bluuebunny/crossref_metadata_2025_split huggingface dataset:
Abstract 4514: Discovery of potent and orally bioavailable degraders of HPK1 based on a novel HPK1 binder
Authors: Zhang Zhimin , Wu Mengqiang , Wang Ling , Yang Xi , Zhang Zhiping , Guo Liubin , Pan Hao , Zhao Mengting , Wang Linli , Liu Sirui , Dong Zhao , Jiang Chunhua , Zheng Haowen , Liu Dongzhou 
Published: March 2024

Abstract
Background: Proteolysis targeting chimeras (PROTACs) are novel structures designed to target a protein of interest (POI) for ubiquitination and degradation, leading to the selective reduction in the expression levels of the POI. Hematopoietic progenitor kinase 1 (HPK1) is considered a promising target for tumor immunotherapy. PROTAC-involved disruption of HPK1 protein activity enhanced the treatment efficacy of CAR-T-cell-based immunotherapies in different solid tumors and mouse models. Thus, targeting HPK1 by PROTAC to restore T-cell activity is an attractive approach to induce a greater immune responses in cancer.
Results: We designed and synthesized a series of HPK1 degraders by tethering our previously discovered novel HPK1 inhibitor with CRBN binder through various linkage. The most potent PROTAC effectively degrades HPK1 proteins in the Ramous cell in a dose-dependent manner and achieves DC50 value &lt;50 nM. The maximum degradation (D max ) values achieved are &gt;90%. Our PROTAC showed EC50 value &lt;100 nM for IL-2 and IFNγ production. PK studies revealed a low clearance, high plasma exposure and oral bioavailability in mice and rats. Furthermore, our PROTAC is shown to induce robust and statistically significant tumor growth inhibition in the MC38 and CT26 syngeneic model with increased T cell signatures observed within the tumor. The PROTAC in combination with anti-PD1 also resulted in robust anti-tumor activity with TGI &gt; 90%. The 14 days DRF experiment shows that the compound has a great safety window(&gt;200 folds).
Conclusion: We show here that a potent oral HPK1 degrader demonstrates strong immune cell activation and robust anti-tumor activity in mouse syngeneic tumor models, as a single agent and in combination with anti-PD1. Further evaluation of these potent degraders in additional in vivo studies will continue to build upon our mechanistic understanding of HPK1 degradation as a novel immunomodulatory approach for anti-tumor immunity.
Citation Format: Zhimin Zhang, Mengqiang Wu, Ling Wang, Xi Yang, Zhiping Zhang, Liubin Guo, Hao Pan, Mengting Zhao, Linli Wang, Sirui Liu, Zhao Dong, Chunhua Jiang, Haowen Zheng, Dongzhou Liu. Discovery of potent and orally bioavailable degraders of HPK1 based on a novel HPK1 binder [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 4514.
Abstract 455: Identification of hsa-miR-6794-3p as a key regulatory factor in human pancreatic cancer metastasis
Authors: Kim Ha Gyeong , Oh Eun-Taex , Cho Yunmi , Park Heon Joo 
Published: March 2024

Abstract
Metastasis is a major factor in treatment failure in patients with pancreatic cancer, but the underlying mechanisms remain unclear, and new, effective treatment strategies are needed. Here, we sought to identify novel microRNAs (miRNAs) with a key role in pancreatic cancer metastasis. Using a microarray analysis to evaluate miRNA expression in metastatic and non-metastatic pancreatic cancer, we found that expression of the miRNA, hsa-miR-6794-3p, was significantly lower in metastatic pancreatic cancer tissues than in non-metastatic pancreatic cancer tissues. Methylation-specific polymerase chain reaction analyses revealed that hsa-miR-6794-3p expression was downregulated in metastatic pancreatic cancer cells by hypermethylation of CpG islands within the hsa-miR-6794-3p promoter. Gain- and loss-of-function approaches using the human pancreatic cancer cell lines, MIA-PaCa-2 and HPAF-II, expressing low and high levels of hsa-miR-6794-3p, respectively, suggested a role for hsa-miR-6794-3p in decreasing cell invasion, migration, and epithelial-mesenchymal transition (EMT) signaling. Notably, we found that hsa-miR-6794-3p exerts its effects by inhibiting expression of the chromatin remodeling factor, RBBP4, through direct binding to the 3’-untranslated region of its mRNA. The resulting suppression of RBBP4 expression increases levels of the transcription factor GRHL2, which is involved in regulating invasion, migration, and EMT signaling in metastatic pancreatic cancer cells. Thus, lower levels of hsa-miR-6794-3p disrupt GRHL2-mediated regulation of these pathways by disinhibiting expression of the GRHL2 negative regulator, RBBP4. Consistent with these findings, an investigation of the relationship between hsa-miR-6794-3p and clinicopathological features in pancreatic cancer patients using publicly available datasets and immunohistochemistry showed that low levels of hsa-miR-6794-3p are correlated with a poorer prognosis in pancreatic cancer patients. Additional preclinical experiments in nude mice clearly demonstrated that hsa-miR-6794-3p is capable of suppressing pancreatic cancer cell metastasis. Taken together, these results suggest that hsa-miR-6794-3p suppresses tumor metastasis and thus represents a promising prognostic biomarker and therapeutic target in pancreatic cancer.
