Pub Date : 2025-08-25DOI: 10.1016/j.labinv.2025.104232
Lydia Kirsche , Lars Jansen , Annett Petzold , Petra Reinecke , Peter Behrens , Carol Geppert , Nikolaus Gaßler , Matthias Dürst
Human papillomavirus (HPV) DNA integration into the host genome is a frequent event in cervical carcinogenesis and may drive clonal expansion of the affected cells. Based on viral cellular junction (vcj) sequences, highly specific vcj-PCRs can be designed to detect viral integrants in DNA from cervical cell scrapes or tissue samples. In a recent study, such patient-specific vcj-PCR assays were employed for the detection of recurrent high-grade squamous intraepithelial lesions (HSIL) during postoperative surveillance. Although the specificity of vcj-PCR was 100%, only 50% of the recurrences were detected using this approach. The focus of the current study was to analyze the cause of this limited sensitivity. Using chemical microdissection and subsequent vcj-PCR analysis, we could demonstrate that the majority of lesions have a heterogeneous integrant pattern. Only 2 of 16 cones showed a homogeneous distribution of the respective integrants throughout the entire lesion. The other lesions displayed clonal outgrowths harboring the integrant in a background HPV16/18 DNA-positive HSIL tissue. In 4 cases, the respective integrant was undetectable in the lesion. These findings indicate that vcj-PCR has limited sensitivity for the detection of recurrent disease owing to intralesional heterogeneity. The observed heterogeneous integrant pattern may thus reflect the multifocal nature of most large HSIL. Alternatively, the possibility that HPV integration may be a late event in the carcinogenic process also needs to be considered.
{"title":"Heterogeneous Distribution of Human Papillomavirus (HPV) Integration Sites in Cervical Precancers Compromises the Diagnostic Accuracy of Integrant-Specific PCR","authors":"Lydia Kirsche , Lars Jansen , Annett Petzold , Petra Reinecke , Peter Behrens , Carol Geppert , Nikolaus Gaßler , Matthias Dürst","doi":"10.1016/j.labinv.2025.104232","DOIUrl":"10.1016/j.labinv.2025.104232","url":null,"abstract":"<div><div>Human papillomavirus (HPV) DNA integration into the host genome is a frequent event in cervical carcinogenesis and may drive clonal expansion of the affected cells. Based on viral cellular junction (vcj) sequences, highly specific vcj-PCRs can be designed to detect viral integrants in DNA from cervical cell scrapes or tissue samples. In a recent study, such patient-specific vcj-PCR assays were employed for the detection of recurrent high-grade squamous intraepithelial lesions (HSIL) during postoperative surveillance. Although the specificity of vcj-PCR was 100%, only 50% of the recurrences were detected using this approach. The focus of the current study was to analyze the cause of this limited sensitivity. Using chemical microdissection and subsequent vcj-PCR analysis, we could demonstrate that the majority of lesions have a heterogeneous integrant pattern. Only 2 of 16 cones showed a homogeneous distribution of the respective integrants throughout the entire lesion. The other lesions displayed clonal outgrowths harboring the integrant in a background HPV16/18 DNA-positive HSIL tissue. In 4 cases, the respective integrant was undetectable in the lesion. These findings indicate that vcj-PCR has limited sensitivity for the detection of recurrent disease owing to intralesional heterogeneity. The observed heterogeneous integrant pattern may thus reflect the multifocal nature of most large HSIL. Alternatively, the possibility that HPV integration may be a late event in the carcinogenic process also needs to be considered.</div></div>","PeriodicalId":17930,"journal":{"name":"Laboratory Investigation","volume":"105 11","pages":"Article 104232"},"PeriodicalIF":4.2,"publicationDate":"2025-08-25","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144959191","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2025-08-19DOI: 10.1016/j.labinv.2025.104214
