This review provides a personal overview of significant scientific developments in the thrombospondin field during the course of my career. Thrombospondins are multidomain, multimeric, calcium-binding extracellular glycoproteins with context-specific roles in tissue organisation. They act at cell surfaces and within ECM to regulate cell phenotype and signalling, differentiation and assembly of collagenous ECM, along with tissue-specific roles in cartilage, angiogenesis and synaptic function. More recently, intracellular, homeostatic roles have also been identified. Resolution of structures for the major domains of mammalian thrombospondins has facilitated major advances in understanding thrombospondin biology from molecule to tissue; for example, in illuminating molecular consequences of disease-causing coding mutations in human pseudoachrondroplasia. Although principally studied in vertebrates, thrombospondins are amongst the most ancient of animal ECM proteins, with many invertebrates encoding a single thrombospondin and the thrombospondin gene family of vertebrates originating through gene duplications. Moreover, thrombospondins form one branch of a thrombospondin superfamily that debuted at the origin of metazoans. The super-family includes additional sub-groups, present only in invertebrates, that differ in N-terminal domain organisation, share the distinctive TSP C-terminal region domain architecture and, to the limited extent studied to date, apparently contribute to tissue development and organisation. Finally, major lines of translational research are discussed, related to fibrosis; TSP1, TSP2 and inhibition of angiogenesis; and the alleviation of chronic cartilage tissue pathologies in pseudoachrondroplasia.
这篇综述对我职业生涯中凝血酶原领域的重大科学进展进行了个人概述。血栓软蛋白是多域、多聚体、钙结合细胞外糖蛋白,在组织结构中具有特定的作用。它们在细胞表面和 ECM 内发挥作用,调节细胞表型和信号、分化和胶原 ECM 的组装,并在软骨、血管生成和突触功能中发挥组织特异性作用。最近,人们还发现了细胞内的平衡作用。哺乳动物血栓软骨素主要结构域结构的解析促进了从分子到组织对血栓软骨素生物学认识的重大进展;例如,阐明了人类假性软骨软化症中致病编码突变的分子后果。虽然主要是在脊椎动物中进行研究,但血栓软蛋白是最古老的动物 ECM 蛋白之一,许多无脊椎动物编码单一的血栓软蛋白,而脊椎动物的血栓软蛋白基因家族则是通过基因复制形成的。此外,血栓软蛋白构成了血栓软蛋白超家族的一个分支,该超家族在元古宙起源时就已出现。该超家族包括仅存在于无脊椎动物中的其他亚群,这些亚群在 N 端结构域的组织结构上有所不同,但共享独特的 TSP C 端区域结构域结构,并且在迄今为止有限的研究范围内,显然有助于组织的发育和组织。最后,讨论了与纤维化、TSP1、TSP2 和血管生成抑制以及减轻假性软骨增生症中慢性软骨组织病变有关的主要转化研究方向。
{"title":"Thrombospondins: Conserved mediators and modulators of metazoan extracellular matrix","authors":"Josephine C. Adams","doi":"10.1111/iep.12517","DOIUrl":"10.1111/iep.12517","url":null,"abstract":"<p>This review provides a personal overview of significant scientific developments in the thrombospondin field during the course of my career. Thrombospondins are multidomain, multimeric, calcium-binding extracellular glycoproteins with context-specific roles in tissue organisation. They act at cell surfaces and within ECM to regulate cell phenotype and signalling, differentiation and assembly of collagenous ECM, along with tissue-specific roles in cartilage, angiogenesis and synaptic function. More recently, intracellular, homeostatic roles have also been identified. Resolution of structures for the major domains of mammalian thrombospondins has facilitated major advances in understanding thrombospondin biology from molecule to tissue; for example, in illuminating molecular consequences of disease-causing coding mutations in human pseudoachrondroplasia. Although principally studied in vertebrates, thrombospondins are amongst the most ancient of animal ECM proteins, with many invertebrates encoding a single thrombospondin and the thrombospondin gene family of vertebrates originating through gene duplications. Moreover, thrombospondins form one branch of a thrombospondin superfamily that debuted at the origin of metazoans. The super-family includes additional sub-groups, present only in invertebrates, that differ in N-terminal domain organisation, share the distinctive TSP C-terminal region domain architecture and, to the limited extent studied to date, apparently contribute to tissue development and organisation. Finally, major lines of translational research are discussed, related to fibrosis; TSP1, TSP2 and inhibition of angiogenesis; and the alleviation of chronic cartilage tissue pathologies in pseudoachrondroplasia.