Cardiac hypertrophy is an adaptive cardiac response to heart stress. Sustained cardiac hypertrophy indicates higher risk of heart failure. Ca2+/calmodulin-dependent protein kinase II (CaMKII) has been proved to be a key regulator of cardiac hypertrophy, but its mechanism remains largely unknown. Our study proposed to explore the regulatory mechanism of CaMKII in cardiac hypertrophy. We validated that CaMKII was upregulated in cardiac hypertrophy models in vivo and in vitro, and that knockdown of CaMKII attenuated Ang II-induced cardiac hypertrophy in vitro. Furtherly, we demonstrated that STAT3 was highly expressed in cardiac hypertrophy and could stimulate the transactivation of CaMKII. Moreover, we predicted through Targetscan and confirmed that miR-625-5p targeted and inhibited STAT3 so as to reduce the expression of CaMKII. Interestingly, we also found that miR-625-5p directly targeted CaMKII and inhibited its expression. Rescue assays suggested that miR-625-5p attenuated Ang II-induced cardiac hypertrophy through CaMKII/STAT3. Consequently, this study elucidated that miR-625-5p inhibited cardiac hypertrophy through targeting STAT3 and CaMKII, suggesting miR-625-5p as a novel negative regulator of cardiac hypertrophy.
{"title":"MiR-625-5p inhibits cardiac hypertrophy through targeting STAT3 and CaMKII.","authors":"Kefeng Cai, Huiqin Chen","doi":"10.1089/humc.2019.087","DOIUrl":"https://doi.org/10.1089/humc.2019.087","url":null,"abstract":"Cardiac hypertrophy is an adaptive cardiac response to heart stress. Sustained cardiac hypertrophy indicates higher risk of heart failure. Ca2+/calmodulin-dependent protein kinase II (CaMKII) has been proved to be a key regulator of cardiac hypertrophy, but its mechanism remains largely unknown. Our study proposed to explore the regulatory mechanism of CaMKII in cardiac hypertrophy. We validated that CaMKII was upregulated in cardiac hypertrophy models in vivo and in vitro, and that knockdown of CaMKII attenuated Ang II-induced cardiac hypertrophy in vitro. Furtherly, we demonstrated that STAT3 was highly expressed in cardiac hypertrophy and could stimulate the transactivation of CaMKII. Moreover, we predicted through Targetscan and confirmed that miR-625-5p targeted and inhibited STAT3 so as to reduce the expression of CaMKII. Interestingly, we also found that miR-625-5p directly targeted CaMKII and inhibited its expression. Rescue assays suggested that miR-625-5p attenuated Ang II-induced cardiac hypertrophy through CaMKII/STAT3. Consequently, this study elucidated that miR-625-5p inhibited cardiac hypertrophy through targeting STAT3 and CaMKII, suggesting miR-625-5p as a novel negative regulator of cardiac hypertrophy.","PeriodicalId":51315,"journal":{"name":"Human Gene Therapy Clinical Development","volume":"29 1","pages":""},"PeriodicalIF":0.0,"publicationDate":"2019-12-16","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"74608911","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Background Hepatocellular carcinoma (HCC) is a most common malignancy in liver and is one of the leading causes of cancer-induced deaths over the world. Circular RNAs (circRNAs) have been proven to be related to cancer initiation and progression in mounting reports. However, research on the role of circRNAs in human cancers, including HCC, is still in its infancy. CircVAPA has been unmasked as oncogenic in colorectal cancer. Yet the function of circVAPA in HCC has never been elucidated. Materials and Methods CircVAPA, miR-377-3p and PSAP mRNA expression levels were detected by RT-qPCR. PSAP protein levels were measured by western blot. Cell proliferation was evaluated by CCK-8, colony formation and EdU assays. Binding capacity was assessed by dual-luciferase reporter assay. Results CircVAPA was upregulated in HCC cell lines and circVAPA depletion was associated with decreased HCC cell proliferation. CircVAPA promotes PSAP expression via sequestering miR-377-3p. The suppression of HCC cell proliferation caused by circVAPA silence was revived by PSAP overexpression. Conclusions This study revealed that circVAPA contributes to HCC cell proliferation via sponging miR-377-3p and thereby disinhibiting PSAP, shedding light on a new insight into HCC initiation and progression.