Claudio Luchini , Carlotta Franzina , Federico Caldart , Nicolò De Pretis , Manola Crestani , Massimo Donadelli , Paola Mattiolo , Alessandra Fiore , Federica Danzi , Riccardo De Robertis , Michele Bevere , Roberto Baldan , Laura Tommasi , Nicolò Vianini , Paolo Bernardi , Mirco Galiè , Antonio Pea , Rachele Ciccocioppo , Mirko D’Onofrio , Roberto Salvia , Luca Frulloni
Obesity-related diseases and perturbations of fat metabolism represent some of the most common health challenges. In this complex scenario, recent evidence has suggested the emergence of a condition related to fat accumulation in the pancreas, which is generally referred to as fatty pancreas disease. This study aimed to clarify the different compartments of intrapancreatic fat deposition. The study cohort is represented by 100 patients who underwent pancreatic surgical resection. The pancreatic neck margin was analyzed with hematoxylin and eosin for evaluating tissue composition and with Oil Red O, a fat-specific histochemical staining, highlighting lipid droplets as red signals, for evaluating the presence of intracellular fat. Two cases were also analyzed with electron microscopy as cross-sectional validation. Regarding tissue composition, the most prevalent component was normal pancreatic parenchyma (mean value, 71.8%), followed by fibrosis (17.3%) and interlobular/intralobular fat (10.9%). Regarding intracellular fat deposition, Oil Red O–positive intracytoplasmic lipid droplets were present in most patients. The tissue areas with the highest levels of fat deposition were Langerhans’ islets, with neuroendocrine/insular cells showing more commonly a diffuse pattern of fat accumulation (>75% of cells). Electron microscopy confirmed the presence of intracytoplasmic lipid vacuoles in neuroendocrine/insular cells. Our findings showed the presence of different compartments of intrapancreatic fat deposition, both in terms of tissue composition and intracellular compartmentalization. Understanding the mechanisms of fat deposition in the pancreas is crucial toward improving the general knowledge on fatty pancreas disease, also opening new perspectives for the study of lipid metabolism and the treatment of fat-related diseases.
肥胖相关疾病和脂肪代谢紊乱是一些最常见的健康挑战。在这种复杂的情况下，最近的证据表明出现了一种与胰腺脂肪堆积有关的疾病，通常被称为脂肪性胰腺疾病。本研究旨在阐明胰腺内脂肪沉积的不同区室。该研究队列由100例接受胰腺手术切除的患者代表。胰颈缘用苏木精-伊红染色评估组织组成，用Oil Red O（一种脂肪特异性组织化学染色，突出脂滴作为红色信号）评估细胞内脂肪的存在。两个病例也用电子显微镜分析作为横断面验证。在组织组成方面，最常见的是正常胰腺实质（平均值：71.8%），其次是纤维化（17.3%）和小叶间/小叶内脂肪（10.9%）。关于细胞内脂肪沉积，大多数患者存在油红o阳性的胞浆内脂滴。脂肪沉积水平最高的组织区域是朗格汉斯胰岛，神经内分泌/岛细胞更常见地表现为弥漫性脂肪堆积（约占细胞的75%）。电镜检查证实在神经内分泌/岛细胞中存在胞浆内脂泡。我们的研究结果表明，在组织组成和细胞内区隔化方面，胰腺内脂肪沉积存在不同的区隔。了解胰腺脂肪沉积的机制对于提高对脂肪性胰腺疾病的认识至关重要，也为脂质代谢的研究和脂肪相关疾病的治疗开辟了新的视角。
{"title":"Fatty Pancreas Disease: An Integrated Study on Frozen Tissues Shows Distinct Compartments of Interlobular/Intralobular, Intra-Acinar, and Intra-Islet Fat Deposition","authors":"Claudio Luchini , Carlotta Franzina , Federico Caldart , Nicolò De Pretis , Manola Crestani , Massimo Donadelli , Paola Mattiolo , Alessandra Fiore , Federica Danzi , Riccardo De Robertis , Michele Bevere , Roberto Baldan , Laura Tommasi , Nicolò Vianini , Paolo Bernardi , Mirco Galiè , Antonio Pea , Rachele Ciccocioppo , Mirko D’Onofrio , Roberto Salvia , Luca Frulloni","doi":"10.1016/j.labinv.2025.104214","DOIUrl":"10.1016/j.labinv.2025.104214","url":null,"abstract":"<div><div>Obesity-related diseases and perturbations of fat metabolism represent some of the most common health challenges. In this complex scenario, recent evidence has suggested the emergence of a condition related to fat accumulation in the pancreas, which is generally referred to as fatty pancreas disease. This study aimed to clarify the different compartments of intrapancreatic fat deposition. The study cohort is represented by 100 patients who underwent pancreatic surgical resection. The pancreatic neck margin was analyzed with hematoxylin and eosin for evaluating tissue composition