tbuckley/cabot-search huggingface dataset:
Abstract 630: The role of AKT pathway in β-catenin mutated hepatocellular carcinoma
Authors: Xue Wang, Xin Chen
Journal: Cancer Research (2024)

Abstract The Wnt/β-catenin and PI3K/AKT pathways are pivotal contributors to the development of the malignant phenotype in cancer. The activated Wnt/β-catenin pathway synergizes with various signaling cascades to propel HCC formation, operating through its downstream effectors. It has been well-established that the mammalian target of rapamycin complex 2 (mTORC2) serves as an upstream regulator of AKT, orchestrating the phosphorylation and activation of AKT. Once activated, AKT phosphorylates and inhibits canonical downstream targets like forkhead box O protein (FOXO) and tuberous sclerosis complex 2 (TSC2). Studies indicated that AKT-mediated phosphorylation of TSC2 activates mTORC1, subsequently suppressing autophagy. However, there are limited data exist on the interplay between Wnt/β-catenin and AKT pathways in HCC. In this study, we sought to explore the role of the AKT pathway in β-catenin activated HCC tumors. Sleeping beauty-mediated hydrodynamic transfection was used to overexpress constitutively active forms of β-catenin (N90-β-catenin) and c-MET in the livers of mice to establish the HCC model (c-Met/β-catenin model). Notably, the knockout of Rictor, a component of the mTORC2 complex, significantly impeded tumor growth, underscoring the pivotal role of AKT in c-Met/β-catenin tumor development. AKT1 and AKT2 are the two main isoforms expressed in the liver. We determined that both AKT1 and AKT2 isoforms are critical for c-Met/β-catenin tumor growth. Intriguingly, simultaneous knockout of Rictor and activation of mTORC1 in the c-Met/β-catenin model resulted in delayed tumor growth, indicating that mTORC1 is not the sole downstream determinant of AKT signaling during c-Met/β-Catenin tumor development. Furthermore, overexpression of the activated FOXO1 (FOXO1AAA) significantly delayed the tumor growth, which highlighted the importance of FOXO1. Given the limited understanding of FOXO1 in HCC, we extended our investigation on FOXO1 to human HCC cell lines. Overexpression of FOXO1AAA notably delayed tumor growth in vitro. RNAseq analysis of FOXO1AAA-overexpressed human HCC cell lines revealed a significant upregulation of "metabolic pathways", suggesting metabolic reprogramming post FOXO1 activation. This study, for the first time, unveils the role of the AKT pathway in β-catenin-mutated HCC growth and highlights the potential therapeutic value of targeting the AKT pathway in patients with β-catenin-mutated HCC. Citation Format: Xue Wang, Xin Chen. The role of AKT pathway in β-catenin mutated hepatocellular 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 630.
Abstract 2339: GPNotebook: A toolkit for identifying cancer-related glycosylation alterations in glycoproteomic analysis
Authors: Hui Zhang, Trung Hoang, Yingwei Hu
Journal: Cancer Research (2024)

Abstract This study is dedicated to the development and application of 'GPNotebook', an innovative Python-based data analysis tool, specifically designed to enhance glycopeptide research in cancer studies by enabling the detailed investigation of glycosylation patterns and their correlation with various clinical cancer phenotypes. In our study, we recognize the critical role of protein glycosylation in biological processes and its relevance in cancer. To address the lack of specialized tools for glycoproteomic analysis, we developed GPNotebook, a comprehensive Python package designed for detailed investigation of intact glycopeptides (IGPs). This tool integrates multiple functionalities, including statistical profiling, differential expression analysis, categorization of glycosylation subtypes, and exploration of the interactions between glycosylation and phosphorylation. It also encompasses survival analysis and assessments of glycosylation enzymes. We applied GPNotebook in a research project focused on 10 cancer types collected from Clinical Proteomic Tumor Analysis Consortium (CPTAC) projects to validate its efficacy and demonstrate its wide-ranging capabilities. Through this application, we were able to analyze and interpret complex glycoproteomic data, revealing potential IGPs as biomarkers for cancer detection and differentiation of cancer subtypes. We identified hundreds of cancer-specific IGPs, with several showing promise as early-stage cancer biomarkers. In addition, our analysis revealed unique glycosylation patterns linked to specific molecular subtypes and immune cell distributions in tumor microenvironments. We also clarified the roles of certain glycosylation enzymes across different cancer types, highlighting their potential significance in cancer progression. GPNotebook has proven to be a critical tool in glycopeptide research. Our findings not only enhance the understanding of glycosylation in cancer but also demonstrate GPNotebook's potential in identifying novel biomarkers and providing insights into cancer subtype differentiation. This study positions GPNotebook as a valuable resource for exploring the complex relationships between protein glycosylation and cancer phenotypes. Citation Format: Hui Zhang, Trung Hoang, Yingwei Hu. GPNotebook: A toolkit for identifying cancer-related glycosylation alterations in glycoproteomic analysis [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 2339.
Abstract 1572: <i>POU5F1B</i> is a human-restricted ROCK inhibitor-sensitive oncoprotein that restructures membrane nanodomains to increase cell adhesion
Authors: Laia Simó-Riudalbas, Sandra Offner, Laurence Abrami, Eduard M. Unterauer, Julien Duc, Evarist Planet, Ralf Jungmann, Gisou van der Goot, Didier Trono
Journal: Cancer Research (2024)