Citation Format: Ha Gyeong Kim, Eun-Taex Oh, Yunmi Cho, Heon Joo Park. Identification of hsa-miR-6794-3p as a key regulatory factor in human pancreatic cancer metastasis [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 455.
Abstract 4567: Dual targeting of HSP70 and AURKA improves treatment in neuroendocrine prostate cancer
Authors: Xu Pengfei , Yang Joy C. , Chen Bo , Ning Shu , Wang Leyi , Nip Christopher , Evans Christopher P. , Dall'Era Marc A. , Gao Allen C. , Gestwicki Jason , Liu Chengfei 
Published: March 2024

Abstract
Background: Neuroendocrine prostate cancer (NEPC) is the most aggressive type of prostate cancer with no effective treatments. Therefore, there is an urgent need to develop new treatment strategies. N-Myc plays a key role in driving NEPC progression, and Aurora kinase A (AURKA) prevents N-Myc degradation by interact with N-Myc and disallowing the intervention of ubiquitin ligases. The AURKA inhibitor Alisertib inhibits NEPC tumor growth by disrupting N-Myc signaling, but it is failed in a phase II clinical trial. In this study, we tested the synergy of novel HSP70 allosteric inhibitor JG231 and Alisertib in NEPC cells, organoids, and patient derived xenograft (PDX) models, which will provide new therapeutic strategies for the NEPC treatment.
Methods: RT-PCR, western blotting, or immunohistochemistry (IHC) were used to determine the expression of N-Myc, neuroendocrine maker (NSE, SYP, CHGA, etc) in different prostate cancer cell lines and PDX tumors. After the knockdown of HSP70 and AURKA by siRNA transfection, cell proliferation and N-Myc expression were assessed by using cell viability assay, RT-PCR and Western blotting respectively. The co-immunoprecipitation assay was performed to detect the N-Myc ubiquitination. RNA-sequencing, gene set enrichment analysis was employed to determine the changes of JG231 on gene programs. The effects of JG231 and Alisertib on the proliferation were examined in NEPC cell lines and PDX organoids models.
Results: We determined that PC3, CWR22Rv1, H660 cell lines, LuCaP49, and LuCaP93 PDX tumors had significantly increased the expression of N-Myc and neuroendocrine makers. Knockdown of HSP70 or using the HSP70 inhibitor, JG231, significantly inhibit the growth of H660 and CWR22Rv1 cells and leads to a synergistic effect with Alisertib (p&lt;0.001). JG231 combined with Alisertib also inhibits growth and induces the death of LuCaP93 and H660 tumor derived organoids in a dose-dependent manner (p&lt;0.001). Mechanistically, JG231 inhibits the protein expression of N-Myc through the ubiquitin-proteasome system and promote STUB1 to enter the nucleus to bind to N-Myc.
Conclusions: JG231, a novel HSP70 inhibitor, can improves the treatment efficacy of AURKA inhibitor Alisertib against NEPC through the regulation of N-Myc protein homeostasis.
Citation Format: Pengfei Xu, Joy C. Yang, Bo Chen, Shu Ning, Leyi Wang, Christopher Nip, Christopher P. Evans, Marc A. Dall'Era, Allen C. Gao, Jason Gestwicki, Chengfei Liu. Dual targeting of HSP70 and AURKA improves treatment in neuroendocrine prostate 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 4567.