and with Oil Red O, a fat-specific histochemical staining, highlighting lipid droplets as red signals, for evaluating the presence of intracellular fat. Two cases were also analyzed with electron microscopy as cross-sectional validation. Regarding tissue composition, the most prevalent component was normal pancreatic parenchyma (mean value, 71.8%), followed by fibrosis (17.3%) and interlobular/intralobular fat (10.9%). Regarding intracellular fat deposition, Oil Red O–positive intracytoplasmic lipid droplets were present in most patients. The tissue areas with the highest levels of fat deposition were Langerhans’ islets, with neuroendocrine/insular cells showing more commonly a diffuse pattern of fat accumulation (>75% of cells). Electron microscopy confirmed the presence of intracytoplasmic lipid vacuoles in neuroendocrine/insular cells. Our findings showed the presence of different compartments of intrapancreatic fat deposition, both in terms of tissue composition and intracellular compartmentalization. Understanding the mechanisms of fat deposition in the pancreas is crucial toward improving the general knowledge on fatty pancreas disease, also opening new perspectives for the study of lipid metabolism and the treatment of fat-related diseases.</div></div>","PeriodicalId":17930,"journal":{"name":"Laboratory Investigation","volume":"105 11","pages":"Article 104214"},"PeriodicalIF":4.2,"publicationDate":"2025-08-19","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144959249","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2025-08-13DOI: 10.1016/j.labinv.2025.104225
Jaekwon Seok , Hee Jeong Kwak , Chan-Koo Kang , Ah Ram Kim , Woo Suk Choi , Hyoung Keun Park , Sung Hyun Paick , Hyeong Gon Kim , Yeonjoo Kwak , Tak-Il Jeon , Kyung Min Lim , Baeckseung Lee , Aram Kim , Ssang-Goo Cho
{"title":"Corrigendum to ‘Development of a Technique for Diagnosis and Screening of Superficial Bladder Cancer by Cell-Pellet DNA From Urine Sample’ [Laboratory Investigation 105 (2025) 104124]","authors":"Jaekwon Seok , Hee Jeong Kwak , Chan-Koo Kang , Ah Ram Kim , Woo Suk Choi , Hyoung Keun Park , Sung Hyun Paick , Hyeong Gon Kim , Yeonjoo Kwak , Tak-Il Jeon , Kyung Min Lim , Baeckseung Lee , Aram Kim , Ssang-Goo Cho","doi":"10.1016/j.labinv.2025.104225","DOIUrl":"10.1016/j.labinv.2025.104225","url":null,"abstract":"","PeriodicalId":17930,"journal":{"name":"Laboratory Investigation","volume":"105 9","pages":"Article 104225"},"PeriodicalIF":4.2,"publicationDate":"2025-08-13","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144829716","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2025-08-06DOI: 10.1016/j.labinv.2025.104227
Hao Lin , Jia-Jie Liang , Chen-Xi Zhang , Qi-Wen Man , Rui-Fang Li , Lin-Zhou Zhang , Bing Liu
Cellular senescence and its associated microenvironment play a pivotal role in tumor initiation and progression. Although ameloblastoma (AM) is classified as a benign tumor, it is characterized by local invasiveness and a high recurrence rate. In this study, we identified a distinct population of senescent epithelial cells using senescence-associated β-galactosidase staining and explored the role of this subpopulation in AM progression. The CellChat tool was utilized to map intercellular communication networks, revealing that this senescent cell cluster promotes stemness in neighboring cells and drives angiogenesis and osteoclastogenesis, which was subsequently confirmed by a series of in vitro experiments. Moreover, conditioned medium from these senescent cells significantly enhanced tumor growth in patient-derived organoids. Clinical data further demonstrated that elevated levels of cellular senescence were strongly associated with greater tumor invasiveness and poorer prognosis in AM patients. In conclusion, our findings suggest that targeting this specific subset of senescent epithelial cells may offer a novel therapeutic approach for AM management, potentially reducing tumor aggressiveness and recurrence.
