bluuebunny/crossref_metadata_2025_split huggingface dataset:
Abstract 2558: Loss of TC-PTP in keratinocytes leads to increased UVB-induced autophagy
Authors: Asare Obed , Zavala Klarissa , Shim Lindsey , Hafeez Bilal , Kim Dae 
Published: April 2023

Abstract
Amongst the deteriorating effects of the sun’s ultraviolet B (UVB) radiation on cellular homeostasis is the formation of photoproducts in the DNA, such as cyclobutene pyrimidine dimers. Photoproducts cause conformational alterations in the DNA structure which may evade cellular repair machinery and persist through DNA replication processes, leading to potential gene mutations and ultimately leading to skin cancer. Photoproducts are typically repaired by nucleotide excision repair. Notwithstanding, when the repair mechanism fails, apoptosis ensues to prevent the accumulation of mutations and to restore cellular homeostasis. T-cell protein tyrosine phosphatase (TC-PTP) is one of the members of PTP family that has been shown to increase UVB-induced apoptosis in keratinocytes via the downregulation of Flk-1/JNK signaling. This implies that loss of TC-PTP causes resistance to apoptosis in UVB-damaged keratinocytes. In the present work, we report that this resistance is accompanied by an increase in UVB-induced autophagy in the absence of TC-PTP. We generated human TC-PTP-deficient (TC-PTP/KO) HaCaT keratinocytes using CRISPR/Cas9 system. The expression of microtubule-associated protein light chain 3 (LC3) was significantly increased in TC-PTP/KO HaCaT keratinocytes compared to engineered controls (TC-PTP/Mock) following treatment with UVB irradiation. Increased expression of LC3 in TC-PTP/KO keratinocytes was accompanied by a significant decrease in the expression of the p62. We also observed that treatment of TC-PTP KO keratinocytes with chloroquine (CQ), a late-phase autophagy inhibitor, caused a remarkable increase in LC3 expression following UVB exposure in comparison with untreated TC-PTP/KO keratinocytes, implying that loss of TC-PTP increases autophagy in response to UVB. With this regard, TC-PTP deficiency in human keratinocytes significantly increased cell proliferation following UVB exposure compared to similarly treated control keratinocytes. Pretreatment of CQ before UVB irradiation in TC-PTP KO keratinocytes significantly reduced cell viability compared to untreated TC-PTP KO keratinocytes. These results suggest that TC-PTP protects keratinocytes from UVB-induced damage by negatively regulating autophagy signaling.
Citation Format: Obed Asare, Klarissa Zavala, Lindsey Shim, Bilal Hafeez, Dae Kim. Loss of TC-PTP in keratinocytes leads to increased UVB-induced autophagy [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 2558.
Abstract 2605: LOXL3 knock out affects pathways which involve cytoskeleton regulation, proliferation and apoptosis in glioblastoma cells
Authors: Laurentino Talita S. , Soares Roseli S. , Lerario Antonio M. , Marie Suely K. , Oba-Shinjo Sueli Mieko 
Published: April 2023

