Genetic Deletion of DNAJB3 Using CRISPR-Cas9, Produced Discordant Phenotypes
<p dir="ltr">Several pathways and/or genes have been shown to be dysregulated in obesity-induced insulin resistance (IR) and type 2 diabetes (T2D). We previously showed, for the first time, impaired expression of DNAJB3 mRNA and protein in subjects with obesity, which was concomitant...
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2023
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| _version_ | 1864513507623436288 |
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| author | Shadi Nejat (19457212) |
| author2 | Kalhara R. Menikdiwela (18566698) Aliyah Efotte (19457215) Shane Scoggin (18566701) Bolormaa Vandanmagsar (4330651) Paul J. Thornalley (291723) Mohammed Dehbi (309033) Naima Moustaid-Moussa (66960) |
| author2_role | author author author author author author author |
| author_facet | Shadi Nejat (19457212) Kalhara R. Menikdiwela (18566698) Aliyah Efotte (19457215) Shane Scoggin (18566701) Bolormaa Vandanmagsar (4330651) Paul J. Thornalley (291723) Mohammed Dehbi (309033) Naima Moustaid-Moussa (66960) |
| author_role | author |
| dc.creator.none.fl_str_mv | Shadi Nejat (19457212) Kalhara R. Menikdiwela (18566698) Aliyah Efotte (19457215) Shane Scoggin (18566701) Bolormaa Vandanmagsar (4330651) Paul J. Thornalley (291723) Mohammed Dehbi (309033) Naima Moustaid-Moussa (66960) |
| dc.date.none.fl_str_mv | 2023-09-24T09:00:00Z |
| dc.identifier.none.fl_str_mv | 10.3390/genes14101857 |
| dc.relation.none.fl_str_mv | https://figshare.com/articles/journal_contribution/Genetic_Deletion_of_DNAJB3_Using_CRISPR-Cas9_Produced_Discordant_Phenotypes/26796082 |
| dc.rights.none.fl_str_mv | CC BY 4.0 info:eu-repo/semantics/openAccess |
| dc.subject.none.fl_str_mv | Biological sciences Genetics Biomedical and clinical sciences Clinical sciences Medical biochemistry and metabolomics obesity type 2 diabetes heat shock proteins DNAJB3 adipose tissue CRISPR-Cas9 inflammation insulin resistance glucose homeostasis ER stress |
| dc.title.none.fl_str_mv | Genetic Deletion of DNAJB3 Using CRISPR-Cas9, Produced Discordant Phenotypes |
| dc.type.none.fl_str_mv | Text Journal contribution info:eu-repo/semantics/publishedVersion text contribution to journal |
| description | <p dir="ltr">Several pathways and/or genes have been shown to be dysregulated in obesity-induced insulin resistance (IR) and type 2 diabetes (T2D). We previously showed, for the first time, impaired expression of DNAJB3 mRNA and protein in subjects with obesity, which was concomitant with increased metabolic stress. Restoring the normal expression of DNAJB3 attenuated metabolic stress and improved insulin signaling both in vivo and in vitro, suggesting a protective role of DNAJB3 against obesity and T2D. The precise underlying mechanisms remained, however, unclear. This study was designed to confirm the human studies in a mouse model of dietary obesity-induced insulin resistance, and, if validated, to understand the underlying mechanisms. We hypothesized that mice lacking DNAJB3 would be more prone to high-fat (HF)-diet-induced increase in body weight and body fat, inflammation, glucose intolerance and insulin resistance as compared with wild-type (WT) littermates. Three DNAJB3 knockout (KO) lines were generated (KO 30, 44 and 47), using CRISPR-Cas9. Male and female KO and WT mice were fed a HF diet (45% kcal fat) for 16 weeks. Body weight was measured biweekly, and a glucose tolerance test (GTT) and insulin tolerance test (ITT) were conducted at week 13 and 14, respectively. Body composition was determined monthly by nuclear magnetic resonance (NMR). Following euthanasia, white adipose tissue (WAT) and skeletal muscle were harvested for further analyses. Compared with WT mice, male and female KO 47 mice demonstrated higher body weight and fat mass. Similarly, KO 47 mice also showed a slower rate of glucose clearance in GTT that was consistent with decreased mRNA expression of the GLUT4 gene in WAT but not in the muscle. Both male and female KO 47 mice exhibited higher mRNA levels of the pro-inflammatory marker TNF-a in WAT only, whereas increased mRNA levels of MCP1 chemokine and the ER stress marker BiP/Grp78 were observed in male but not in female KO 47 mice. However, we did not observe the same changes in the other KO lines. Taken together, the phenotype of the DNAJB3 KO 47 mice was consistent with the metabolic changes and low levels of DNAJB3 reported in human subjects. These findings suggest that DNAJB3 may play an important role in metabolic functions and glucose homeostasis, which warrants further phenotyping and intervention studies in other KO 47 and other KO mice, as well as investigating this protein as a potential therapeutic target for obesity and T2D.</p><h2>Other Information</h2><p dir="ltr">Published in: Genes<br>License: <a href="https://creativecommons.org/licenses/by/4.0/" target="_blank">https://creativecommons.org/licenses/by/4.0/</a><br>See article on publisher's website: <a href="https://dx.doi.org/10.3390/genes14101857" target="_blank">https://dx.doi.org/10.3390/genes14101857</a></p> |
| eu_rights_str_mv | openAccess |
| id | Manara2_7af452a4b23728cdf3f90ed2fd302416 |
| identifier_str_mv | 10.3390/genes14101857 |