{"title":"Senescent Epithelial Cells Serve as Invasive Growth Drivers in Ameloblastoma","authors":"Hao Lin , Jia-Jie Liang , Chen-Xi Zhang , Qi-Wen Man , Rui-Fang Li , Lin-Zhou Zhang , Bing Liu","doi":"10.1016/j.labinv.2025.104227","DOIUrl":"10.1016/j.labinv.2025.104227","url":null,"abstract":"<div><div>Cellular senescence and its associated microenvironment play a pivotal role in tumor initiation and progression. Although ameloblastoma (AM) is classified as a benign tumor, it is characterized by local invasiveness and a high recurrence rate. In this study, we identified a distinct population of senescent epithelial cells using senescence-associated β-galactosidase staining and explored the role of this subpopulation in AM progression. The CellChat tool was utilized to map intercellular communication networks, revealing that this senescent cell cluster promotes stemness in neighboring cells and drives angiogenesis and osteoclastogenesis, which was subsequently confirmed by a series of <em>in vitro</em> experiments. Moreover, conditioned medium from these senescent cells significantly enhanced tumor growth in patient-derived organoids. Clinical data further demonstrated that elevated levels of cellular senescence were strongly associated with greater tumor invasiveness and poorer prognosis in AM patients. In conclusion, our findings suggest that targeting this specific subset of senescent epithelial cells may offer a novel therapeutic approach for AM management, potentially reducing tumor aggressiveness and recurrence.</div></div>","PeriodicalId":17930,"journal":{"name":"Laboratory Investigation","volume":"105 11","pages":"Article 104227"},"PeriodicalIF":4.2,"publicationDate":"2025-08-06","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144804359","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
The role of poly (adenosine diphosphate [ADP]-ribose) polymerase 1 binding protein (PARPBP) in hepatocellular carcinoma (HCC) prognosis remains unclear. This study investigated PARPBP expression in HCC clinical samples and evaluated its clinicopathological significance in relation to CD8-positive tumor-infiltrating lymphocytes (CD8+ TILs). PARPBP expression and CD8+ TIL density were assessed in surgical specimens from 96 patients with HCC. We performed immunohistochemical analysis to determine PARPBP protein levels, which were correlated with clinicopathological features and patient outcomes. Additionally, we performed experiments using a doxycycline-inducible short hairpin RNA system targeting PARPBP in Huh7 cells. Results indicated that moderately and poorly differentiated HCC had significantly higher PARPBP expression than well-differentiated tumors. Patients with high PARPBP expression exhibited shorter recurrence-free survival and overall survival than those with low expression (both P < .001). Multivariate analysis showed that high PARPBP expression (hazard ratio [HR], 2.806; P = .002), high CD8+ TIL density (HR, 0.148; P < .001), and α-fetoprotein ≥ 10 ng/mL (HR, 1.904; P = .047) were independent predictors of prognosis. Combined analysis revealed that patients with high PARPBP expression and low CD8+ TIL density had the worst recurrence-free survival and overall survival outcomes (both P < .001). In vitro experiments showed that the mRNA expression of PARPBP was higher in liver cancer cell lines than in normal liver cells and that PARPBP knockdown suppressed huh7 cell proliferation. In conclusion, elevated PARPBP expression was associated with poor differentiation and survival in patients with HCC. Furthermore, the combination of high PARPBP expression and low CD8+ TIL density improved prognostic accuracy.