{"title":"Circular RNA circVAPA promotes cell proliferation in hepatocellular carcinoma.","authors":"Caifeng Liu, X. Zhong, Jun Li, Feng Xu","doi":"10.1089/humc.2019.079","DOIUrl":"https://doi.org/10.1089/humc.2019.079","url":null,"abstract":"Background Hepatocellular carcinoma (HCC) is a most common malignancy in liver and is one of the leading causes of cancer-induced deaths over the world. Circular RNAs (circRNAs) have been proven to be related to cancer initiation and progression in mounting reports. However, research on the role of circRNAs in human cancers, including HCC, is still in its infancy. CircVAPA has been unmasked as oncogenic in colorectal cancer. Yet the function of circVAPA in HCC has never been elucidated. Materials and Methods CircVAPA, miR-377-3p and PSAP mRNA expression levels were detected by RT-qPCR. PSAP protein levels were measured by western blot. Cell proliferation was evaluated by CCK-8, colony formation and EdU assays. Binding capacity was assessed by dual-luciferase reporter assay. Results CircVAPA was upregulated in HCC cell lines and circVAPA depletion was associated with decreased HCC cell proliferation. CircVAPA promotes PSAP expression via sequestering miR-377-3p. The suppression of HCC cell proliferation caused by circVAPA silence was revived by PSAP overexpression. Conclusions This study revealed that circVAPA contributes to HCC cell proliferation via sponging miR-377-3p and thereby disinhibiting PSAP, shedding light on a new insight into HCC initiation and progression.","PeriodicalId":51315,"journal":{"name":"Human Gene Therapy Clinical Development","volume":"1 1","pages":""},"PeriodicalIF":0.0,"publicationDate":"2019-12-16","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"88473439","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Ting He, Michelle S. Itano, L. F. Earley, N. E. Hall, N. Riddick, R. Samulski, Chengwen Li
Adeno-associated virus (AAV) vectors have become an important tool for delivering therapeutic genes for a wide range of neurological diseases. AAV serotypes possess differential cellular tropism in the central nervous system. Although several AAV serotypes or mutants have been reported to transduce the brain efficiently, conflicting data occurs across studies with the use of various rodent strains from different genetic backgrounds. Herein, we performed a systematic comparison of brain transduction properties among five AAV serotypes (AAV2, 5, 7, 8, and 9) in two common rodent strains (C57BL/6J and FVB/N), following local intra-striatal injection of AAV vectors encoding EGFP driven by the CBh promoter. Important differences were found regarding overall transduction efficiency, cellular tropism, and retrograde axonal transport among the AAV serotypes and between the mouse strains. We have further found loss of NeuN-immunoreactivity and microglia activation from AAV transduction in the different mouse strains. The important strain-specific differences from our study suggest that the genetic background of the mouse strains may affect AAV serotype transduction properties in the brain. This data can provide valuable information about how to choose an effective AAV vector for clinical application and interpret the data obtained from preclinical studies and clinical trials.