</p>","PeriodicalId":14157,"journal":{"name":"International Journal of Experimental Pathology","volume":null,"pages":null},"PeriodicalIF":1.8,"publicationDate":"2024-09-12","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://onlinelibrary.wiley.com/doi/epdf/10.1111/iep.12517","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142255575","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Matrix metalloproteinase (MMP)-12 has been reported to have diverse functions, including regulation of immune reactions and anti-inflammatory effects, but the potential roles of MMP-12 in kidney injury have not been fully elucidated. This study aimed to determine whether MMP-12 contributes to tubulointerstitial injury in a unilateral ureteric obstruction (UUO) model. MMP-12-deficient (MMP-12−/−) mice and C57BL/6J mice as controls (MMP-12+/+) were subjected to UUO and analysed 7 days after UUO. To analyse the functions of MMP-12 on monocytes/macrophages, we generated MMP-12-deficient, irradiated, chimeric mice (BM-MMP-12−/−) and performed UUO. Bone marrow-derived macrophages (BMDMs) were isolated from both groups of mice and used for investigations. MMP-12−/− mice showed exacerbation of macrophage accumulation and interstitial fibrosis in the UUO-kidney compared with control mice. BM-MMP-12−/− mice also showed exacerbation of kidney injury. UUO induced accumulation of Ly6C+ macrophages in MMP-12−/− mice compared with control mice. Increases in inflammatory cytokine (tumour necrosis factor α, interleukin [IL]-1β, IL-6) levels from BMDMs after lipopolysaccharide stimulation were higher in MMP-12−/− mice than in MMP-12+/+ mice. MMP-12 may play protective roles against kidney injury by UUO in mice, decreasing inflammatory cytokines from BMDMs and macrophage accumulation.
{"title":"Renal protective roles of macrophage matrix metalloproteinase-12 in mice with obstructed kidneys","authors":"Shunichiro Hanai, Daiki Nakagomi, Kotaro Suzuki, Hiroshi Nakajima, Fumihiko Furuya","doi":"10.1111/iep.12516","DOIUrl":"10.1111/iep.12516","url":null,"abstract":"<p>Matrix metalloproteinase (MMP)-12 has been reported to have diverse functions, including regulation of immune reactions and anti-inflammatory effects, but the potential roles of MMP-12 in kidney injury have not been fully elucidated. This study aimed to determine whether MMP-12 contributes to tubulointerstitial injury in a unilateral ureteric obstruction (UUO) model. MMP-12-deficient (MMP-12<sup>−/−</sup>) mice and C57BL/6J mice as controls (MMP-12<sup>+/+</sup>) were subjected to UUO and analysed 7 days after UUO. To analyse the functions of MMP-12 on monocytes/macrophages, we generated MMP-12-deficient, irradiated, chimeric mice (BM-MMP-12<sup>−/−</sup>) and performed UUO. Bone marrow-derived macrophages (BMDMs) were isolated from both groups of mice and used for investigations. MMP-12<sup>−/−</sup> mice showed exacerbation of macrophage accumulation and interstitial fibrosis in the UUO-kidney compared with control mice. BM-MMP-12<sup>−/−</sup> mice also showed exacerbation of kidney injury. UUO induced accumulation of Ly6C<sup>+</sup> macrophages in MMP-12<sup>−/−</sup> mice compared with control mice. Increases in inflammatory cytokine (tumour necrosis factor α, interleukin [IL]-1β, IL-6) levels from BMDMs after lipopolysaccharide stimulation were higher in MMP-12<sup>−/−</sup> mice than in MMP-12<sup>+/+</sup> mice. MMP-12 may play protective roles against kidney injury by UUO in mice, decreasing inflammatory cytokines from BMDMs and macrophage accumulation.</p>","PeriodicalId":14157,"journal":{"name":"International Journal of Experimental Pathology","volume":null,"pages":null},"PeriodicalIF":1.8,"publicationDate":"2024-08-20","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142008777","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Zinc levels in breast cancer tissues have been reported to be higher than those in normal tissues. In addition, the expression levels of zinc transporters, including ZnT5 and ZnT6, are reportedly higher in breast cancer than in normal breast tissues. ZnT5 and ZnT6 also contribute to heterodimer formation and are involved in several biological functions. However, the functions of ZnT5 and ZnT6 heterodimers in breast cancer remain unknown. Therefore, we first investigated the immunolocalization of ZnT5 and ZnT6 in pathological breast cancer specimens and in MCF-7 and T-47D breast cancer cells. Next, we used small interfering RNA to assess cell viability and migration in ZnT5 knockdown MCF-7 and T-47D cells. Immunohistochemical analysis showed that the number of ZnT5-positive breast cancer cells was inversely correlated with the pathologic N factor status. ZnT5 knockdown had no effect on cell viability in the presence of 100 μM ZnCl2 in MCF-7 and T-47D cells. In a wound healing assay, 100 μM ZnCl2 treatment inhibited cell migration of MCF-7 and T-47D cells, whereas ZnT5 knockdown promoted cell migration, decreased E-cadherin expression and increased vimentin, slug and matrix metalloproteinase 9 expression. Antibody arrays showed that ZnT5 knockdown increased the expression of SMAD1, and that dorsomorphin treatment inhibited the promotion of migratory ability induced by ZnT5 knockdown. The results of this study revealed that both ZnT5 may be involved in less aggressive breast cancer subtypes, possibly through inhibition of cell migration.