{"title":"Prognostic Significance of Poly (ADP-Ribose) Polymerase 1 Binding Protein Expression and CD8+ Tumor-Infiltrating Lymphocytes in Hepatocellular Carcinoma","authors":"Masako Okada , Misako Sato-Matsubara , Masaru Enomoto , Truong Huu Hoang , Sawako Uchida-Kobayashi , Akihiro Tamori , Tsutomu Matsubara , Kenichi Kohashi , Takeaki Ishizawa , Norifumi Kawada","doi":"10.1016/j.labinv.2025.104226","DOIUrl":"10.1016/j.labinv.2025.104226","url":null,"abstract":"<div><div>The role of poly (adenosine diphosphate [ADP]-ribose) polymerase 1 binding protein (PARPBP) in hepatocellular carcinoma (HCC) prognosis remains unclear. This study investigated PARPBP expression in HCC clinical samples and evaluated its clinicopathological significance in relation to CD8-positive tumor-infiltrating lymphocytes (CD8<sup>+</sup> TILs). PARPBP expression and CD8<sup>+</sup> TIL density were assessed in surgical specimens from 96 patients with HCC. We performed immunohistochemical analysis to determine PARPBP protein levels, which were correlated with clinicopathological features and patient outcomes. Additionally, we performed experiments using a doxycycline-inducible short hairpin RNA system targeting PARPBP in Huh7 cells. Results indicated that moderately and poorly differentiated HCC had significantly higher PARPBP expression than well-differentiated tumors. Patients with high PARPBP expression exhibited shorter recurrence-free survival and overall survival than those with low expression (both <em>P</em> < .001). Multivariate analysis showed that high PARPBP expression (hazard ratio [HR], 2.806; <em>P</em> = .002), high CD8<sup>+</sup> TIL density (HR, 0.148; <em>P</em> < .001), and α-fetoprotein ≥ 10 ng/mL (HR, 1.904; <em>P</em> = .047) were independent predictors of prognosis. Combined analysis revealed that patients with high PARPBP expression and low CD8<sup>+</sup> TIL density had the worst recurrence-free survival and overall survival outcomes (both <em>P</em> < .001). In vitro experiments showed that the mRNA expression of <em>PARPBP</em> was higher in liver cancer cell lines than in normal liver cells and that <em>PARPBP</em> knockdown suppressed huh7 cell proliferation. In conclusion, elevated PARPBP expression was associated with poor differentiation and survival in patients with HCC. Furthermore, the combination of high PARPBP expression and low CD8<sup>+</sup> TIL density improved prognostic accuracy.</div></div>","PeriodicalId":17930,"journal":{"name":"Laboratory Investigation","volume":"105 11","pages":"Article 104226"},"PeriodicalIF":4.2,"publicationDate":"2025-08-06","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144804358","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2025-07-29DOI: 10.1016/j.labinv.2025.104220
Mohamed Omar , Giuseppe Nicolo’ Fanelli , Fabio Socciarelli , Varun Ullanat , Sreekar Reddy Puchala , James Wen , Alex Chowdhury , Itzel Valencia , Cristian Scatena , Luigi Marchionni , Renato Umeton , Massimo Loda
Conventional histopathology has traditionally been the cornerstone of disease diagnosis, relying on qualitative or semiquantitative visual inspection of tissue sections to detect pathological changes. Singleplex immunohistochemistry (IHC), although effective in detecting specific biomarkers, is often limited by its single-marker focus, which constrains its ability to capture the complexity of the tissue environment. The introduction of multiplexed imaging technologies, such as multiplex IHC and multiplex immunofluorescence, has been transformative, enabling the simultaneous visualization of multiple biomarkers within a single tissue section. These approaches complement morphology with quantitative multimarker data and spatial context, providing a more comprehensive view of cellular interactions and disease mechanisms. However, the rich data from multiplex IHC/multiplex immunofluorescence experiments come with significant analytical challenges, as large multichannel images require comprehensive processing to transform raw imaging data into quantitative and meaningful information. This review focuses on the standard digital image analysis workflow for multiplex imaging in pathology, covering each step from image acquisition and preprocessing to cell segmentation and biomarker quantification. We discuss the common open-source tools that support each step to guide users in selecting appropriate solutions. By outlining an end-to-end pipeline with concrete examples, this review is intended for practicing pathologists and researchers with limited computational expertise. It provides practical guidance and best practices to help integrate multiplex image analysis into routine pathology workflows and translational research, bridging the gap between advanced imaging technology and day-to-day diagnostic practice.
