bluuebunny/crossref_metadata_2025_split huggingface dataset:
Evaluation of a commercial proAKAP4 kit for the assessment of fresh and frozen–thawed feline spermatozoa
Authors: Prochowska Sylwia , Eberhardt Maria , Niżański Wojciech 
Published: March 2024

AbstractProAKAP4 is gaining increasing attention as a potential marker of semen quality in many species, but while there is a commercial kit for assessing proAKAP4 in the domestic cat, there are no publications about its use. The aim of this study was to evaluate the commercial proAKAP4 kit – Cat 4MID® Kit (SPQI – 4BioDx, Lille, France) for the assessment of feline semen. Semen was collected from 54 male cats by urethral catheterization. After a basic semen evaluation (subjective motility, CASA, viability, morphology), proAKAP4 levels in each sample were assessed using the Cat 4MID® Kit according to the manufacturer's protocol or with some modifications related to incubation time, sample storage conditions and number of spermatozoa used. Finally, the Spearman correlation of proAKAP4 concentration and sperm motility parameters was calculated. The most reliable results (acceptable intraassay coefficient of variation) were obtained with an optimized protocol of overnight incubation and isolation of proAKAP4 protein from 1 × 106 spermatozoa stored at −80°C. For fresh semen, there were no significant correlations between proAKAP4 concentration and sperm motility parameters, despite a strong correlation between motility parameters and sperm viability and morphology. A predominant effect of other sperm parameters and highly variable performance of lysis buffer question the usefulness of Cat 4MID® Kit for the assessment of feline semen. For frozen–thawed semen, there was a moderate, negative correlation between proAKAP4 concentration and two CASA parameters, VAP and VSL. As there were no correlations between proAKAP4 concentration in fresh semen and motility parameters in cryopreserved samples, proAKAP4 cannot be used as freezability marker in cats. More studies are needed to establish potential correlation with long‐lasting motility.
Impact of surgical lung biopsy on lung function and survival in patients with idiopathic pulmonary fibrosis in a multi‐centre registry cohort
Authors: Marcoux Veronica , Lok Stacey D. , Mondal Prosanta , Assayag Deborah , Fisher Jolene H. , Shapera Shane , Morisset Julie , Manganas Hélène , Fell Charlene D. , Hambly Nathan , Cox P. Gerard , Kolb Martin , Gershon Andrea S. , To Teresa , Sadatsafavi Mohsen , Khalil Nasreen , Wong Alyson W. , Wilcox Pearce G. , Ryerson Christopher J. , Vu Thao , Johannson Kerri A. 
Published: March 2024

AbstractBackground and ObjectiveEstablishing an accurate and timely diagnosis of idiopathic pulmonary fibrosis (IPF) is essential for appropriate management and prognostication. In some cases, surgical lung biopsy (SLB) is performed but carries non‐negligible risk. The objective of this retrospective study was to determine if SLB is associated with accelerated lung function decline in patients with IPF using the Canadian Registry for Pulmonary Fibrosis.MethodsLinear mixed models and Cox proportional hazards regression models were used to compare decline in forced vital capacity (FVC)%, diffusion capacity of the lung (DLCO%) and risk of death or lung transplantation between SLB and non‐SLB patients. Adjustments were made for baseline age, sex, smoking history, antifibrotic use, and lung function. A similar analysis compared lung function changes 12 months pre‐ and post‐SLB.ResultsA total of 81 SLB patients and 468 non‐SLB patients were included. In the SLB group, the post‐biopsy annual FVC% decline was 2.0% (±0.8) in unadjusted, and 2.1% (±0.8) in adjusted models. There was no difference in FVC% decline, DLCO% decline, or time to death or lung transplantation between the two groups, in adjusted or unadjusted models (all p‐values >0.07). In the pre‐post SLB group, no differences were identified in FVC% decline in unadjusted or adjusted models (p = 0.07 for both).ConclusionNo association between SLB and lung function decline or risk of death or lung transplantation was identified in this multi‐centre study of patients with IPF.image
Adipose Stromal Cell–Derived Cancer-Associated Fibroblasts Suppress FGFR Inhibitor Efficacy
Authors: Kolonin Mikhail G. , Anastassiou Dimitris 
Published: March 2024