| network_acronym_str | Manara2 |
| network_name_str | Manara2 |
| oai_identifier_str | oai:figshare.com:article/26796082 |
| publishDate | 2023 |
| repository.mail.fl_str_mv | |
| repository.name.fl_str_mv | |
| repository_id_str | |
| rights_invalid_str_mv | CC BY 4.0 |
| spelling | Genetic Deletion of DNAJB3 Using CRISPR-Cas9, Produced Discordant PhenotypesShadi Nejat (19457212)Kalhara R. Menikdiwela (18566698)Aliyah Efotte (19457215)Shane Scoggin (18566701)Bolormaa Vandanmagsar (4330651)Paul J. Thornalley (291723)Mohammed Dehbi (309033)Naima Moustaid-Moussa (66960)Biological sciencesGeneticsBiomedical and clinical sciencesClinical sciencesMedical biochemistry and metabolomicsobesitytype 2 diabetesheat shock proteinsDNAJB3adipose tissueCRISPR-Cas9inflammationinsulin resistanceglucose homeostasisER stress<p dir="ltr">Several pathways and/or genes have been shown to be dysregulated in obesity-induced insulin resistance (IR) and type 2 diabetes (T2D). We previously showed, for the first time, impaired expression of DNAJB3 mRNA and protein in subjects with obesity, which was concomitant with increased metabolic stress. Restoring the normal expression of DNAJB3 attenuated metabolic stress and improved insulin signaling both in vivo and in vitro, suggesting a protective role of DNAJB3 against obesity and T2D. The precise underlying mechanisms remained, however, unclear. This study was designed to confirm the human studies in a mouse model of dietary obesity-induced insulin resistance, and, if validated, to understand the underlying mechanisms. We hypothesized that mice lacking DNAJB3 would be more prone to high-fat (HF)-diet-induced increase in body weight and body fat, inflammation, glucose intolerance and insulin resistance as compared with wild-type (WT) littermates. Three DNAJB3 knockout (KO) lines were generated (KO 30, 44 and 47), using CRISPR-Cas9. Male and female KO and WT mice were fed a HF diet (45% kcal fat) for 16 weeks. Body weight was measured biweekly, and a glucose tolerance test (GTT) and insulin tolerance test (ITT) were conducted at week 13 and 14, respectively. Body composition was determined monthly by nuclear magnetic resonance (NMR). Following euthanasia, white adipose tissue (WAT) and skeletal muscle were harvested for further analyses. Compared with WT mice, male and female KO 47 mice demonstrated higher body weight and fat mass. Similarly, KO 47 mice also showed a slower rate of glucose clearance in GTT that was consistent with decreased mRNA expression of the GLUT4 gene in WAT but not in the muscle. Both male and female KO 47 mice exhibited higher mRNA levels of the pro-inflammatory marker TNF-a in WAT only, whereas increased mRNA levels of MCP1 chemokine and the ER stress marker BiP/Grp78 were observed in male but not in female KO 47 mice. However, we did not observe the same changes in the other KO lines. Taken together, the phenotype of the DNAJB3 KO 47 mice was consistent with the metabolic changes and low levels of DNAJB3 reported in human subjects. These findings suggest that DNAJB3 may play an important role in metabolic functions and glucose homeostasis, which warrants further phenotyping and intervention studies in other KO 47 and other KO mice, as well as investigating this protein as a potential therapeutic target for obesity and T2D.</p><h2>Other Information</h2><p dir="ltr">Published in: Genes<br>License: <a href="https://creativecommons.org/licenses/by/4.0/" target="_blank">https://creativecommons.org/licenses/by/4.0/</a><br>See article on publisher's website: <a href="https://dx.doi.org/10.3390/genes14101857" target="_blank">https://dx.doi.org/10.3390/genes14101857</a></p>2023-09-24T09:00:00ZTextJournal contributioninfo:eu-repo/semantics/publishedVersiontextcontribution to journal10.3390/genes14101857https://figshare.com/articles/journal_contribution/Genetic_Deletion_of_DNAJB3_Using_CRISPR-Cas9_Produced_Discordant_Phenotypes/26796082CC BY 4.0info:eu-repo/semantics/openAccessoai:figshare.com:article/267960822023-09-24T09:00:00Z |
| spellingShingle | Genetic Deletion of DNAJB3 Using CRISPR-Cas9, Produced Discordant Phenotypes Shadi Nejat (19457212) Biological sciences Genetics Biomedical and clinical sciences Clinical sciences Medical biochemistry and metabolomics obesity type 2 diabetes heat shock proteins DNAJB3 adipose tissue CRISPR-Cas9 inflammation insulin resistance glucose homeostasis ER stress |
| status_str | publishedVersion |
| title | Genetic Deletion of DNAJB3 Using CRISPR-Cas9, Produced Discordant Phenotypes |
| title_full | Genetic Deletion of DNAJB3 Using CRISPR-Cas9, Produced Discordant Phenotypes |
| title_fullStr | Genetic Deletion of DNAJB3 Using CRISPR-Cas9, Produced Discordant Phenotypes |
| title_full_unstemmed | Genetic Deletion of DNAJB3 Using CRISPR-Cas9, Produced Discordant Phenotypes |
| title_short | Genetic Deletion of DNAJB3 Using CRISPR-Cas9, Produced Discordant Phenotypes |
| title_sort | Genetic Deletion of DNAJB3 Using CRISPR-Cas9, Produced Discordant Phenotypes |
| topic | Biological sciences Genetics Biomedical and clinical sciences Clinical sciences Medical biochemistry and metabolomics obesity type 2 diabetes heat shock proteins DNAJB3 adipose tissue CRISPR-Cas9 inflammation insulin resistance glucose homeostasis ER stress |