{"title":"Antibody-Based Multiplex Image Analysis: Standard Analytical Workflows and Artificial Intelligence Tools for Pathologists","authors":"Mohamed Omar , Giuseppe Nicolo’ Fanelli , Fabio Socciarelli , Varun Ullanat , Sreekar Reddy Puchala , James Wen , Alex Chowdhury , Itzel Valencia , Cristian Scatena , Luigi Marchionni , Renato Umeton , Massimo Loda","doi":"10.1016/j.labinv.2025.104220","DOIUrl":"10.1016/j.labinv.2025.104220","url":null,"abstract":"<div><div>Conventional histopathology has traditionally been the cornerstone of disease diagnosis, relying on qualitative or semiquantitative visual inspection of tissue sections to detect pathological changes. Singleplex immunohistochemistry (IHC), although effective in detecting specific biomarkers, is often limited by its single-marker focus, which constrains its ability to capture the complexity of the tissue environment. The introduction of multiplexed imaging technologies, such as multiplex IHC and multiplex immunofluorescence, has been transformative, enabling the simultaneous visualization of multiple biomarkers within a single tissue section. These approaches complement morphology with quantitative multimarker data and spatial context, providing a more comprehensive view of cellular interactions and disease mechanisms. However, the rich data from multiplex IHC/multiplex immunofluorescence experiments come with significant analytical challenges, as large multichannel images require comprehensive processing to transform raw imaging data into quantitative and meaningful information. This review focuses on the standard digital image analysis workflow for multiplex imaging in pathology, covering each step from image acquisition and preprocessing to cell segmentation and biomarker quantification. We discuss the common open-source tools that support each step to guide users in selecting appropriate solutions. By outlining an end-to-end pipeline with concrete examples, this review is intended for practicing pathologists and researchers with limited computational expertise. It provides practical guidance and best practices to help integrate multiplex image analysis into routine pathology workflows and translational research, bridging the gap between advanced imaging technology and day-to-day diagnostic practice.</div></div>","PeriodicalId":17930,"journal":{"name":"Laboratory Investigation","volume":"105 10","pages":"Article 104220"},"PeriodicalIF":4.2,"publicationDate":"2025-07-29","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144760448","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2025-07-29DOI: 10.1016/j.labinv.2025.104221
Takeshi Iwasaki, Mari Masunaga, Takumi Tomonaga, Yosuke Masumoto, Yuri Akiyama, Yoshinao Oda
N6-methyladenosine (m6A), a widespread RNA modification, plays a vital role in various biological processes, including carcinogenesis, tumor progression, and immune regulation. We conducted this study to investigate the relationship between m6A regulators, such as METTL3, METTL14, WTAP, FTO, ALKBH5, and YTHDF1-3, and their association with c-Myc and programmed death ligand 1 (PD-L1) expression in leiomyosarcoma (LMS). The expression of these epitranscriptome regulator genes was evaluated using the next-generation sequencing data of 53 patients with LMS obtained from an online public database. We next determined the relationship between m6A regulators and c-Myc and CD274 (PD-L1) mRNA expression in an LMS cell line. We also performed immunohistochemical staining of 69 LMS cases. Immunohistochemical staining showed that cases with higher expression of METTL3, METTL14, ALKBH5, FTO, and WTAP exhibited higher c-Myc expression, and cases with higher expression of