Abstract
Cancer aggressiveness has been linked with obesity, and studies have shown that adipose tissue can enhance cancer progression. In this issue of Cancer Research, Hosni and colleagues discover a paracrine mechanism mediated by adipocyte precursor cells through which urothelial carcinomas become resistant to erdafitinib, a recently approved therapy inhibiting fibroblast growth factor receptors (FGFR). They identified neuregulin 1 (NRG1) secreted by adipocyte precursor cells as an activator of HER3 signaling that enables resistance. The NRG1-mediated FGFR inhibitor resistance was amenable to intervention with pertuzumab, an antibody blocking the NRG1/HER3 axis. To investigate the nature of the resistance-associated NRG1-expressing cells in human patients, the authors analyzed published single-cell RNA sequencing data and observed that such cells appear in a cluster assigned as inflammatory cancer-associated fibroblasts (iCAF). Notably, the gene signature corresponding to these CAFs is highly similar to that shared by adipose stromal cells (ASC) in fat tissue and fibro-adipogenic progenitors (FAP) in skeletal muscle of cancer-free individuals. Because fibroblasts with the ASC/FAP signature are enriched in various carcinomas, it is possible that the paracrine signaling conferred by NRG1 is a pan-cancer mechanism of FGFR inhibitor resistance and tumor aggressiveness.
See related article by Hosni et al., p. 725
A Pre-Leukemic DNA Methylation Signature in Healthy Individuals at Higher Risk for Developing Myeloid Malignancy
Authors: Lao Zhentang , Ding Ling-Wen , Sun Qiao-Yang , Jia Li , Yan Benedict , Ng Alvin Yu-Jin , Capinpin Sharah Mae , Wang Renwei , Ying Li , Chng Wee Joo , Koeffler H. Phillip , Koh Woon-Puay , Yuan Jian-Min , Yang Henry , Goh Yeow Tee , Grigoropoulos Nicholas 
Published: March 2024

AbstractPurpose:DNA methylation alterations are widespread in acute myelogenous leukemia (AML) and myelodysplastic syndrome (MDS), some of which appear to have evolved independently of somatic mutations in epigenetic regulators. Although the presence of somatic mutations in peripheral blood can predict the risk of development of AML and MDS, its accuracy remains unsatisfactory.Experimental Design:We performed global DNA methylation profiling in a case control study nested within the Singapore Chinese Health Study to evaluate whether DNA methylation alterations were associated with AML/MDS development. Targeted deep sequencing and methylated DNA immunoprecipitation sequencing (MeDIP-seq) were performed on peripheral blood collected a median of 9.9 years before diagnosis of AML or MDS, together with age-matched still-healthy individuals as controls.Results:Sixty-six individuals who developed AML or MDS displayed significant DNA methylation changes in the peripheral blood compared with 167 age- and gender-matched controls who did not develop AML/MDS during the follow-up period. Alterations in methylation in the differentially methylation regions were associated with increased odds of developing AML/MDS.Conclusions:The epigenetic changes may be acquired independently and before somatic mutations that are relevant for AML/MDS development. The association between methylation changes and the risk of pre-AML/MDS in these individuals was considerably stronger than somatic mutations, suggesting that methylation changes could be used as biomarkers for pre-AML/MDS screening.
The Irreversible FGFR Inhibitor KIN-3248 Overcomes FGFR2 Kinase Domain Mutations
Authors: Balasooriya Eranga R. , Wu Qibiao , Ellis Haley , Zhen Yuanli , Norden Bryanna L. , Corcoran Ryan B. , Mohan Adithi , Martin Eric , Franovic Aleksandra , Tyhonas John , Lardy Matthew , Grandinetti Kathryn B. , Pelham Robert , Soroceanu Liliana , Silveira Vanessa S. , Bardeesy Nabeel 
Published: March 2024

Abstract

Purpose:
FGFR2 and FGFR3 show oncogenic activation in many cancer types, often through chromosomal fusion or extracellular domain mutation. FGFR2 and FGFR3 alterations are most prevalent in intrahepatic cholangiocarcinoma (ICC) and bladder cancers, respectively, and multiple selective reversible and covalent pan-FGFR tyrosine kinase inhibitors (TKI) have been approved in these contexts. However, resistance, often due to acquired secondary mutations in the FGFR2/3 kinase domain, limits efficacy. Resistance is typically polyclonal, involving a spectrum of different mutations that most frequently affect the molecular brake and gatekeeper residues (N550 and V565 in FGFR2).