{"title":"The influence of murine genetic background in AAV transduction of the mouse brain.","authors":"Ting He, Michelle S. Itano, L. F. Earley, N. E. Hall, N. Riddick, R. Samulski, Chengwen Li","doi":"10.1089/humc.2019.030","DOIUrl":"https://doi.org/10.1089/humc.2019.030","url":null,"abstract":"Adeno-associated virus (AAV) vectors have become an important tool for delivering therapeutic genes for a wide range of neurological diseases. AAV serotypes possess differential cellular tropism in the central nervous system. Although several AAV serotypes or mutants have been reported to transduce the brain efficiently, conflicting data occurs across studies with the use of various rodent strains from different genetic backgrounds. Herein, we performed a systematic comparison of brain transduction properties among five AAV serotypes (AAV2, 5, 7, 8, and 9) in two common rodent strains (C57BL/6J and FVB/N), following local intra-striatal injection of AAV vectors encoding EGFP driven by the CBh promoter. Important differences were found regarding overall transduction efficiency, cellular tropism, and retrograde axonal transport among the AAV serotypes and between the mouse strains. We have further found loss of NeuN-immunoreactivity and microglia activation from AAV transduction in the different mouse strains. The important strain-specific differences from our study suggest that the genetic background of the mouse strains may affect AAV serotype transduction properties in the brain. This data can provide valuable information about how to choose an effective AAV vector for clinical application and interpret the data obtained from preclinical studies and clinical trials.","PeriodicalId":51315,"journal":{"name":"Human Gene Therapy Clinical Development","volume":"10 1","pages":""},"PeriodicalIF":0.0,"publicationDate":"2019-11-21","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"78993651","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
L. Tian, Zhiyong Su, Xiaobin Ma, Feng Wang, Yusong Guo
MiR-203 is known to target estrogen receptor α (ERα) in various cancer cell lines, such as MCF-7. However, whether miR-203 regulates ERα and contributes to the onset and progression of osteoarthritis (OA) is poorly understood. A combined protocol of the bilateral ovariectomy and the intra-articular monosodium iodoacetate (MIA) injection was applied to establish a postmenopausal OA model in rats. Real-time quantitative PCR was used to detect miR-203 and mRNAs and Western blotting was exploited to quantify the expressions on the protein level. ELISA assays were deployed to detect the expression of MMP-1, MMP-3, PGE2 and CTX-II in serum samples. Dual-luciferase reporter assay was utilized to confirm the direct binding of miR-203 on ERα in postmenopausal OA rats. Expression of miR-203 was elevated; while ERα mRNA and protein were down-regulated in postmenopausal OA rats, compared to sham rats. Dual-luciferase reporter assay confirmed miR-203 bound and negatively regulated ERα, resulting in promoted cellular inflammation and cartilage destruction in postmenopausal OA rats. Suppression of miR-203 using a specific inhibitor ameliorated cartilage degradation in postmenopausal OA rats. MiR-203 is pivotal in the onset and progression of OA in the postmenopausal rat model, and holds promise for a therapeutic target of OA treatment.
{"title":"Inhibition of miR-203 ameliorates osteoarthritis cartilage degradation in the postmenopausal rat model: involvement of estrogen receptor α.","authors":"L. Tian, Zhiyong Su, Xiaobin Ma, Feng Wang, Yusong Guo","doi":"10.1089/humc.2019.101","DOIUrl":"https://doi.org/10.1089/humc.2019.101","url":null,"abstract":"MiR-203 is known to target estrogen receptor α (ERα) in various cancer cell lines, such as MCF-7. However, whether miR-203 regulates ERα and contributes to the onset and progression of osteoarthritis (OA) is poorly understood. A combined protocol of the bilateral ovariectomy and the intra-articular monosodium iodoacetate (MIA) injection was applied to establish a postmenopausal OA model in rats. Real-time quantitative PCR was used to detect miR-203 and mRNAs and Western blotting was exploited to quantify the expressions on the protein level. ELISA assays were deployed to detect the expression of MMP-1, MMP-3, PGE2 and CTX-II in serum samples. Dual-luciferase reporter assay was utilized to confirm the direct binding of miR-203 on ERα in postmenopausal OA rats. Expression of miR-203 was elevated; while ERα mRNA and protein were down-regulated in postmenopausal OA rats, compared to sham rats. Dual-luciferase reporter assay confirmed miR-203 bound and negatively regulated ERα, resulting in promoted cellular inflammation and cartilage destruction in postmenopausal OA rats. Suppression of miR-203 using a specific inhibitor ameliorated cartilage degradation in postmenopausal OA rats. MiR-203 is pivotal in the onset and progression of OA in the postmenopausal rat model, and holds promise for a therapeutic target of OA treatment.","PeriodicalId":51315,"journal":{"name":"Human Gene Therapy Clinical Development","volume":"34 1","pages":""},"PeriodicalIF":0.0,"publicationDate":"2019-11-11","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"81732752","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2019-09-17DOI: 10.1089/humc.2019.29047.int