{"title":"Zinc transporter ZnT5 is associated with epithelial mesenchymal transition via SMAD1 in breast cancer","authors":"Erina Iwabuchi, Yasuhiro Miki, Junyao Xu, Ayako Kanai, Takanori Ishida, Hironobu Sasano, Takashi Suzuki","doi":"10.1111/iep.12515","DOIUrl":"10.1111/iep.12515","url":null,"abstract":"<p>Zinc levels in breast cancer tissues have been reported to be higher than those in normal tissues. In addition, the expression levels of zinc transporters, including ZnT5 and ZnT6, are reportedly higher in breast cancer than in normal breast tissues. ZnT5 and ZnT6 also contribute to heterodimer formation and are involved in several biological functions. However, the functions of ZnT5 and ZnT6 heterodimers in breast cancer remain unknown. Therefore, we first investigated the immunolocalization of ZnT5 and ZnT6 in pathological breast cancer specimens and in MCF-7 and T-47D breast cancer cells. Next, we used small interfering RNA to assess cell viability and migration in ZnT5 knockdown MCF-7 and T-47D cells. Immunohistochemical analysis showed that the number of ZnT5-positive breast cancer cells was inversely correlated with the pathologic N factor status. ZnT5 knockdown had no effect on cell viability in the presence of 100 μM ZnCl<sub>2</sub> in MCF-7 and T-47D cells. In a wound healing assay, 100 μM ZnCl<sub>2</sub> treatment inhibited cell migration of MCF-7 and T-47D cells, whereas ZnT5 knockdown promoted cell migration, decreased E-cadherin expression and increased vimentin, slug and matrix metalloproteinase 9 expression. Antibody arrays showed that ZnT5 knockdown increased the expression of SMAD1, and that dorsomorphin treatment inhibited the promotion of migratory ability induced by ZnT5 knockdown. The results of this study revealed that both ZnT5 may be involved in less aggressive breast cancer subtypes, possibly through inhibition of cell migration.</p>","PeriodicalId":14157,"journal":{"name":"International Journal of Experimental Pathology","volume":null,"pages":null},"PeriodicalIF":1.8,"publicationDate":"2024-08-13","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://onlinelibrary.wiley.com/doi/epdf/10.1111/iep.12515","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"141975585","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Feline primary hypertrophic cardiomyopathy (HCM) is an intrinsic myocardial disease characterized by concentric hypertrophy of the left ventricle. In the present study, we investigated the microRNA-mRNA regulatory network in feline myocardial tissue affected by primary (HCMI) and secondary HCM (HCMII). MRNA expression levels of sarcomeric genes, including, TNNT2, TNNI3, MYH7, MYBPC3, TPM1 and ACTC1 were assessed in the FFPE myocardial tissues. FFPE tissues from healthy cats were sequenced by the NGS, to explore, in the entire non-deposited miRNome, the expression level of microRNAs targeting the complementary sequences of selected sarcomeric mRNAs. The sarcomeric genes TNNT2, MYH7, MYBPC3 and TPM1 showed a statistically significant upregulation in HCMI compared to HCMII (p < .01), except ACTC1 which was downregulated (p < .01); TNNI3 showed no statistically significant difference. In HCMII miR-122-5p, miR-338-3p, miR-484, miR-370-3p, miR-92b-3p, miR-375 and miR-370-3p showed a significant upregulation (p < .01) compared to control. The exception was miR-30a-5p which showed downregulation. Worthy of note is the 4-fold higher expression of miR-370-3p, a key regulator of MYBPC3, in HMCI compared to HMCII. This research does not solve the aetiological mystery of HCM, but it may help to find a way to help diagnose and define the prognosis of HCM in cats.