Abstract Evolution confers new species with distinctive biological features, which can translate in purely mechanistic speciation or in novel phenotypic traits. POU5F1B arose by retrotransposition of POU5F1 (OCT4) in the last common ancestor of modern Hominoidae. Its human product is endowed with oncogenic properties, notably promoting gastrointestinal cancer growth and metastasis. Here, we reveal that the oncogenic action of POU5F1B requires ubiquitination of lysine residues found only in the human protein. This post-translational modification is essential for POU5F1B to localize to the cytoplasm and become acylated by the DHHC17 palmitoyltransferase. This leads to POU5F1B association with detergent-resistant membrane subdomains, where it triggers the accumulation of integrins and signaling molecules, and to a stimulation of cell focal adhesion. Finally, we determined that POU5F1B stability is critically dependent on ROCK, a crucial regulator of cytoskeletal dynamics and cell motility. Citation Format: Laia Simó-Riudalbas, Sandra Offner, Laurence Abrami, Eduard M. Unterauer, Julien Duc, Evarist Planet, Ralf Jungmann, Gisou van der Goot, Didier Trono. POU5F1B is a human-restricted ROCK inhibitor-sensitive oncoprotein that restructures membrane nanodomains to increase cell adhesion [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 1572.
Abstract 3451: Association of multi-level factors with cervical cancer screening in healthcare settings
Authors: Rutu Rathod, Ellen Hu, Sandi Pruitt, Jasmin A. Tiro, Jessica Chubak, Jennifer Haas, Steven J. Atlas, Gina Kruse, Amy E. Hughes
Journal: Cancer Research (2024)

Abstract Background- Few studies have examined the independent contribution of patient, clinician and system factors associated with completing cervical cancer screening. Methods- Using a large retrospective cohort created from electronic health record data, we examined the association of patient- and provider-level characteristics with cervical cancer screening among unknown- and average-risk, screen eligible women who entered the cohort at a primary care visit in 1 of 3 U.S. healthcare systems (Kaiser Permanente Washington, Parkland Health, Mass General Brigham) between 2010-2019. The outcome was time, in days, from cohort entry to screening. We fit piecewise exponential (PWE) survival models, nesting patients hierarchically within provider seen and the facility at the time of their analytic cohort entry visit. We excluded patients whose visit was with providers at their non-principal facility (principal defined as the facility that they saw most patients in 2010). The analysis was stratified by healthcare setting. Results- Median time from cohort entry until screening varied across settings (202-1063 days) and across providers and clinics (between cluster variance= 0.11-0.77, 0.06-1.18, respectively). Many patient-level characteristics (age, cohort entry year, race/ethnicity, number of comorbidities, smoking status, primary care visit within 12 months before cohort entry, and insurance), some of the provider-level characteristics (provider specialty and sex) and social determinants of health measures (Local Isolation Score Hispanic) were associated with time to screening. In the adjusted PWE models, for all three settings: a) Younger patients (21-49 years old) (vs older patients (50-65 years)); b) Hispanic and Non-Hispanic Asian/Pacific Islander (vs Non-Hispanic White); and c) patients seeing ob-gyn providers (vs family medicine providers) had shorter time to screening. Patients with a) higher (vs lower) comorbidity burden, and b) Medicare insurance (vs Commercial) were less likely to receive screened. Conclusion- By identifying characteristics at multiple levels that are associated with the outcome, our results help us identify targets for interventions to improve and increase timely cervical cancer screening in healthcare settings. Citation Format: Rutu Rathod, Ellen Hu, Sandi Pruitt, Jasmin A. Tiro, Jessica Chubak, Jennifer Haas, Steven J. Atlas, Gina Kruse, Amy E. Hughes. Association of multi-level factors with cervical cancer screening in healthcare settings [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 3451.
Abstract 4370: Functional characterisation of a novel mutation in PRKCA, a major driver of chordoid gliomas
Authors: Charlotte Bellamy, Hannah Tovell, Florent Dingli, Damarys Loew, Stephane Liva, Selina Schwaighofer, Eduard Stefan, Alexandra Newton, Marc Sanson, Franck Bielle
Journal: Cancer Research (2024)