Kevin Davies
{"title":"Looking into a Crystal Ball: An Interview with Ron Crystal.","authors":"Kevin Davies","doi":"10.1089/humc.2019.29047.int","DOIUrl":"https://doi.org/10.1089/humc.2019.29047.int","url":null,"abstract":"","PeriodicalId":51315,"journal":{"name":"Human Gene Therapy Clinical Development","volume":"27 1","pages":"97-101"},"PeriodicalIF":0.0,"publicationDate":"2019-09-17","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"88055506","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
The purpose of this study was to investigate the expression level of microRNA-4513 (miR-4513) in gastric cancer, and the underlying mechanisms of miR-4513 in regulating gastric cancer (GC) progression. Quantitative real time-polymerase chain reaction (qRT-PCR) was performed to measure the expression level of miR-4513 in GC cells. Transfection efficacy of synthetic miRNAs was examined by qRT-PCR. After synthetic miRNAs transfection, cell counting kit-8 assay and transwell invasion assay was conducted to measure biological changes in these groups. Key molecular expression level involved in epithelial-mesenchymal transition (EMT) was analyzed by western blot. Bioinformatic analysis and western blot were performed to investigate the connection between miR-4513 and lysine acetyltransferase 6B (KAT6B). qRT-PCR results showed miR-4513 expression level was upregulated in GC cell lines. Downregulate miR-4513 expression inhibited GC cell proliferation, invasion, and EMT. KAT6B was validated as a direct target of miR-4513. In addition, KAT6B expression level can be upregulated by miR-4513 inhibitor. Collectively, we showed miR-4513 involved in regulating the biological function of GC cells via KAT6B. In addition, miR-4513 may serve as a potential target for the molecular therapy of GC.
{"title":"miR-4513 promotes gastric cancer cell proliferation and epithelial-mesenchymal transition through targeting KAT6B.","authors":"Huiming Ding, Yuhua Shi, Xiaobing Liu, A. Qiu","doi":"10.1089/humc.2019.094","DOIUrl":"https://doi.org/10.1089/humc.2019.094","url":null,"abstract":"The purpose of this study was to investigate the expression level of microRNA-4513 (miR-4513) in gastric cancer, and the underlying mechanisms of miR-4513 in regulating gastric cancer (GC) progression. Quantitative real time-polymerase chain reaction (qRT-PCR) was performed to measure the expression level of miR-4513 in GC cells. Transfection efficacy of synthetic miRNAs was examined by qRT-PCR. After synthetic miRNAs transfection, cell counting kit-8 assay and transwell invasion assay was conducted to measure biological changes in these groups. Key molecular expression level involved in epithelial-mesenchymal transition (EMT) was analyzed by western blot. Bioinformatic analysis and western blot were performed to investigate the connection between miR-4513 and lysine acetyltransferase 6B (KAT6B). qRT-PCR results showed miR-4513 expression level was upregulated in GC cell lines. Downregulate miR-4513 expression inhibited GC cell proliferation, invasion, and EMT. KAT6B was validated as a direct target of miR-4513. In addition, KAT6B expression level can be upregulated by miR-4513 inhibitor. Collectively, we showed miR-4513 involved in regulating the biological function of GC cells via KAT6B. In addition, miR-4513 may serve as a potential target for the molecular therapy of GC.","PeriodicalId":51315,"journal":{"name":"Human Gene Therapy Clinical Development","volume":"30 1","pages":""},"PeriodicalIF":0.0,"publicationDate":"2019-09-16","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"81321708","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2019-09-01Epub Date: 2019-06-21DOI: 10.1089/humc.2019.042
Houria Bachtarzi, Tim Farries