{"title":"Feline hypertrophic cardiomyopathy: Does the microRNA-mRNA regulatory network contribute to heart sarcomeric protein remodelling?","authors":"Gabriella Guelfi, Noemi Venanzi, Camilla Capaccia, Valentina Stefanetti, Chiara Brachelente, Monica Sforna, Francesco Porciello, Elvio Lepri","doi":"10.1111/iep.12514","DOIUrl":"10.1111/iep.12514","url":null,"abstract":"<p>Feline primary hypertrophic cardiomyopathy (HCM) is an intrinsic myocardial disease characterized by concentric hypertrophy of the left ventricle. In the present study, we investigated the microRNA-mRNA regulatory network in feline myocardial tissue affected by primary (HCMI) and secondary HCM (HCMII). MRNA expression levels of sarcomeric genes, including, <i>TNNT2</i>, <i>TNNI3</i>, <i>MYH7</i>, <i>MYBPC3</i>, <i>TPM1</i> and <i>ACTC1</i> were assessed in the FFPE myocardial tissues. FFPE tissues from healthy cats were sequenced by the NGS, to explore, in the entire non-deposited miRNome, the expression level of microRNAs targeting the complementary sequences of selected sarcomeric mRNAs. The sarcomeric genes <i>TNNT2</i>, <i>MYH7</i>, <i>MYBPC3</i> and <i>TPM1</i> showed a statistically significant upregulation in HCMI compared to HCMII (<i>p</i> < .01), except <i>ACTC1</i> which was downregulated (<i>p</i> < .01); <i>TNNI3</i> showed no statistically significant difference. In HCMII miR-122-5p, miR-338-3p, miR-484, miR-370-3p, miR-92b-3p, miR-375 and miR-370-3p showed a significant upregulation (<i>p</i> < .01) compared to control. The exception was miR-30a-5p which showed downregulation. Worthy of note is the 4-fold higher expression of miR-370-3p, a key regulator of <i>MYBPC3</i>, in HMCI compared to HMCII. This research does not solve the aetiological mystery of HCM, but it may help to find a way to help diagnose and define the prognosis of HCM in cats.</p>","PeriodicalId":14157,"journal":{"name":"International Journal of Experimental Pathology","volume":null,"pages":null},"PeriodicalIF":1.8,"publicationDate":"2024-08-13","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://onlinelibrary.wiley.com/doi/epdf/10.1111/iep.12514","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"141975584","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
{"title":"British Society for Matrix Biology Spring Meeting 2024: The Dynamic matrix—Mechanics, Ageing and Repair","authors":"","doi":"10.1111/iep.12512","DOIUrl":"10.1111/iep.12512","url":null,"abstract":"","PeriodicalId":14157,"journal":{"name":"International Journal of Experimental Pathology","volume":null,"pages":null},"PeriodicalIF":1.8,"publicationDate":"2024-07-23","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"141748155","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Oesophageal cancer (EC) is a malignancy which accounts for a substantial number of cancer-related deaths worldwide. The molecular mechanisms underlying the pathogenesis of EC have not been fully elucidated. GSE17351 and GSE20347 data sets from the Gene Expression Omnibus (GEO) database were employed to screen differentially expressed genes (DEGs). Reverse transcription quantitative PCR (RT-qPCR) was used to examine hub gene expression. ECA-109 and TE-12 cells were transfected using the pcDNA3.1 expression vector encoding GABRP. The cell counting kit-8 (CCK-8), cell scratch and Transwell assays were performed to assess the effect of GABRP on EC cell proliferation, migration and invasion. Epithelial-mesenchymal transition (EMT)-associated protein levels were measured by Western blotting. Subsequently, CFTR was knocked down to verify whether GABRP affected biological events in EC cells by targeting CFTR. Seven hub genes were identified, including GABRP, FLG, ENAH, KLF4, CD24, ABLIM3 and ABLIM1, which all could be used as diagnostic biomarkers for EC. The RT-qPCR results indicated that the expression levels of GABRP, FLG, KLF4, CD24, ABLIM3 and ABLIM1 were downregulated, whereas the expression level of ENAH was upregulated. In vitro functional assays demonstrated that GABRP overexpression suppressed the proliferation, migration, invasion and EMT of EC cells. Mechanistically, GABRP promoted the expression of CFTR, and CFTR knockdown significantly counteracted the influence of GABRP overexpression on biological events in EC cells. Overexpression of GABRP inhibited EC progression by increasing CFTR expression, which might be a new target for EC treatment.