Experimental Design:
Here, we characterize the activity of the next-generation covalent FGFR inhibitor, KIN-3248, in preclinical models of FGFR2 fusion+ ICC harboring a series of secondary kinase domain mutations, in vitro and in vivo. We also test select FGFR3 alleles in bladder cancer models.

Results:
KIN-3248 exhibits potent selectivity for FGFR1–3 and retains activity against various FGFR2 kinase domain mutations, in addition to being effective against FGFR3 V555M and N540K mutations. Notably, KIN-3248 activity extends to the FGFR2 V565F gatekeeper mutation, which causes profound resistance to currently approved FGFR inhibitors. Combination treatment with EGFR or MEK inhibitors potentiates KIN-3248 efficacy in vivo, including in models harboring FGFR2 kinase domain mutations.

Conclusions:
Thus, KIN-3248 is a novel FGFR1–4 inhibitor whose distinct activity profile against FGFR kinase domain mutations highlights its potential for the treatment of ICC and other FGFR-driven cancers.
Abstract B039: Identification of sex-specific molecular features in glioma transcriptome
Authors: Huang Yingbo , Shan Yuting , Zhang Weijie , Printzis Christina , Pesce Lorenzo , Stranger Barbara Elaine , Huang R. Stephanie 
Published: March 2024

Abstract
Glioma is a lethal disease with limited treatment options available. Sex differences in glioma are well-documented in clinical settings, with males having twice the risk of developing the disease and increased mortality compared to female counterparts. To investigate the molecular basis of sex differences in glioma, we analyzed transcriptomic sex differences in a collection of three independent glioma datasets and validated them in additional independent studies. The impact of differentially expressed genes were assessed through Gene Set Enrichment Analysis (GSEA) on approximately 5000 gene sets to interpret their biological relevance. Given the well-recognized heterogeneity of glioma, we also integrated three single-cell RNA (scRNA) sequencing datasets to decipher the sex differences in cellular composition and cell-type specific differential gene expression between male and female tumors. The effects of sex on gene expression are modest and primarily driven by the X chromosome (14 out of 16 differentially expressed genes were on the X chromosome). These genes are associated with escape from X-chromosome inactivation (e.g., KDM5C, KDM6A) and are also sex-differentially expressed (DE) in the healthy human brain according to Genotype-Tissue Expression (GTEx) data. GSEA results suggest increased activity in cell proliferation and neuronal signaling in male GBM samples, whereas females show enrichment in glycoprotein/lipid metabolism. By deconvoluting the cell-type composition, we identified a higher percentage of pericytes in the male glioma tumor microenvironment compared to those in females, which is consistent with recent spatial-omic discoveries in glioma. By integrating scRNA sequencing data, we found that both differentially expressed genes and pathways show cell-type-specific enrichment. For example, the female-biased gene GBGT1 is exclusively expressed in the myeloid lineage, while male-biased neuronal pathways are enriched in cancer stem cells. Integrating with the inference of cell trajectories, we observed that male tumors harbor more unconnected, invasive malignant cells than females, which might explain the poor prognosis in male glioma patients. In summary, our results provide a comprehensive multi-level characterization of sex differences in glioma tumors, and these findings will provide novel insights into glioma disease progression and development of new therapy.
Citation Format: Yingbo Huang, Yuting Shan, Weijie Zhang, Christina Printzis, Lorenzo Pesce, Barbara Elaine Stranger, R. Stephanie Huang. Identification of sex-specific molecular features in glioma transcriptome [abstract]. In: Proceedings of the AACR Special Conference on Brain Cancer; 2023 Oct 19-22; Minneapolis, Minnesota. Philadelphia (PA): AACR; Cancer Res 2024;84(5 Suppl_1):Abstract nr B039.
Abstract PR-008: Modeling epigenetic lesions that cause gliomas
Authors: Rahme Gilbert J. , Javed Nauman M. , Puorro Kaitlyn L. , Xin Shouhui , Hovestadt Volker , Johnstone Sarah E. , Bernstein Bradley E. 
Published: March 2024