In vivo viral gene therapy and somatic cell therapy products (whether autologous, allogeneic, or xenogeneic) that have been subjected to an ex vivo gene transfer procedure will normally be classified as genetically modified organisms (GMOs) in Europe, not just the gene transfer vectors used in their construction. These products are, therefore, expected to fulfill certain environmental requirements with regard to the biosafety aspects of their clinical use (which may be subject to review by government departments responsible for environmental affairs). In the European Union (EU), clinical trials using GMOs generally require three levels of review (in addition to local review processes), which are often performed by separate national agencies. In this study, the principles under which certain EU member states control use of the GMOs in clinical trials under the definitions of either "contained use" or "deliberate release" will be discussed and evaluated from a scientific and a regulatory perspective, with comparisons with non-EU expectations as described by the U.S. Food and Drug Administration and the Japanese living modified organisms (LMOs) regulations. For the latter, an understanding of the criteria under which LMOs exemptions apply, notably with respect to the nature of the viral construct used, the manufacturing process, and demonstration that there is no detectable residual replication-competent virus in the final gene-modified cells, is of paramount importance. Building on the existing European, U.S., and Japanese experience with GMOs/LMOs within the context of experimental gene and cell therapies, a through reflection on, and harmonization of, the current global GMO framework is needed to avoid unnecessary delays in clinical development and to ensure a smooth and a rapid access by patients to innovative life-saving therapies.
{"title":"The Genetically Modified Organism Medicinal Framework in Europe, United States, and Japan: Underlying Scientific Principles and Considerations Toward the Development of Gene Therapy and Genetically Modified Cell-Based Products.","authors":"Houria Bachtarzi, Tim Farries","doi":"10.1089/humc.2019.042","DOIUrl":"https://doi.org/10.1089/humc.2019.042","url":null,"abstract":"<p><p><i>In vivo</i> viral gene therapy and somatic cell therapy products (whether autologous, allogeneic, or xenogeneic) that have been subjected to an <i>ex vivo</i> gene transfer procedure will normally be classified as genetically modified organisms (GMOs) in Europe, not just the gene transfer vectors used in their construction. These products are, therefore, expected to fulfill certain environmental requirements with regard to the biosafety aspects of their clinical use (which may be subject to review by government departments responsible for environmental affairs). In the European Union (EU), clinical trials using GMOs generally require three levels of review (in addition to local review processes), which are often performed by separate national agencies. In this study, the principles under which certain EU member states control use of the GMOs in clinical trials under the definitions of either \"contained use\" or \"deliberate release\" will be discussed and evaluated from a scientific and a regulatory perspective, with comparisons with non-EU expectations as described by the U.S. Food and Drug Administration and the Japanese living modified organisms (LMOs) regulations. For the latter, an understanding of the criteria under which LMOs exemptions apply, notably with respect to the nature of the viral construct used, the manufacturing process, and demonstration that there is no detectable residual replication-competent virus in the final gene-modified cells, is of paramount importance. Building on the existing European, U.S., and Japanese experience with GMOs/LMOs within the context of experimental gene and cell therapies, a through reflection on, and harmonization of, the current global GMO framework is needed to avoid unnecessary delays in clinical development and to ensure a smooth and a rapid access by patients to innovative life-saving therapies.</p>","PeriodicalId":51315,"journal":{"name":"Human Gene Therapy Clinical Development","volume":"30 3","pages":"114-128"},"PeriodicalIF":0.0,"publicationDate":"2019-09-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1089/humc.2019.042","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"37262836","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2019-09-01DOI: 10.1089/humc.2019.29049.int
J. Mendell, James M. Wilson
{"title":"Breakthrough to Bedside: Bringing Gene Therapy to Neuromuscular Diseases.","authors":"J. Mendell, James M. Wilson","doi":"10.1089/humc.2019.29049.int","DOIUrl":"https://doi.org/10.1089/humc.2019.29049.int","url":null,"abstract":"","PeriodicalId":51315,"journal":{"name":"Human Gene Therapy Clinical Development","volume":"47 1","pages":""},"PeriodicalIF":0.0,"publicationDate":"2019-09-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"76640429","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
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