{"title":"GABRP inhibits the progression of oesophageal cancer by regulating CFTR: Integrating bioinformatics analysis and experimental validation","authors":"Jingzhi Zhang, Xue Liu, Ling Zeng, Ying Hu","doi":"10.1111/iep.12513","DOIUrl":"10.1111/iep.12513","url":null,"abstract":"<p>Oesophageal cancer (EC) is a malignancy which accounts for a substantial number of cancer-related deaths worldwide. The molecular mechanisms underlying the pathogenesis of EC have not been fully elucidated. GSE17351 and GSE20347 data sets from the Gene Expression Omnibus (GEO) database were employed to screen differentially expressed genes (DEGs). Reverse transcription quantitative PCR (RT-qPCR) was used to examine hub gene expression. ECA-109 and TE-12 cells were transfected using the pcDNA3.1 expression vector encoding <i>GABRP</i>. The cell counting kit-8 (CCK-8), cell scratch and Transwell assays were performed to assess the effect of <i>GABRP</i> on EC cell proliferation, migration and invasion. Epithelial-mesenchymal transition (EMT)-associated protein levels were measured by Western blotting. Subsequently, <i>CFTR</i> was knocked down to verify whether <i>GABRP</i> affected biological events in EC cells by targeting <i>CFTR</i>. Seven hub genes were identified, including <i>GABRP</i>, <i>FLG</i>, <i>ENAH</i>, <i>KLF4</i>, <i>CD24</i>, <i>ABLIM3</i> and <i>ABLIM1</i>, which all could be used as diagnostic biomarkers for EC. The RT-qPCR results indicated that the expression levels of <i>GABRP</i>, <i>FLG</i>, <i>KLF4</i>, <i>CD24</i>, <i>ABLIM3</i> and <i>ABLIM1</i> were downregulated, whereas the expression level of <i>ENAH</i> was upregulated. In vitro functional assays demonstrated that <i>GABRP</i> overexpression suppressed the proliferation, migration, invasion and EMT of EC cells. Mechanistically, <i>GABRP</i> promoted the expression of <i>CFTR</i>, and <i>CFTR</i> knockdown significantly counteracted the influence of <i>GABRP</i> overexpression on biological events in EC cells. Overexpression of <i>GABRP</i> inhibited EC progression by increasing <i>CFTR</i> expression, which might be a new target for EC treatment.</p>","PeriodicalId":14157,"journal":{"name":"International Journal of Experimental Pathology","volume":null,"pages":null},"PeriodicalIF":1.8,"publicationDate":"2024-07-11","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"141579639","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Papaioannou I, Owen JS and Yáñez-Muñoz RJ. Clinical applications of gene therapy for rare diseases: A review. Int J Exp Pathol 2023 Aug;104(4):154–176. doi: 10.1111/iep.12478. Epub 2023 May 13.
It has been brought to our attention that in paragraph 1 of section 5.3, some details regarding the design of Zolgensma were not accurate. Therefore, the following text was incorrect: “Zolgensma (Onasemnogene Abeparvovec)196–200 is an AAV-based gene supplementation treatment aimed at directly and permanently restoring SMN1 expression with a single dose. The design of the Zolgensma expression cassette is similar to Luxturna (Figure 5), in using the hybrid CMV–Chicken beta actin promoter to drive the expression of SMN1 cDNA. To enhance expression, the design incorporates an artificial intron (from SV40) and codon optimization. The sequence of AVXS-101 (the vector for Zolgensma) is proprietary and the exact optimizations are not in the public domain, but the effectiveness of this approach was documented by using a similar AAV9 platform.201–203 A self-complementary design (Figure 12) was employed, where one of the flanking ITRs was a specially engineered variant to synthesize genome dimers, rather than monomers.204”
This should have read: “Zolgensma (Onasemnogene Abeparvovec)196–200 is an AAV-based gene supplementation treatment aimed at directly and permanently restoring SMN protein production with a single dose. The design of the Zolgensma expression cassette is similar to Luxturna (Figure 5) in using the hybrid CMV–Chicken beta-actin promoter to drive the expression of SMN2 cDNA, and it includes the bovine growth hormone polyadenylation sequence. To enhance expression, the design incorporates an artificial intron (from SV40). Unusually, the sequence of the SMN2 cDNA in Zolgensma was not codon-optimized. The effectiveness of AVXS-101 (the vector for Zolgensma) was corroborated by the work of several other groups using similar self-complementary AAV9-SMN platforms201–203 (Figure 12), where one of the flanking ITRs was a specially engineered variant to synthesize genome dimers, rather than monomers.204”