Abstract
Epigenetic lesions that disrupt gene regulatory elements and expression are increasingly recognized as potential drivers of human cancers. However, we currently lack the in vitro and in vivo models required to functionally validate such lesions and their tumorigenic impact. Here we model aberrations that arise in Isocitrate Dehydrogenase (IDH) mutant lower-grade gliomas, which exhibit profound DNA hypermethylation as a direct consequence of mutant IDH. DNA hypermethylation may promote gliomagenesis by silencing tumor suppressor genes or, alternatively, by activating proto-oncogenes through disruption of CCCTC-binding factor (CTCF) insulators. CTCF insulator sites define the three-dimensional shape of the genome by dictating the boundaries of topologically associated domains (TADs). Enhancers and promoters can interact when located in the same TAD but are restricted from interacting across different TADs. In IDH mutant gliomas, CpG sites around CTCF binding sites are frequently methylated, effectively compromising CTCF binding and thus TAD organization, allowing for cross-TAD interactions and aberrant activation of genes. We discovered a CTCF insulator downstream of the PDGFRA proto-oncogene that is recurrently disrupted in IDH mutant gliomas. We demonstrate that disruption of the syntenic insulator in mouse oligodendrocyte progenitor cells (OPCs) allows an OPC-specific enhancer to contact and induce Pdgfra, thereby increasing proliferation. In contrast, insulator disruption did not affect Pdgfra expression in neural progenitor cells (NPCs), which lack the enhancer. We also model a second recurrent epigenetic lesion in IDH mutant gliomas, the methylation-dependent silencing of the CDKN2A tumor suppressor. We show that inactivation of Cdkn2a/p19ARF by de novo promoter methylation or mutation drives OPC proliferation and cooperates with Pdgfra insulator loss. Finally, we use lentiviruses to coordinately inactivate the Pdgfra insulator and Cdkn2a in mouse corpus callosum, resulting in low-grade gliomagenesis in vivo with histological features reminiscent of human IDH mutant gliomas. Our study recapitulates recurrent epigenetic lesions in mouse models and demonstrates that the combination of Pdgfra activation and Cdkn2a silencing can transform OPCs in vitro and drive gliomagenesis in vivo.
Citation Format: Gilbert J. Rahme, Nauman M. Javed, Kaitlyn L. Puorro, Shouhui Xin, Volker Hovestadt, Sarah E. Johnstone, Bradley E. Bernstein. Modeling epigenetic lesions that cause gliomas [abstract]. In: Proceedings of the AACR Special Conference on Brain Cancer; 2023 Oct 19-22; Minneapolis, Minnesota. Philadelphia (PA): AACR; Cancer Res 2024;84(5 Suppl_1):Abstract nr PR-008.
Abstract A021: The beta-arrestin1/endothelin axis bolsters a CAF-dependent invadosome activity and ovarian cancer metastatic potential
Authors: Rio Danila Del , Masi Ilenia , Ottavi Flavia , Salvati Erica , Caprara Valentina , Bagnato Anna , Giannitelli Sara , Spadaro Francesca , Rosanò Laura 
Published: March 2024