Abstract 4620: Exploring the role of GHRH antagonist MIA-602 in overcoming doxorubicin resistance in acute myeloid leukemia
Authors: Gaumond Simonetta I. , Costoya Joel , Schally Andrew V. , Jimenez Joaquin J. 
Published: March 2024

Abstract
Acute Myeloid Leukemia (AML) is characterized by the uncontrolled proliferation of immature hematopoietic progenitors, often associated with a poor prognosis. Conventional therapies involve chemotherapy and bone marrow stem cell transplantation. The growth hormone releasing hormone receptor (GHRH-R) antagonist MIA-602 has exhibited inhibitory effects on the growth of various human cancer cell lines, including AML cells. This study aimed to investigate the impact of MIA-602 alone and in combination with doxorubicin on three doxorubicin-resistant AML cell lines, K-562, U-937, and KG-1A. Initial examinations confirmed the presence of GHRH receptors in both wild-type and doxorubicin-resistant cell lines through western blot analysis. Doxorubicin was administered at concentrations of 0.005, 0.01, and 0.05 μg/ml, while MIA-602 was used at 5 μmol/L. Subsequent experiments involved culturing both wild-type and doxorubicin-resistant clones with MIA-602 at concentrations ranging from 0.05 μmol/L to 5 μmol/L for 24 and 48 hours. The results revealed a significant, dose- and time-dependent reduction in cell proliferation across all six cell lines. Both wild-type and doxorubicin-resistant clones exhibited a comparable decrease in cell proliferation when exposed to MIA-602 at concentrations greater than 0.05 μmol/L (p &lt; 0.05) after 24 and 48 hours. In conclusion, our study demonstrates that these three AML cell lines and their doxorubicin-resistant clones remain susceptible to GHRH antagonist MIA-602. These findings may pave the way for the development of novel therapies or drug combinations using GHRH antagonists such as MIA-602 for treating AML. This is of particular interest for those who are resistant to conventional chemotherapy, further broadening the spectrum of treatment options available. Additional investigations are warranted to progress these findings to in vivo xenograft models.
Citation Format: Simonetta I. Gaumond, Joel Costoya, Andrew V. Schally, Joaquin J. Jimenez. Exploring the role of GHRH antagonist MIA-602 in overcoming doxorubicin resistance in acute myeloid leukemia [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 4620.
Abstract 4669: Differential preclinical activity of Bruton tyrosine kinase inhibitors (BTKis) against BTK resistance-associated mutations
Authors: Davids Matthew S. , Ahn Inhye E. , Riehm Jacob J. , Acar Melih , Girardi Vincent , Patel Sima , Gururaja Tarikere , Woyach Jennifer , Lam Lloyd T. , Szafer-Glusman Edith 
Published: March 2024

Abstract
Background: Differences in profiles of acquired mutations of resistance to BTKis might have implications for treatment sequencing. We evaluated preclinical activity of BTKis (ibrutinib [Ibr], acalabrutinib [Acala], zanubrutinib [Zanu], and pirtobrutinib [Pirto]) against multiple clinically relevant BTK mutations and report on the clinical impact of BTK C481S in patients (pts) treated with Ibr.
Methods: Inhibition of kinase activity by BTKis was evaluated in a cell-free kinase activity assay (Nanosyn). Cell-killing activity on CRISPR-generated BTK mutant knock-in TMD8 cells was assessed after 3 days of BTKi treatment. Time from first detection of C481S mutation to progressive disease (PD) was evaluated in pts treated with Ibr with previously untreated (n=247) or relapsed/refractory (R/R; n=172) CLL using pooled data from 5 clinical trials.
Results: In vitro kinase activity was highest for Ibr compared with Acala or Zanu against C481S (Ibr IC50: 26.3 nM; Zanu: 84.8 nM; Acala: 338 nM) and T474I (Ibr: 1.2 nM; Zanu: 4.4 nM; Acala: 43.6 nM). Ibr maintained cell-killing activity against all tested BTK mutants, whereas limited activity was seen with Acala against C481S and A428D; with Zanu against C481S, L528W, and A428D; and with Pirto against T474I, L528W, V416L, and A428D (Table). In pts treated with Ibr, C481S was detected prior to clinical PD in 3 previously untreated pts and in 26 pts with R/R CLL; median (range) time from first detection of C481S to PD in these pts was 17.0 mo (11.2-34.3) and 10.0 mo (2.6-35.7), respectively.