Abstract
Lysyl oxidase-like 3 (LOXL3), belonging to the lysyl oxidase family of copper-dependent amino oxidase, is responsible for the crosslinking of collagen and elastin. In tumors, the presence of LOXL3 has been associated with genomic stability, cell proliferation, and metastasis. In silico analysis has shown that glioblastoma was among the tumors with the highest LOXL3 expression levels. Moreover, LOXL3 expression increased with malignancy grade amongst diffusely infiltrative astrocytomas (from grade 2 to 4). In the present work, CRISPR-Cas9 gene editing was used to knock out LOXL3 in the human glioma U87MG cell line to evaluate the role of LOXL3 in glioblastoma. Analyses of protein expression level and genomic mutation mediated by CRISPR-Cas9 by two different sgRNA (denominated sgRNA1 and 2) were performed and the knock out efficiency and genome editing were confirmed. Transcriptome analysis of these cells revealed a downregulation of genes that coded for proteins involved in microtubule organization, cell division, and vesicle trafficking. On the other hand, upregulated genes when LOXL3 was knocked out in U87MG cells were involved in actin cytoskeleton, negative regulation of gene transcription, including chromatin remodeling, and cell-cell adhesion. Functional in vitro assays showed that LOXL3 knocked out cells presented decreased cell proliferation due to longer cell cycles with increased G1 and G2/M checkpoints compared to control cells. Additionally, LOXL3 knocked out U87MG cells presented a higher percentage of cells in the early phase of apoptosis with an increment after temozolomide treatment. Our results suggest an important role of LOXL3 in glioblastoma possibly mediated by cytoskeleton regulation, affecting cell proliferation, cell cycle, and cell death. These results reinforced the importance of LOXL3 in tumors, especially in glioblastoma, highlighting it as a potential therapeutic target.
Citation Format: Talita S. Laurentino, Roseli S. Soares, Antonio M. Lerario, Suely K. Marie, Sueli Mieko Oba-Shinjo. LOXL3 knock out affects pathways which involve cytoskeleton regulation, proliferation and apoptosis in glioblastoma 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 2605.
Abstract 2607: Homogenous TP53mut-associated tumor biology across mutation and cancer types revealed by comprehensive mRNA expression analysis
Authors: Budczies Jan , Romanovsky Eva , Kluck Klaus , Ourailidis Iordanis , Menzel Michael , Beck Susanne , Ball Markus , Kazdal Daniel , Christopoulos Petros , Schirmacher Peter , Stiewe Thorsten , Stenzinger Albrecht 
Published: April 2023

Abstract
TP53 mutations are the most common single gene alteration in human cancer with diagnostic and prognostic implications in some cancer types. While no TP53-targeting therapeutics have been approved in the USA or Europe yet, drugs tailored to specific TP53 mutations, restoring the functionality of mutated TP53 (TP53mut), and protecting TP53 from negative regulation are being explored in preclinical studies and clinical trials. We performed a comprehensive mRNA expression analysis in 24 cancer types of the TCGA to extract (i) a consensus expression signature shared across TP53 mutation types and cancer types, (ii) differential gene expression patterns between different TP53 mutation types such as LOF, GOF, as well as dominant-negative mutations, and (iii) cancer types specific gene expression patterns. Mutational hotspots showed a similar pattern across cancer types, but at the same time prevalence was significantly different between cancer types for about half of the most prevalent hotspots. Both can be explained in part by the mutational processes operational for example the clock-like process behind SBS1 that is operational across cancer types and aflatoxin exposure assocated with SBS24 that ist operational in liver hepatocellular carcinoma. Virtually no genes were differential between tumors harboring different types of TP53 mutations in none of the cancer types, while hundreds of genes were over- and underexpressed in TP53mut compared to TP53wt tumors. A consensus gene list of 178 common over- and 32 common underexpressed genes was shared between at least two-thirds of the 24 cancer types. Analysis of the immune tumor microenvironment revealed exclusively decreased immune cell populations in the TP53mut tumors in six, a mixed pattern in four, exclusively increased immune cell populations in two, and no significant alterations in 20 cancer subtypes. The analysis of a large cohort of human tumors complements results from experimental studies and support the development of novel strategies for therapeutic targeting of TP53mut.
Citation Format: Jan Budczies, Eva Romanovsky, Klaus Kluck, Iordanis Ourailidis, Michael Menzel, Susanne Beck, Markus Ball, Daniel Kazdal, Petros Christopoulos, Peter Schirmacher, Thorsten Stiewe, Albrecht Stenzinger. Homogenous TP53mut-associated tumor biology across mutation and cancer types revealed by comprehensive mRNA expression analysis [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 2607.
Abstract 2608: Defining the function of tumor suppressor p53 arginine 181 residue in tumorigenesis
Authors: Moses Renyta , Kelly Gregory , Indeglia Alexandra , Miller Sven , Karanicolas John , Murphy Maureen , Maxwell Kara 
Published: April 2023