Abstract Chordoid gliomas (ChG) are a rare low-grade brain tumor, believed to be derived from tanycytes. An analysis by the team identified a novel mutation present in all ChGs: PRKCA p.D463H. This mutation involves a D463H substitution at the kinase domain of the Protein kinase C alpha (PKCα), it is not found in any other cancer, and represents the hallmark of ChG. The aim of the project is to identify novel and biologically relevant PKCαD463H signaling pathways, which will demonstrate the involvement and the role of this mutated kinase in cellular functions implicated in the development of ChGs. The D463H mutation affects a critical residue of the kinase domain of PKCα, suggesting that such a change modifies substrate affinity. Following the purification of PKCαD463H, we have shown its inactivation via in vitro kinase assays and peptide array. In cellulo fret-based activity reporter assays have shown that PKCαD463H has a dominant negative effect over PKCαWT. Through co-immunoprecipitation we have shown that the mutant protein can bind the WT form and also colocalizes in the cell. Our results also show that the mutation affects the tertiary structure of the protein, resulting in a more open, unstable protein compared to the WT. Phosphoproteomic analysis of HEK cells overexpressing PKC⍺D463H show a decrease of phosphorylation specifically on proteins involved with cell-cell adhesion, including functionally relevant phosphosites. This is in line with our Co-IP mass spectrometry data which shows a decrease in interaction of PKC⍺D463H with proteins involved in cell junctions. The cell of origin, tanycytes, are highly specialized cells that contain important cell-cell junctions and therefore their perturbation could be a potential route of tumorigenesis. By understanding these changes integrated with our snRNAseq and bulk RNAseq of ChGs, and exploration in cellular models, we hope to elucidate the mechanism by which the mutation leads to the development of Chordoid Glioma. Citation Format: Charlotte Bellamy, Hannah Tovell, Florent Dingli, Damarys Loew, Stephane Liva, Selina Schwaighofer, Eduard Stefan, Alexandra Newton, Marc Sanson, Franck Bielle. Functional characterisation of a novel mutation in PRKCA, a major driver of chordoid gliomas [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 4370.
Abstract 2029: Investigation of anti-tumor activity of porcine myeloperoxidase for urothelial carcinoma of the bladder
Authors: Junnan Liu, Robert C. Allen, Jackson T. Stephens, Paras Shah, Haojie Huang
Journal: Cancer Research (2024)

Abstract Introduction: Porcine myeloperoxidase (pMPO) is a cationic enzyme secreted by granulocytic leukocytes capable of selectively binding cancer cells due to their negative surface charge. In the presence of hydrogen peroxide (H2O2) and sodium chloride (NaCl), MPO catalyzes the production of electronically excited singlet molecular oxygen, which exerts focused cytotoxic activity against cancer cells. Methods: Human bladder non-cancer (SV-HUC1) and cancer cells (T24 and 5637) were used to assess differential cytotoxic activity of pMPO. pMPO was combined with associated reagents, including H2O2, NaCl, and L-proline and glycine for 45 minutes to support the generation of singlet oxygen. MTS assay was used to assessed cell viability at day 3 post-treatment. Non-cancer and cancer cells were labeled with GFP and mCherry, respectively, via infection with lentivirus and cultured in tandem to allow for simultaneous treatment of both cell types. After the treatment, flow cytometry and microscopy were used to monitor the growth of the cells. Results: No cytotoxicity was noted when SV-HUC1 cells were cultured with individual reagents pMPO, NaCl, L-Proline, or Glycine, and limited cytotoxicity was observed with H2O2. Notably, cell viability was nearly maximally preserved for SV-HUC1 cells. The combination of pMPO with associated reagents resulted in killing of 5637 and T24 cells, with tumoricidal activity maintained across increasing tumor cell density, with cell viability preserved for SV-HUC1 cells. Within the cell co-culture system, simultaneous treatment of both cell types with the pMPO treatment resulted in selective killing of tumor cells and preservation of SV-HUC1 cells. Mechanistic studies confirm specificity of pMPO binding to tumor cells, and further, high levels of expression of gammaH2AX and phospho-ATM in 5637 and T24 cells following treatment. Conclusion: pMPO demonstrates selective and efficient killing of bladder cancer cells with minimal cytotoxicity against non-cancer urothelial cells. Given its reliance on direct surface contact with tumor cells for activity, pMPO may be developed as an intravesical therapy for treatment of non-muscle invasive bladder cancers. Acknowledgements: This work was partially supported by Mayo funds provided to Dr. Haojie Huang. Citation Format: Junnan Liu, Robert C. Allen, Jackson T. Stephens, Paras Shah, Haojie Huang. Investigation of anti-tumor activity of porcine myeloperoxidase for urothelial carcinoma of the bladder [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 2029.
Abstract 1925: Wild-type RAS signaling is an essential therapeutic target in <i>RAS</i>-mutated cancers
Authors: Nancy E. Sealover, Bridget Finniff, Jacob Hughes, Hyun Lee, Robert L. Kortum
Journal: Cancer Research (2024)