Conclusion: This study demonstrates activity of Ibr against all evaluated BTK variants, which are associated with resistance to covalent and noncovalent BTKis. Clinical data shows that pts who develop C481S mutations continue to derive benefit from Ibr for several months before PD. Together, these data suggest a role for Ibr in the treatment of pts with a broad spectrum of BTK variants.
Table. Cell-killing activity of BTKis against wild type and mutant BTK EC50 (nM) BTK Wild type C481S T474I L528W V416L A428D Ibr 0.6 414.0 1.9 181.4 0.5 205.3 Acala 3.9 2973.8 43.3 3.5 106.3 3000.0 Zanu 0.7 2322.0 11.7 3000.0 0.6 1726.0 Pirto 8.2 12.6 2727.5 3000.0 1898.0 3000.0
Citation Format: Matthew S. Davids, Inhye E. Ahn, Jacob J. Riehm, Melih Acar, Vincent Girardi, Sima Patel, Tarikere Gururaja, Jennifer Woyach, Lloyd T. Lam, Edith Szafer-Glusman. Differential preclinical activity of Bruton tyrosine kinase inhibitors (BTKis) against BTK resistance-associated mutations [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 4669.
Abstract 4703: Automated 42 PDX 3D in vitro tumor models of the TME screen immuno-oncology and targeted compounds and biologics for antitumor effects and MOA
Authors: Xue Bin , Harrod Christopher , Le Long , Schüler Julia , Hribar Kolin C. 
Published: March 2024

Abstract
High throughput screening has been widely utilized for its efficiency in testing novel therapeutic agents. However, the conventional screening using 2D culture of cancer cell lines lacks the biological complexity of the tumor microenvironment or the immune compartment which limits their application in testing immune-oncology therapies. Particularly with the passing of the FDA Modernization Act 2.0, it is critical to create more biologically relevant 3D in vitro models that can be used to reduce, refine and ultimately replace preclinical animal models. Here we have developed a highly automated 3D high throughput screening platform comprised of 42 patient-derived xenograft models in coculture with fibroblasts in engineered extracellular matrix hydrogels that resemble tumor stromal microenvironment. The PDX models cover a diverse range of tumor and histological types including NSCLC, CRC, gastric, TNBC, RCC, bladder, ovarian, melanoma, pancreatic, and others. Moreover, PBMCs or other immune cells can be incorporated into the system for evaluating immuno-oncology drugs. The panel has been tested against chemotherapy drug cisplatin and the immunotherapy biologic, Solitomab, an EpCAM/CD3 bispecific antibody. The endpoint analyses for antitumor effects include the dose response of tumor size and tumor cell death percentage based on high content imaging analysis. In addition, immune cells and supernatant in the IO assay can be recovered from the wells and used for flow cytometry profiling and cytokine assays. In summary, the novel 42-PDX panel described here offers a unique and powerful tool for rapidly generating preclinical data and a better understanding the drug activity at the pharmacology stages, opening the door for faster and more human relevant drug discovery for cancer patients.
Citation Format: Bin Xue, Christopher Harrod, Long Le, Julia Schüler, Kolin C. Hribar. Automated 42 PDX 3D in vitro tumor models of the TME screen immuno-oncology and targeted compounds and biologics for antitumor effects and MOA [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 4703.
Abstract 472: Long non-coding RNA MIR31HG-derived circRNA enhances the oncogenicity of oral carcinoma
Authors: Chou Chung-Hsien , Tu Hsi-Feng , Chang Kuo-Wei , Lin Shu-Chun 
Published: March 2024

Abstract
Introduction: Oral carcinoma (OC) represents a significant health challenge, with the role of non-coding RNAs, particularly microRNAs and long non-coding RNAs (LncRNAs), being well-established in its progression. However, the implications of circular RNAs (circRNAs) in this context are less well understood, presenting a notable gap in current research. Our study focuses on the MIR31HG LncRNA, identified as crucial in OC, particularly its role in producing circRNA, as evidenced by sequencing. This specific MIR31HG-derived circRNA, designated 31HGcircR, which contains a potential open reading frame (ORF), and may demonstrate phenotypic significances. Alterations to 31HGcircR, either through knockout or ORF mutation, result in remarkable phenotypic changes, highlighting its potential importance in the pathogenesis of OC.