Abstract
The gene Tp53 encodes for the protein p53 which has many tumor suppressive functions. Li-Fraumeni syndrome is characterized by germline Tp53 mutations that significantly increase the risk of cancer. The p53 amino acid residue arginine 181 is one such mutation site that promotes tumorigenesis by impacting a phenomenon known as p53 cooperativity. This is the ability of one p53 molecule to interact with another in order to efficiently bind specific sites on DNA to enact specific p53 functional pathways. Two p53 molecules interact with each other through the formation of a salt bridge between H1 helices found in respective DNA binding domains. The two key amino acid residues in this salt bridge are positively charged arginine (R) 181 and negatively charged glutamic acid (E) 180 which form electrostatic interactions with the residues on the opposite p53 H1 helix. Cooperativity mutants such as R181H and R181C show a hypomorphic behavior wherein much of p53’s canonical functions remain intact. Colony suppression data show that H1299 cells with p53 R181H and R181C yield a similar number of colonies as p53 wild type, which is significantly fewer than the yield of p53 hotspot mutation R175H and the absence of p53. Despite retaining some wild-type behavior, R181 mutants are found in LFS patients with cancer. Discovering the importance of the R181 residue for p53’s role in defending against tumorigenesis can lead to unique treatment opportunities for LFS patients with cooperativity mutants.
Citation Format: Renyta Moses, Gregory Kelly, Alexandra Indeglia, Sven Miller, John Karanicolas, Maureen Murphy, Kara Maxwell. Defining the function of tumor suppressor p53 arginine 181 residue in tumorigenesis [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 2608.
Abstract 2619: Characterization of RAS assembly architectures in living cells
Authors: Zhou Zhiwei , Li Xing-Xiao , Meyer Cynthia , Zhang Yu-An , Angira Deekshi , Westover Kenneth 
Published: April 2023

Abstract
Assembly of RAS proteins and RAS effectors into “nanoclusters” at the cell membrane regulates RAS signaling output. The structure and composition of membrane-bound RAS complexes is dynamic, but defined states appear to exist. We previously found biological and biophysical evidence for two different models of membrane-bound RAS assemblies where RAS proteins form symmetric or asymmetric RAS-RAS interactions. It is unknown what functions these two architectures might serve and what determinants these nanoclusters might be subject to. To further characterize which architectural states occur in various biological scenarios, we developed a plate-based assay to detect RAS-RAS interactions in cells. The system controls for RAS expression and can be used to discriminate symmetric and asymmetric RAS-RAS interactions. It can also be used to assess the relative contributions of the G domain (1-166) and hypervariable region (HVR) in mediating RAS-RAS interactions. We found that RAS isoforms can homo- and hetero-dimerize and observed that the G domain contributes to RAS-RAS interactions, but that the HVR does not. We also found that the primary determinant of HVR interaction with the membrane is electrostatic charge. Regarding cancer-causing mutations, we found that in the KRAS G12D context, RAS-RAS interactions appear to favor asymmetric RAS-RAS interactions while KRAS Q61H favors symmetric RAS-RAS interactions.
Citation Format: Zhiwei Zhou, Xing-Xiao Li, Cynthia Meyer, Yu-An Zhang, Deekshi Angira, Kenneth Westover. Characterization of RAS assembly architectures in living 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 2619.
Abstract 2646: Screening the membrane proteome to determine antibody specificity and de-risk CAR-T cell development
Authors: Rucker Joseph , Fong Rachel H. , Sullivan Tabb , Navia Carmen , Screnci Brad , Wilf Rona , Doranz Benjamin J. 
Published: April 2023