ALKBH5, YTHDF2, and WTAP exhibited higher mitotic activity. Gene set enrichment analysis revealed that METTL3, METTL14, and FTO knockdown significantly suppressed c-Myc target gene expression. Knockdown of METTL3, ALKBH5, YTHDF1, WTAP, and FTO in LMS cell lines reduced cell proliferation. These results suggest the relationship between m6A modifications and c-Myc-driven oncogenesis. Moreover, knockdown of YTHDF2 inhibited interferon gamma–induced PD-L1 expression, suggesting its role in immune evasion through PD-L1 regulation. Multivariate Cox proportional hazards analysis revealed that lower YTHDF2 expression and higher WTAP expression are unfavorable prognostic factors. These findings provide potential therapeutic targets for LMS, particularly in combination with immune checkpoint inhibitors. Further investigation into the molecular mechanisms of m6A-mediated regulation of PD-L1 and c-Myc expression is required to develop more effective treatments for LMS.
{"title":"Comprehensive Analysis of N6-Methyladenosine (m6A) RNA Methylation Regulators in Soft Tissue Leiomyosarcoma","authors":"Takeshi Iwasaki, Mari Masunaga, Takumi Tomonaga, Yosuke Masumoto, Yuri Akiyama, Yoshinao Oda","doi":"10.1016/j.labinv.2025.104221","DOIUrl":"10.1016/j.labinv.2025.104221","url":null,"abstract":"<div><div>N6-methyladenosine (m6A), a widespread RNA modification, plays a vital role in various biological processes, including carcinogenesis, tumor progression, and immune regulation. We conducted this study to investigate the relationship between m6A regulators, such as METTL3, METTL14, WTAP, FTO, ALKBH5, and YTHDF1-3, and their association with c-Myc and programmed death ligand 1 (PD-L1) expression in leiomyosarcoma (LMS). The expression of these epitranscriptome regulator genes was evaluated using the next-generation sequencing data of 53 patients with LMS obtained from an online public database. We next determined the relationship between m6A regulators and <em>c-Myc</em> and <em>CD274 (PD-L1)</em> mRNA expression in an LMS cell line. We also performed immunohistochemical staining of 69 LMS cases. Immunohistochemical staining showed that cases with higher expression of METTL3, METTL14, ALKBH5, FTO, and WTAP exhibited higher c-Myc expression, and cases with higher expression of ALKBH5, YTHDF2, and WTAP exhibited higher mitotic activity. Gene set enrichment analysis revealed that <em>METTL3</em>, <em>METTL14</em>, and <em>FTO</em> knockdown significantly suppressed c-Myc target gene expression. Knockdown of <em>METTL3</em>, <em>ALKBH5</em>, <em>YTHDF1</em>, <em>WTAP</em>, and <em>FTO</em> in LMS cell lines reduced cell proliferation. These results suggest the relationship between m6A modifications and c-Myc-driven oncogenesis. Moreover, knockdown of <em>YTHDF2</em> inhibited interferon gamma–induced <em>PD-L1</em> expression, suggesting its role in immune evasion through PD-L1 regulation. Multivariate Cox proportional hazards analysis revealed that lower YTHDF2 expression and higher WTAP expression are unfavorable prognostic factors. These findings provide potential therapeutic targets for LMS, particularly in combination with immune checkpoint inhibitors. Further investigation into the molecular mechanisms of m6A-mediated regulation of PD-L1 and c-Myc expression is required to develop more effective treatments for LMS.</div></div>","PeriodicalId":17930,"journal":{"name":"Laboratory Investigation","volume":"105 11","pages":"Article 104221"},"PeriodicalIF":4.2,"publicationDate":"2025-07-29","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144760449","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
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