Abstract Single-agent targeting of RAS-mutated cancers using RAF/MEK/ERK or PI3K/AKT pathway inhibitors is ineffective; blocking one pathway relieves negative feedback control of RTK signaling and hyperactivates parallel effector pathways. While combined MEK and PI3K inhibition is effective in preclinical models, toxicity of this combination prevents its clinical use. Alternative strategies for treating RAS-mutated cancers are essential. Therapeutics directly targeting mutated RAS proteins have the potential to spare normal cellular function thereby lessening overall toxicity. Unfortunately, direct RAS inhibition as a monotherapy does not fully block RAS effector signaling. RAS proteins show differential activation of RAF and PI3K pathways: KRAS potently activates RAF but poorly activates PI3K, whereas HRAS effectively activates PI3K but poorly activates RAF. Further, mutant RAS inhibition relieves negative feedback controls leading to rapid hyperactivation of RTK−WT RAS signaling. A more comprehensive understanding of the interplay between mutant RAS and RTK−WT RAS signaling is essential to developing rational therapeutic approaches to treat RAS-mutated cancers. We found that WT RAS enhanced mutant RAS-driven transformation by activating RAS effectors poorly engaged by mutated RAS. Further, inhibition of RAS effectors activated poorly by mutant RAS synergized with and limited resistance to mutant HRAS and KRAS inhibitors. The mutant HRAS inhibitor tipifarnib blocked PI3K signaling and synergized with MEK inhibitors in HRAS-mutated cancer cell lines; covalent KRASG12C inhibitors blocked MEK signaling and synergized with PI3K inhibitors in KRASG12C- mutated cell lines. Synergy between inhibitors of mutant RAS and RAS effectors was dependent on intact RTK/WT RAS signaling and was lost in both RASless or SOSless MEFs. Dual knock-out of WT RAS isoforms in KRAS- and HRAS-mutated cancer cell lines reduced PI3K signaling in KRAS-mutant cell lines and MAPK signaling in HRAS-mutant cell lines and reduced proliferation, confirming our findings in MEF cells. Overall, our data highlight the critical role of WT RAS isoforms in supporting mutant RAS signaling and should be considered when designing combination therapies in RAS-mutated cancers. Citation Format: Nancy E. Sealover, Bridget Finniff, Jacob Hughes, Hyun Lee, Robert L. Kortum. Wild-type RAS signaling is an essential therapeutic target in RAS-mutated cancers [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2024; Part 1 (Regular Abstracts); 2024 Apr 5-10; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2024;84(6_Suppl):Abstract nr 1925.
Abstract 2368: Intratumoral delivery of HER-2-CD3hFc mRNA-LNP inhibits ovarian tumor growth
Authors: Vita M. Golubovskaya, John Sienkiewicz, Jinying Sun, Shiming Zhang, Yanwei Huang, Liang Hu, Hua Zhou, Hizkia Harto, Shirley Xu, Robert Berahovich, Lijun Wu
Journal: Cancer Research (2024)