Abstract
Rigorous specificity analysis is critical for the development of antibody-based therapies, as even minimal off-target binding can lead to toxicity and clinical failures. Long believed to be exquisitely specific, recent preclinical data and our own work indicate that monoclonal antibodies (MAbs) frequently (~25%) display cross-reactivity. In many cases, off-target interactions occur with unrelated proteins that cannot be predicted by protein sequence homology. Cross-reactivity can lead to serious or even life-threatening consequences especially when MAbs are configured as bispecifics, antibody-drug conjugates (ADC) or CAR-T cell therapies, and IND applications for biotherapeutics require cross-reactivity assessment to prevent adverse events. Tissue cross-reactivity (TCR) studies have traditionally been used to screen for off-target binding, however, with poor predictive value for in vivo safety and toxicity. We developed the Membrane Proteome Array (MPA) platform to de-risk MAb-based therapeutics by testing for specificity across 6,000 human membrane proteins expressed in live cells. In contrast to TCR studies, proteins in the MPA exist in their native conformations and are not altered by fixation. The MPA assesses binding interactions by high-throughput flow cytometry allowing for high sensitivity detection and rapid analysis. We will discuss the importance of early-stage specificity testing to expose possible toxicities and present case studies for antibody and cell therapy profiling. MPA data have been used in numerous successful regulatory applications and may be used to replace or complement other cross-reactivity studies. In particular, MPA profiling has been used in IND applications for CAR-T and cell-therapies where conventional immunohistochemical binding assays did not suffice.
Citation Format: Joseph Rucker, Rachel H. Fong, Tabb Sullivan, Carmen Navia, Brad Screnci, Rona Wilf, Benjamin J. Doranz. Screening the membrane proteome to determine antibody specificity and de-risk CAR-T cell development [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 2646.
Abstract 2686: The combination of Cucurbitacin B and Evodiamine inhibits cell migration and propagation, and induces apoptosis in in vitro glioma cells
Authors: Mehta Vrutti V. , Benzeroual Kenza E. 
Published: April 2023

Abstract
Glioblastoma is a lethal brain tumor. Despite aggressive treatment, the median survival time is 6 to 12 months. These tumors acquire resistance against many therapeutic regimens. There is a need to screen for potential new drugs and drug combinations. Cucurbitacin B (CuB), a compound obtained from plants, and evodiamine (Evo), a supplement, have shown promising results in various neoplasms but have not been well studied in glioblastoma and especially, as a combination treatment. This study investigated the combined effects of CuB and Evo on U-87MG human glioma cell line. Results demonstrated that CuB and Evo individually decreased the U-87MG cell viability in time and dose dependent manners. The effect was significantly pronounced with the combined treatment. The combination treatment of CuB and Evo induced cellular apoptosis most likely by interfering with the PI3K/AKT and JNK/ERK pathways. In addition, the flow cytometry analysis showed that the combination treatment arrested the cell cycle at the G2M phase and inhibited cell propagation. Moreover, CuB and Evo combined effect successfully inhibited cellular migration. The cytotoxic effect of combination treatment of CuB and Evo was three times lower in neuronal PC-12 cells compared to U-87MG cells. Overall, a combination of cucurbitacin B and evodiamine proved to be a superior potential anti-neoplastic treatment in U-87MG cells as compared to individual treatment, and may be considered as a potential adjuvant therapy for GBM.
Citation Format: Vrutti V. Mehta, Kenza E. Benzeroual. The combination of Cucurbitacin B and Evodiamine inhibits cell migration and propagation, and induces apoptosis in in vitro glioma 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 2686.
Abstract 2694: Blockade of the cyclooxygenase-2 prevents chemotherapy-induced cognitive impairments
Authors: Rashid Mohammad Abdur , Tang Jason J. , Yoo Ki-Hyun , Corujo-Ramirez Ana , Kim Sang Hoon , Oliveros Alfredo , Cole Peter , Hawse John R. , Jang Mi-Hyeon 
Published: April 2023