Methods: We implemented 31HGcircR overexpression in OC cell lines, utilizing the TetOff system for gene expression control. CRISPR/Cas9 was employed for gene knockout, and siRNA for gene knockdown. Gene expression was quantified using qPCR. Additionally, the BaseScope assay was used to label and detect the spatial expression of overexpressed 31HGcircR within the cells. Assays to assess cell proliferation, invasion, migration, anchorage-independent growth (AIG), and clonogenicity were conducted.
Results: The findings revealed specific junctions of 31HGcircR and varied expression levels across different OC cell lines. There was a significant correlation among the expression of 31HGcircR, MIR31HG and miR-31-5p. Notably, the knockdown of 31HGcircR led to pronounced inhibition of cell growth, migration, invasion, AIG, and clonogenic activity. Conversely, overexpression of 31HGcircR or its ORFs augmented invasion and clonogenic potential in certain cell lines. The TetOff system demonstrated that minimal concentrations of Doxycycline effectively suppressed 31HGcircR expression, with reversible effects upon removal. Knocking out 31HGcircR resulted in reduced invasion and clonogenic capabilities.
Conclusion: This study provides clues demonstrating the oncogenic roles of 31HGcircR in OC, which simulate the functions of MIR31HG or miR-31-5p. The changes in cell phenotypes following 31HGcircR manipulation suggest its potential as a therapeutic target or biomarker in OC. These findings offer valuable insights, contributing to the evolving understanding of circRNAs in cancer research and potential treatment avenues.
Citation Format: Chung-Hsien Chou, Hsi-Feng Tu, Kuo-Wei Chang, Shu-Chun Lin. Long non-coding RNA MIR31HG-derived circRNA enhances the oncogenicity of oral 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 472.
Abstract 4778: Development of self-amplifying RNA vaccines targeting prostate tissue-restricted and tumor-associated antigens
Authors: Shoemaker Robert H. , Erasmus Jesse H. , Berglund Peter , Marshall Jason D. , Koboziev Yuri , Beckman Bradley M. , Cholewa Brian D. , Reed Steven G. 
Published: March 2024

Abstract
Prostate cancer is the most frequent cancer type in men, with increasing incidence at advanced age. As this cancer is frequently slow growing, opportunities for cancer interception exist, particularly among men with low-grade tumors who frequently undergo active surveillance. A well-tolerated vaccine intervention would be ideal for this population. Self-amplifying RNA (saRNA) vaccines built on a replication-deficient alphavirus backbone such as VEEV, offer several advantages for vaccine development. Sequences for relatively large antigens can be encoded and efficiently expressed, at low vaccine doses. Practical and efficient delivery of the saRNA is possible with advanced nanoparticles such as cationic inorganic lipid nanoparticles (LION). Among candidate antigens for a prostate cancer interception vaccine, we selected prostein (SLC45A3), a protein whose natural expression is restricted to the prostate gland and prostate cancer and ERG, a member of the ETS family of transcription factors frequently involved in chromosomal translocations in prostate cancer. We generated saRNA constructs with several design strategies and demonstrated that upon in vitro transfection, both antigens are expressed as assessed by mass spectrometry and Western blot analysis. This was achieved using commercial transfection reagents and LION. Prostein expression was particularly notable in that this eleven-transmembrane protein could be detected in HEK-293 cells using flow-cytometry and cell-surface staining. Nearly 50% of the cell population stained positively. Immunogenicity of the vaccines was tested in mice. Following formulation with LION particles i.m. injection and boosting, saRNAs coding for prostein elicited strong cellular immunity as demonstrated by IFN-γ ELISPOT analysis. Detailed analysis of intracellular cytokine expression indicated a strong CD8 response. saRNAs coding for ERG also showed antigen-specific ELISPOTs. Using commercially available recombinant ERG protein, an ELISA was developed and a strong humoral response to the ERG saRNA vaccine was demonstrated. The saRNA vaccines will be further credentialled with additional preclinical studies including cancer preventive efficacy studies in genetically engineered mouse models of prostate cancer.
Citation Format: Robert H. Shoemaker, Jesse H. Erasmus, Peter Berglund, Jason D. Marshall, Yuri Koboziev, Bradley M. Beckman, Brian D. Cholewa, Steven G. Reed. Development of self-amplifying RNA vaccines targeting prostate tissue-restricted and tumor-associated antigens [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 4778.