{"title":"Correction to “Clinical applications of gene therapy for rare diseases: A review”","authors":"","doi":"10.1111/iep.12505","DOIUrl":"10.1111/iep.12505","url":null,"abstract":"<p>Papaioannou I, Owen JS and Yáñez-Muñoz RJ. Clinical applications of gene therapy for rare diseases: A review. <i>Int J Exp Pathol</i> 2023 Aug;104(4):154–176. doi: 10.1111/iep.12478. Epub 2023 May 13.</p><p>It has been brought to our attention that in paragraph 1 of section 5.3, some details regarding the design of Zolgensma were not accurate. Therefore, the following text was incorrect: “Zolgensma (Onasemnogene Abeparvovec)<sup>196–200</sup> is an AAV-based gene supplementation treatment aimed at directly and permanently restoring <i>SMN1</i> expression with a single dose. The design of the Zolgensma expression cassette is similar to Luxturna (Figure 5), in using the hybrid CMV–Chicken beta actin promoter to drive the expression of <i>SMN1</i> cDNA. To enhance expression, the design incorporates an artificial intron (from SV40) and codon optimization. The sequence of AVXS-101 (the vector for Zolgensma) is proprietary and the exact optimizations are not in the public domain, but the effectiveness of this approach was documented by using a similar AAV9 platform.<sup>201–203</sup> A self-complementary design (Figure 12) was employed, where one of the flanking ITRs was a specially engineered variant to synthesize genome dimers, rather than monomers.<sup>204</sup>”</p><p>This should have read: “Zolgensma (Onasemnogene Abeparvovec)<sup>196–200</sup> is an AAV-based gene supplementation treatment aimed at directly and permanently restoring SMN protein production with a single dose. The design of the Zolgensma expression cassette is similar to Luxturna (Figure 5) in using the hybrid CMV–Chicken beta-actin promoter to drive the expression of <i>SMN2</i> cDNA, and it includes the bovine growth hormone polyadenylation sequence. To enhance expression, the design incorporates an artificial intron (from SV40). Unusually, the sequence of the <i>SMN2</i> cDNA in Zolgensma was not codon-optimized. The effectiveness of AVXS-101 (the vector for Zolgensma) was corroborated by the work of several other groups using similar self-complementary AAV9-<i>SMN</i> platforms<sup>201–203</sup> (Figure 12), where one of the flanking ITRs was a specially engineered variant to synthesize genome dimers, rather than monomers.<sup>204</sup>”</p><p>We apologize for this error.</p>","PeriodicalId":14157,"journal":{"name":"International Journal of Experimental Pathology","volume":null,"pages":null},"PeriodicalIF":3.0,"publicationDate":"2024-05-15","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://onlinelibrary.wiley.com/doi/epdf/10.1111/iep.12505","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"140922113","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Larisse Longo, Bárbara Jonson Bartikoski, Valessa Emanoele Gabriel de Souza, Fernando Salvati, Carolina Uribe-Cruz, Guido Lenz, Ricardo Machado Xavier, Mário Reis Álvares-da-Silva, Eduardo Cremonese Filippi-Chiela
Morphometry of striated muscle fibres is critical for monitoring muscle health and function. Here, we evaluated functional parameters of skeletal and cardiac striated muscle in two experimental models using the Morphometric Analysis of Muscle Fibre tool (MusMA). The collagen-induced arthritis model was used to evaluate the function of skeletal striated muscle and the non-alcoholic fatty liver disease model was used for cardiac striated muscle analysis. After euthanasia, we used haeamatoxylin and eosin stained sections of skeletal and cardiac muscle to perform muscle fibre segmentation and morphometric analysis. Morphometric analysis classified muscle fibres into six subpopulations: normal, regular hypertrophic, irregular hypertrophic, irregular, irregular atrophic and regular atrophic. The percentage of atrophic fibres was associated with lower walking speed (p = 0.009) and lower body weight (p = 0.026), respectively. Fibres categorized as normal were associated with maximum grip strength (p < 0.001) and higher march speed (p < 0.001). In the evaluation of cardiac striated muscle fibres, the percentage of normal cardiomyocytes negatively correlated with cardiovascular risk markers such as the presence of abdominal adipose tissue (p = .003), miR-33a expression (p = .001) and the expression of miR-126 (p = .042) Furthermore, the percentage of atrophic cardiomyocytes correlated significantly with the Castelli risk index II (p = .014). MusMA is a simple and objective tool that allows the screening of striated muscle fibre morphometry, which can complement the diagnosis of muscle diseases while providing functional and prognostic information in basic and clinical research.