Abstract
Chemotherapy-induced cognitive impairment (CICI, also termed “chemobrain”) is a major neurotoxic side effect exhibited by a wide range of chemotherapeutic agents. These side effects affect approximately 14 million cancer survivors. More importantly, CICI persists well after cessation of therapy, severely interfering with quality of life. Unfortunately, there is no treatment and novel therapies are urgently needed. Utilizing cisplatin, a platinum-based chemotherapy, to model chemobrain mice in vivo, we have established that cisplatin accelerates the brain aging process thus leading to long-term memory impairment in mice similar to what is clinically reported. We further show that cisplatin causes oxidative DNA damage, mitochondrial defects, impaired neurogenesis, synaptic defects, and increased gliosis in the adult hippocampus, a brain region critical for learning and memory. Mechanistically, our RNA-sequencing and qRT-PCR analysis revealed that cisplatin dramatically increases COX-2 (Ptgs2) expression and its major product, prostaglandin E2 (PGE2) in adult mouse brain and human cortical neurons derived from induced pluripotent stem cells (iPSCs). Similar as cisplatin treatment, the levels of COX-2 expression began to increase at 0.1 µM methotrexate treatment, which significantly increased at 1 µM in human cortical neurons, indicating that COX-2 induction is a common pathogenic mechanism mediating cognitive impairment associated with cisplatin and methotrexate in spite of the fact that these compounds have different mechanisms of action for cancer eradication. Most importantly, NS-398, a selective COX-2 inhibitor, effectively prevents cognitive deficits in mice and reduction in cell viability of human neurons associated with these chemotherapies, without promoting tumor growth or interfering with cisplatin’s anti-tumor activity. Taken together, our findings strongly suggest that COX-2 induction is a main cause of CICI, making COX-2 inhibition a therapeutic target for CICI. Given that COX-2 inhibitors are currently being tested in cancer prevention as well as age-related memory loss in clinical trials, inhibiting COX-2 may have far-reaching therapeutic effects on CICI and cancer treatment, which can be applied quickly, safely, and effectively to clinical trials, in the effort to improve cancer survivor quality of life.
Citation Format: Mohammad Abdur Rashid, Jason J. Tang, Ki-Hyun Yoo, Ana Corujo-Ramirez, Sang Hoon Kim, Alfredo Oliveros, Peter Cole, John R. Hawse, Mi-Hyeon Jang. Blockade of the cyclooxygenase-2 prevents chemotherapy-induced cognitive impairments [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 2694.
Abstract 2705: Low-dose crystalline cytokine macrophage mediated Trojan horse immunotherapy of solid cancers
Authors: Jones Michael H. , Calla Nirk E. Quispe , Talbot-Cooper Callum , Zeng Hong , Best Jonathan , Smith Robert 
Published: April 2023

Abstract
Protein therapeutics are often compromised by sub-optimal biodistribution contributing to poor efficacy and adverse events. Drug delivery mechanisms better able to target the disease site and provide localized, sustained drug release have the potential to transform therapeutic standards. PODS crystals (PCs) are natural-mimetic, micron-scale protein co-crystals engineered to incorporate a protein cargo that is sustainably released under the action of resident proteases. PCs are efficiently taken up by phagocytic cells and their cargo protein is subsequently released in a bioactive form. Since blood-circulating phagocytic cells, including monocytes, are actively recruited into the tumor microenvironment, we postulated that monocyte/macrophage-mediated PC delivery could be used as a molecular “Trojan horse” to efficiently deliver therapeutic proteins to target cells. This could improve the pharmacodynamics and pharmacokinetics of protein drug delivery to treat systemic and disseminated diseases. Interleukin-2 (IL-2) is notoriously toxic at the high doses required for therapeutic efficacy. Here, we demonstrate the therapeutic efficacy and tolerability of low doses of PC-IL-2 administered intravenously in a mouse model of melanoma. We further demonstrate the therapeutic benefit of PCs delivering low doses of IL-2, interleukin-15 (IL-15) and interferon-gamma (IFN-γ) in a mouse model of renal cell carcinoma. In all the treated mice, cachexia, a side effect of high-dose cytokine therapy, was notably absent. This study provides proof-of-concept for the utility of intravenously administered PCs to provide a generalised and widely applicable mechanism to effectively deliver protein drugs for the therapy of cancer and other diseases.
Citation Format: Michael H. Jones, Nirk E. Quispe Calla, Callum Talbot-Cooper, Hong Zeng, Jonathan Best, Robert Smith. Low-dose crystalline cytokine macrophage mediated Trojan horse immunotherapy of solid cancers [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 2705.