Abstract Her-2 is a well-known driver of carcinogenesis that is overexpressed in breast, ovarian and other tumors. Immunotherapy with bispecific antibodies is a promising cancer therapy approach. Her-2 (Herceptin) antibody was used to test bispecific antibody against ovarian cancer cells. First, Her-2-CAR-T cells with Her-2 (4D5) ScFv and 4-1BB costimulatory domain were generated and tested using in vitro killing and IFN-gamma assays, and in vivo efficacy SKOV-3 xenograft NSG mouse model. Her-2-CAR-T cells killed SKBR-3 breast and SKOV-3 ovarian cancer cells and stopped SKOV-3 xenograft tumor growth. This Her-2-ScFv was used to generate bispecific Her-2 ScFv-CD3-ScFv human Fc antibody using in vitro transcribed mRNA. The 5’capped Her-2-CD3-hFc mRNA with 5’UTR, 3’UTR and poly A tail was generated by in vitro transcription using DNA template. The mRNA was embedded into lipid nanoparticles (LNP) using microfluidic PreciGenome Flex S NanoGenerator. Western blotting detected &amp;gt;160 kDa bispecific antibody secreted from A1847 and 293 cells transfected with mRNA-LNPs. The bispecific antibody was binding Her-2-positive A1847 ovarian cancer cell lines and T cells by FACS analysis. The Her-2-CD3-hFc bispecific antibody with T cells effectively killed Her-2-positive target cells in a dose-dependent manner in RTCA assay and produced high level of IFN-gamma. Moreover, intratumoral delivery of Her-2-CD3-hFc mRNA-LNP with intravenous injection of T cells completely blocked A1847 xenograft tumor growth. In conclusion. intratumoral delivery of Her-2-CD3 mRNA-LNP blocks ovarian xenograft tumor growth and provides a solid basis for future pre-clinical and clinical studies. Citation Format: Vita M. Golubovskaya, John Sienkiewicz, Jinying Sun, Shiming Zhang, Yanwei Huang, Liang Hu, Hua Zhou, Hizkia Harto, Shirley Xu, Robert Berahovich, Lijun Wu. Intratumoral delivery of HER-2-CD3hFc mRNA-LNP inhibits ovarian tumor growth [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 2368.
Abstract 5109: Novel acylfulvene LP184 shows single agent and combinatorial efficacy with PARP inhibitors in atypical teratoid rhabdoid tumors
Authors: Abigail Tindall, Hyuk J. Kwon, Istabraq Musa, Tyler Findlay, Su Chan Lee, Charles Eberhart, Eric Raabe
Journal: Cancer Research (2024)

Abstract Atypical Teratoid Rhabdoid Tumor (AT/RT) is an aggressive form of pediatric brain cancer that spreads quickly and leads to survival rates of less than 40%. Current treatments are insufficient at reducing this poor prognosis. A hallmark feature of ATRT is a SMARCB1 mutation. Illudins are a class of cytotoxic compounds with activity against pediatric solid tumors, including AT/RT. LP-184 is a novel synthetic illudin with improved brain penetration and activity against adult high-grade glioma orthotopic xenografts. We tested LP-184 against a battery of AT/RT cell lines including BT37, CHLA06, CHLA05, and CHLA266. LP-184 decreases viability at nanomolar concentrations with IC50s ranging from 9.91nM - 23.92nM. LP-184 decreased proliferation as measured by immunofluorescence and confirmed by western blots (BT37 p=0.0001, CHLA06 =0.0001, CHLA05=0.033). LP-184 also induced apoptosis as demonstrated in immunofluorescent images and western blots (BT37 p=0.0001, CHLA06 =0.0001, CHLA05=0.0015, CHLA266 p=0.002). Furthermore, LP-184 shows strong synergy with the PARP inhibitor Rucaparib, suggesting that inhibition of DNA repair potentiates LP-184 activity. LP-184 reduces viable cell count by as much as 50% and combination with Rucaparib further decreases viability by 75% (BT37=0.0001, CHLA06=0.0002, CHLA05=0.0077). The dual therapy also demonstrates cytostatic and cytotoxic effects as confirmed by immunofluorescence and western blots (BT37 BrdU = 0.0004, BT37 CC3 = 0.0002, CHLA06 BrdU = 0.0033, CHLA06 CC3 = 0.0117 compared to control). Further studies in vivo are being conducted to test single agent and combination therapies of LP-184 and Rucaparib in ATRT, as our data indicates that the two could provide a compelling treatment option for patients with this poor prognosis brain tumor. Citation Format: Abigail Tindall, Hyuk J. Kwon, Istabraq Musa, Tyler Findlay, Su Chan Lee, Charles Eberhart, Eric Raabe. Novel acylfulvene LP184 shows single agent and combinatorial efficacy with PARP inhibitors in atypical teratoid rhabdoid tumors [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 5109.