{"title":"Muscle fibre morphometric analysis (MusMA) correlates with muscle function and cardiovascular risk prognosis","authors":"Larisse Longo, Bárbara Jonson Bartikoski, Valessa Emanoele Gabriel de Souza, Fernando Salvati, Carolina Uribe-Cruz, Guido Lenz, Ricardo Machado Xavier, Mário Reis Álvares-da-Silva, Eduardo Cremonese Filippi-Chiela","doi":"10.1111/iep.12504","DOIUrl":"10.1111/iep.12504","url":null,"abstract":"<p>Morphometry of striated muscle fibres is critical for monitoring muscle health and function. Here, we evaluated functional parameters of skeletal and cardiac striated muscle in two experimental models using the Morphometric Analysis of Muscle Fibre tool (MusMA). The collagen-induced arthritis model was used to evaluate the function of skeletal striated muscle and the non-alcoholic fatty liver disease model was used for cardiac striated muscle analysis. After euthanasia, we used haeamatoxylin and eosin stained sections of skeletal and cardiac muscle to perform muscle fibre segmentation and morphometric analysis. Morphometric analysis classified muscle fibres into six subpopulations: normal, regular hypertrophic, irregular hypertrophic, irregular, irregular atrophic and regular atrophic. The percentage of atrophic fibres was associated with lower walking speed (<i>p</i> = 0.009) and lower body weight (<i>p</i> = 0.026), respectively. Fibres categorized as normal were associated with maximum grip strength (<i>p</i> < 0.001) and higher march speed (<i>p</i> < 0.001). In the evaluation of cardiac striated muscle fibres, the percentage of normal cardiomyocytes negatively correlated with cardiovascular risk markers such as the presence of abdominal adipose tissue (<i>p</i> = .003), miR-33a expression (<i>p</i> = .001) and the expression of miR-126 (<i>p</i> = .042) Furthermore, the percentage of atrophic cardiomyocytes correlated significantly with the Castelli risk index II (<i>p</i> = .014). MusMA is a simple and objective tool that allows the screening of striated muscle fibre morphometry, which can complement the diagnosis of muscle diseases while providing functional and prognostic information in basic and clinical research.</p>","PeriodicalId":14157,"journal":{"name":"International Journal of Experimental Pathology","volume":null,"pages":null},"PeriodicalIF":3.0,"publicationDate":"2024-05-09","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"140897872","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Management of lung cancer today obligates a mutational analysis of the epidermal growth factor receptor (EGFR) gene particularly when Tyrosine Kinase Inhibitor (TKI) therapy is being considered as part of prognostic stratification. This study evaluates the performance of automated microfluidics-based EGFR mutation detection and its significance in clinical diagnostic settings. Formalin-fixed, paraffin-embedded (FFPE) samples from NSCLC patients (n = 174) were included in a two-phase study. Phase I: Validation of the platform by comparing the results with conventional real-time PCR and next-generation sequencing (NGS) platform. Phase II: EGFR mutation detection on microfluidics-based platform as part of routine diagnostics workup. The microfluidics-based platform demonstrates 96.5% and 89.2% concordance with conventional real-time PCR and NGS, respectively. The system efficiently detects mutations across the EGFR gene with 88.23% sensitivity and 100% specificity. Out of 144 samples analysed in phase II, the platform generated valid results in 94% with mutation detected in 41% of samples. This microfluidics-based platform can detect as low as 5% mutant allele fractions from the FFPE samples. Therefore the microfluidics-based platform is a rapid, complete walkaway, with minimum tissue requirement (two sections of 5 μ thickness) and technical skill requirement. The method can detect clinically actionable EGFR mutations efficiently and can be considered a reliable diagnostic platform in resource-limited settings. From receiving samples to reporting the results this platform provides accurate data without much manual intervention. The study helped to devise an algorithm that emphasizes effective screening of the NSCLC cases for EGFR mutations with varying tumour content. Thus it helps in triaging the cases judiciously before proceeding with multigene testing.
{"title":"Microfluidics-based EGFR mutation detection and its implication in the resource-limited clinical setting","authors":"Pradnya Joshi, Prachi Gogte, Trupti Pai, Mamta Gurav, Dipika Dhanawade, Nupur Karnik, Gauri Deshpande, Rajiv Kaushal, Omshree Shetty","doi":"10.1111/iep.12503","DOIUrl":"10.1111/iep.12503","url":null,"abstract":"<p>Management of lung cancer today obligates a mutational analysis of the epidermal growth factor receptor (<i>EGFR)</i> gene particularly when Tyrosine Kinase Inhibitor (TKI) therapy is being considered as part of prognostic stratification. This study evaluates the performance of automated microfluidics-based <i>EGFR</i> mutation detection and its significance in clinical diagnostic settings. Formalin-fixed, paraffin-embedded (FFPE) samples from NSCLC patients (<i>n</i> = 174) were included in a two-phase study. Phase I: Validation of the platform by comparing the results with conventional real-time PCR and next-generation sequencing (NGS) platform. Phase II: <i>EGFR</i> mutation detection on microfluidics-based platform as part of routine diagnostics workup. The microfluidics-based platform demonstrates 96.5% and 