Abstract
The recruitment of fibroblasts to tumors and their activation to cancer-associated fibroblasts (CAFs) is a strategy used by tumor cells to guide extracellular matrix (ECM) remodeling, invasion, and metastasis, highlighting the need to dissect molecular mechanisms involved in CAF function. Endothelin-1 (ET-1) regulates cancer/stroma crosstalk, supporting serous ovarian cancer (SOC) progression. By binding its receptors ETA and ETB, ET-1 allows the recruitment of β-arrestin1 (β-arr1) and the generation of signaling complexes regulating tumor progression. However, how ET-1 receptors might “educate” ovarian fibroblasts (HOFs) to supply altered ECM needs and bolster metastasis is to be investigated. This study identifies ET-1 as a critical factor in activating CAFs capable of proteolytic ECM remodeling by invadosome and creating heterotypic spheroids with cancer cells with favorable metastatic abilities. An autocrine/paracrine ET-1/ETA/BR/β-arr1 loop promotes HOF proliferation, an increased CAF marker expression, secretion of proinflammatory cytokines, collagen contraction ability, and motility. Moreover, ET-1 allows ECM remodeling by supporting the lytic activity of invadosome and activation of integrin beta1. Of note, ET-1 signaling favors the formation of heterotypic HOF/SOC spheroids with enhanced ability to survive, migrate through mesothelial monolayer mesothelial, and invade, representing metastatic units. Blockade of ETA/BR or β-arr1 silencing prevents CAF activation, invadosome function, mesothelial clearance, and invasive ability of heterotypic spheroids. In vivo, therapeutical inhibition of ETA/BR with bosentan markedly reduced the metastatic potential of HOF/SOC spheroids. Our findings support a model in which ET-1/β-arr1 amplifies tumor/stroma interaction via CAF activation and fuels HG-SOC metastatic properties, which might be hampered by ETA/BR antagonists.
Citation Format: Danila Del Rio, Ilenia Masi, Flavia Ottavi, Erica Salvati, Valentina Caprara, Anna Bagnato, Sara Giannitelli, Francesca Spadaro, Laura Rosanò. The beta-arrestin1/endothelin axis bolsters a CAF-dependent invadosome activity and ovarian cancer metastatic potential [abstract]. In: Proceedings of the AACR Special Conference on Ovarian Cancer; 2023 Oct 5-7; Boston, Massachusetts. Philadelphia (PA): AACR; Cancer Res 2024;84(5 Suppl_2):Abstract nr A021.
Abstract A032: The role of ASIC2 and calcium influx in epithelial ovarian cancer pathogenesis
Authors: Joshi Tanvi , Wilhite Annelise , Chokshi Sagar , Singleton Mary Howard , Scalici Jennifer , Lee Kevin 
Published: March 2024

Abstract
Objective: Inflammation associated with incessant ovulation plays a key role in epithelial ovarian cancer (EOC) pathogenesis. Ion channels, such as acid-sensing ion channel-2 or ASIC2 is known to be upregulated in inflammatory conditions and may play a role in calcium ion dysregulation. Here, we describe a potential relationship between ASIC2 and intracellular calcium levels in EOC development and progression. Methods: EOC cell lines OVCAR-8, ES-2 and SKOV-3 were grown to sub-confluence prior to experimentation. Global extracellular pH was changed with the use of hydrochloric acid (HCl) over various time points at hours (h): 2 h, 4 h, 8 h, 12 h and 24 h. ASIC2 expression was subsequently measured via western blot. Intracellular pH was measured with the use of pH-sensitive pHrodo red dye. Cytosolic pH was determined across all cell lines at baseline and after treatment with a known ASIC2 inhibitor: Diminazene (10 μM) over 30 minutes. Intracellular calcium levels were measured with a Fluo Calcium Indicator at baseline and with similar ASIC2 inhibition. Live cell imaging was utilized on a Nikon A1r confocal microscope. Results: Changing global, extracellular pH to 6.0 and 6.5 across various time points from 2-24 h did not affect ASIC2 protein expression via western blot analysis. We then utilized a pH-sensitive assay to determine whether intracellular pH changes were associated with ASIC2 expression. Interestingly, intracellular pH was noted to increase across most cell lines with ASIC2 inhibition via Diminazene treatment. There was a statistically significant increase in pH in ES2 and OVCAR8 cell lines. Because ASIC2 is known to result in influx of various ions, assessment of intracellular calcium levels was performed first after 30 minutes of Diminazene treatment. There was a corresponding significant increase in intracellular calcium levels in ES2 cells after ASIC2 inhibition. And there was a small trend, albeit not significant, toward a similar increase across SKOV-3 and OVCAR-8 cells. Conclusion: There seems to be a role for ASIC2 in EOC, particularly in the context of inflammation as noted by changes in intracellular pH levels. It is likely that ASIC2 may mediate changes in intracellular calcium signaling and promote tumorigenesis in ovarian cancer.
Citation Format: Tanvi Joshi, Annelise Wilhite, Sagar Chokshi, Mary Howard Singleton, Jennifer Scalici, Kevin Lee. The role of ASIC2 and calcium influx in epithelial ovarian cancer pathogenesis [abstract]. In: Proceedings of the AACR Special Conference on Ovarian Cancer; 2023 Oct 5-7; Boston, Massachusetts. Philadelphia (PA): AACR; Cancer Res 2024;84(5 Suppl_2):Abstract nr A032.