease. These observations have implications for how drug resistance might be managed in BRCA-mutant cancers.
Citation Format: Stephen Pettitt, Jessica Frankum, Marco Punta, Stefano Lise, John Alexander, Yi Chen, Syed Haider, Andrew N. Tutt, Christopher J. Lord. Analysis of clinical BRCA1/2 reversions identifies hotspot mutations and predicted neoantigens associated with therapy resistance [abstract]. In: Proceedings of the Annual Meeting of the American Association for Cancer Research 2020; 2020 Apr 27-28 and Jun 22-24. Philadelphia (PA): AACR; Cancer Res 2020;80(16 Suppl):Abstract nr 4072.
Abstract 4074: Reversing acquired PARPi resistance of TNBC through combined inhibition of cMet and EGFR
Authors: Chu Yu Yi , Yam Clinton , Chen Mei-Kuang , Chan Li-Chuan , Nie Lei , Hung Mien-Chie 
Published: September 2020

Abstract
Front line therapeutic failure happened to half of triple-negative breast cancer (TNBC) patients represent the effective treatment strategies for these patients are urgently needed. Poly (ADP-ribose) polymerase (PARP) is currently the most promising drug target for BRCA-mutated TNBC, and PARP inhibitors (PARPi), olaparib and talazoparib, were recently approved for the treatment of metastatic breast cancer (including TNBC) in patients with germline BRCA1/2 mutations. Despite impressive response rates of ~60%, the prolongation in median progression-free survival with a PARPi is modest, suggesting the emergence of resistance. Several studies have reported that receptor tyrosine kinases (RTKs), such as c-MET (also known as hepatocyte growth factor receptor), are involved in resistance to various anti-neoplastic agents, including PARPi. However, the mechanism by which c-MET contributes to acquired resistance to PARPi in TNBC is not fully understood. In this study, we analyzed acquired PARPi resistant TNBC cells and found c-Met is hyperactivated in these cells with PARPi treatment. We further treated these cells with talazoparib and crizotinib (a multi-kinase inhibitor that inhibits c-MET) and found synergistic inhibition effects of cell proliferation. Unexpectedly, limited effects of talazoparib sensitivity showed while depleting c-MET in PARPi-resistant cells. Interestingly, we found epidermal growth factor receptor (EGFR) is also hyperactivated and interaction of EGFR/c-Met in these cells. Notably, combination of c-MET and EGFR inhibitors increased sensitivity to talazoparib in TNBC cells with acquired resistance to PARPi. Combining EGFR and PARP inhibitors also resulted in greater inhibition of proliferation in c-MET-depleted TNBC cells. Collectively, our findings suggested a potential combination therapy of inhibiting c-MET and EGFR to overcome acquired PARPi resistance in TNBC.
Citation Format: Yu Yi Chu, Clinton Yam, Mei-Kuang Chen, Li-Chuan Chan, Lei Nie, Mien-Chie Hung. Reversing acquired PARPi resistance of TNBC through combined inhibition of cMet and EGFR [abstract]. In: Proceedings of the Annual Meeting of the American Association for Cancer Research 2020; 2020 Apr 27-28 and Jun 22-24. Philadelphia (PA): AACR; Cancer Res 2020;80(16 Suppl):Abstract nr 4074.
Abstract 4102: Genome-wide CRISPRa screen reveals new drivers of Enzalutamide resistance in prostate cancer
Authors: Rodriguez Yara , Vatapalli Rajita , Han Huyin , Unno Kenji , Sagar Vinay , Chalmers Zachary , Lysy Barbara , Yoo Young A , Truica Mihai , Abdulkadir Sarki 
Published: September 2020

Abstract
Potent anti-androgens such as Enzalutamide (Enza) have been approved for the treatment of mCRPC. Despite initial response, nearly all patients eventually develop resistance to Enza. Although a few mechanisms of Enza resistance have been identified, none have been translated into a clinical benefit for recurrent patients. Our goal is to identify new therapeutic targets to overcome Enza resistance. We performed a genome-wide CRISPR activation screen using deactivated Cas9 (dCas9) fused to VP64 transcriptional activator in the androgen-dependent prostate cancer cell line (LNCaP). We used the Calabrese sgRNA library which targets 18,885 genes using 3 different sgRNAs per gene (56,762 sgRNAs). Cells expressing the gRNA library were treated with Enza (20µM) for 7 weeks to allow for enrichment of potential resistance genes. We identified 10 genes which were targeted by at least 2 sgRNAs with a Fold Change (FC) &gt; 4 in the Enza vs control group. Notably, AR was the most enriched gene. AR up-regulation is one of the main drivers of Enza resistance in patients. Pathway enrichment analysis using all the sgRNAs with FC&gt; 5 revealed an over-representation of the RB1, TP53 and TGF-β pathways. Importantly, these pathways are also known to drive Enza resistance. Our screen also identified new candidate genes that have never been linked to Enza resistance. Interestingly, most of these genes are transcription factors. We validated the screen results in vitro. Further analysis of this target genes in vitro and in vivo will allow us to identify novel genes and potential therapeutic targets for Enza resistance.
Citation Format: Yara Rodriguez, Rajita Vatapalli, Huyin Han, Kenji Unno, Vinay Sagar, Zachary Chalmers, Barbara Lysy, Young A Yoo, Mihai Truica, Sarki Abdulkadir. Genome-wide CRISPRa screen reveals new drivers of Enzalutamide resistance in prostate cancer [abstract]. In: Proceedings of the Annual Meeting of the American Association for Cancer Research 2020; 2020 Apr 27-28 and Jun 22-24. Philadelphia (PA): AACR; Cancer Res 2020;80(16 Suppl):Abstract nr 4102.