89.2% concordance with conventional real-time PCR and NGS, respectively. The system efficiently detects mutations across the <i>EGFR</i> gene with 88.23% sensitivity and 100% specificity. Out of 144 samples analysed in phase II, the platform generated valid results in 94% with mutation detected in 41% of samples. This microfluidics-based platform can detect as low as 5% mutant allele fractions from the FFPE samples. Therefore the microfluidics-based platform is a rapid, complete walkaway, with minimum tissue requirement (two sections of 5 μ thickness) and technical skill requirement. The method can detect clinically actionable <i>EGFR</i> mutations efficiently and can be considered a reliable diagnostic platform in resource-limited settings. From receiving samples to reporting the results this platform provides accurate data without much manual intervention. The study helped to devise an algorithm that emphasizes effective screening of the NSCLC cases for <i>EGFR</i> mutations with varying tumour content. Thus it helps in triaging the cases judiciously before proceeding with multigene testing.</p>","PeriodicalId":14157,"journal":{"name":"International Journal of Experimental Pathology","volume":null,"pages":null},"PeriodicalIF":3.0,"publicationDate":"2024-05-08","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"140876421","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Túlio de Almeida Hermes, Paula Fratini, Beatriz Godinho Nascimento, Laís Leite Ferreira, Giuliana Petri, Fernando Luiz Affonso Fonseca, Alzira Alves de Siqueira Carvalho, David Feder
Duchenne muscular dystrophy (DMD) occurs due to genetic mutations that lead to a deficiency in dystrophin production and consequent progressive degeneration of skeletal muscle fibres, through oxidative stress and an exacerbated inflammatory process. The flavonoid trilobatin (TLB) demonstrates antioxidant and anti-inflammatory potential. Its high safety profile and effective action make it a potent therapy for the process of dystrophic muscle myonecrosis. Thus, we sought to investigate the action of TLB on damage in a DMD model, the mdx mouse. Eight-week-old male animals were treated with 160 mg/kg/day of trilobatin for 8 weeks. Control animals were treated with saline. Following treatment, muscle strength, serum creatine kinase (CK) levels, histopathology (necrotic myofibres, regenerated fibres/central nuclei, Feret's diameter and inflammatory area) and the levels of catalase and NF-κB (western blotting) of the quadriceps (QUA), diaphragm (DIA) and tibialis anterior (TA) muscles were measured. TLB was able to significantly increase muscle strength and reduce serum CK levels in dystrophic animals. The QUA of mdx mice showed a reduction in catalase and the number of fibres with a centralized nucleus after treatment with TLB. In the DIA of dystrophic animals, TLB reduced the necrotic myofibres, inflammatory area and NF-κB and increased the number of regenerated fibres and the total fibre diameter. In TA, TLB increased the number of regenerated fibres and reduced catalase levels in these animals. It is concluded that in the mdx experimental model, treatment with TLB was beneficial in the treatment of DMD.
{"title":"Trilobatin contributes to the improvement of myopathy in a mouse model of Duchenne muscular dystrophy","authors":"Túlio de Almeida Hermes, Paula Fratini, Beatriz Godinho Nascimento, Laís Leite Ferreira, Giuliana Petri, Fernando Luiz Affonso Fonseca, Alzira Alves de Siqueira Carvalho, David Feder","doi":"10.1111/iep.12502","DOIUrl":"10.1111/iep.12502","url":null,"abstract":"<p>Duchenne muscular dystrophy (DMD) occurs due to genetic mutations that lead to a deficiency in dystrophin production and consequent progressive degeneration of skeletal muscle fibres, through oxidative stress and an exacerbated inflammatory process. The flavonoid trilobatin (TLB) demonstrates antioxidant and anti-inflammatory potential. Its high safety profile and effective action make it a potent therapy for the process of dystrophic muscle myonecrosis. Thus, we sought to investigate the action of TLB on damage in a DMD model, the mdx mouse. Eight-week-old male animals were treated with 160 mg/kg/day of trilobatin for 8 weeks. Control animals were treated with saline. Following treatment, muscle strength, serum creatine kinase (CK) levels, histopathology (necrotic myofibres, regenerated fibres/central nuclei, Feret's diameter and inflammatory area) and the levels of catalase and NF-κB (western blotting) of the quadriceps (QUA), diaphragm (DIA) and tibialis anterior (TA) muscles were measured. TLB was able to significantly increase muscle strength and reduce serum CK levels in dystrophic animals. The QUA of mdx mice showed a reduction in catalase and the number of fibres with a centralized nucleus after treatment with TLB. In the DIA of dystrophic animals, TLB reduced the necrotic myofibres, inflammatory area and NF-κB and increased the number of regenerated fibres and the total fibre diameter. In TA, TLB increased the number of regenerated fibres and reduced catalase levels in these animals. It is concluded that in the mdx experimental model, treatment with TLB was beneficial in the treatment of DMD.</p>","PeriodicalId":14157,"journal":{"name":"International Journal of Experimental Pathology","volume":null,"pages":null},"PeriodicalIF":3.0,"publicationDate":"2024-03-13","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"140110222","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}