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Targeted and selective knockout of the TLQP-21 neuropeptide unmasks its unique role in energy homeostasis
OBJECTIVE: Pro-peptide precursors are processed into biologically active peptide hormones or neurotransmitters, each playing an essential role in physiology and disease. Genetic loss of function of a pro-peptide precursor results in the simultaneous ablation of all biologically-active peptides withi...
Autores principales: | , , , , , , , , , , , , , , , , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
Elsevier
2023
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10400922/ https://www.ncbi.nlm.nih.gov/pubmed/37482186 http://dx.doi.org/10.1016/j.molmet.2023.101781 |
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author | Sahu, Bhavani S. Razzoli, Maria McGonigle, Seth Pallais, Jean Pierre Nguyen, Megin E. Sadahiro, Masato Jiang, Cheng Lin, Wei-Jye Kelley, Kevin A. Rodriguez, Pedro Mansk, Rachel Cero, Cheryl Caviola, Giada Palanza, Paola Rao, Loredana Beetch, Megan Alejandro, Emilyn Sham, Yuk Y. Frontini, Andrea Salton, Stephen R. Bartolomucci, Alessandro |
author_facet | Sahu, Bhavani S. Razzoli, Maria McGonigle, Seth Pallais, Jean Pierre Nguyen, Megin E. Sadahiro, Masato Jiang, Cheng Lin, Wei-Jye Kelley, Kevin A. Rodriguez, Pedro Mansk, Rachel Cero, Cheryl Caviola, Giada Palanza, Paola Rao, Loredana Beetch, Megan Alejandro, Emilyn Sham, Yuk Y. Frontini, Andrea Salton, Stephen R. Bartolomucci, Alessandro |
author_sort | Sahu, Bhavani S. |
collection | PubMed |
description | OBJECTIVE: Pro-peptide precursors are processed into biologically active peptide hormones or neurotransmitters, each playing an essential role in physiology and disease. Genetic loss of function of a pro-peptide precursor results in the simultaneous ablation of all biologically-active peptides within that precursor, often leading to a composite phenotype that can be difficult to align with the loss of specific peptide components. Due to this biological constraint and technical limitations, mice carrying the selective ablation of individual peptides encoded by pro-peptide precursor genes, while leaving the other peptides unaffected, have remained largely unaddressed. METHODS: We developed and characterized a mouse model carrying the selective knockout of the TLQP-21 neuropeptide (ΔTLQP-21) encoded by the Vgf gene. To achieve this goal, we used a knowledge-based approach by mutating a codon in the Vgf sequence leading to the substitution of the C-terminal Arginine of TLQP-21, which is the pharmacophore as well as an essential cleavage site from its precursor, into Alanine (R(21)→A). RESULTS: We provide several independent validations of this mouse, including a novel in-gel digestion targeted mass spectrometry identification of the unnatural mutant sequence, exclusive to the mutant mouse. ΔTLQP-21 mice do not manifest gross behavioral and metabolic abnormalities and reproduce well, yet they have a unique metabolic phenotype characterized by an environmental temperature-dependent resistance to diet-induced obesity and activation of the brown adipose tissue. CONCLUSIONS: The ΔTLQP-21 mouse line can be a valuable resource to conduct mechanistic studies on the necessary role of TLQP-21 in physiology and disease, while also serving as a platform to test the specificity of novel antibodies or immunoassays directed at TLQP-21. Our approach also has far-reaching implications by informing the development of knowledge-based genetic engineering approaches to generate selective loss of function of other peptides encoded by pro-hormones genes, leaving all other peptides within the pro-protein precursor intact and unmodified. |
format | Online Article Text |
id | pubmed-10400922 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2023 |
publisher | Elsevier |
record_format | MEDLINE/PubMed |
spelling | pubmed-104009222023-08-05 Targeted and selective knockout of the TLQP-21 neuropeptide unmasks its unique role in energy homeostasis Sahu, Bhavani S. Razzoli, Maria McGonigle, Seth Pallais, Jean Pierre Nguyen, Megin E. Sadahiro, Masato Jiang, Cheng Lin, Wei-Jye Kelley, Kevin A. Rodriguez, Pedro Mansk, Rachel Cero, Cheryl Caviola, Giada Palanza, Paola Rao, Loredana Beetch, Megan Alejandro, Emilyn Sham, Yuk Y. Frontini, Andrea Salton, Stephen R. Bartolomucci, Alessandro Mol Metab Original Article OBJECTIVE: Pro-peptide precursors are processed into biologically active peptide hormones or neurotransmitters, each playing an essential role in physiology and disease. Genetic loss of function of a pro-peptide precursor results in the simultaneous ablation of all biologically-active peptides within that precursor, often leading to a composite phenotype that can be difficult to align with the loss of specific peptide components. Due to this biological constraint and technical limitations, mice carrying the selective ablation of individual peptides encoded by pro-peptide precursor genes, while leaving the other peptides unaffected, have remained largely unaddressed. METHODS: We developed and characterized a mouse model carrying the selective knockout of the TLQP-21 neuropeptide (ΔTLQP-21) encoded by the Vgf gene. To achieve this goal, we used a knowledge-based approach by mutating a codon in the Vgf sequence leading to the substitution of the C-terminal Arginine of TLQP-21, which is the pharmacophore as well as an essential cleavage site from its precursor, into Alanine (R(21)→A). RESULTS: We provide several independent validations of this mouse, including a novel in-gel digestion targeted mass spectrometry identification of the unnatural mutant sequence, exclusive to the mutant mouse. ΔTLQP-21 mice do not manifest gross behavioral and metabolic abnormalities and reproduce well, yet they have a unique metabolic phenotype characterized by an environmental temperature-dependent resistance to diet-induced obesity and activation of the brown adipose tissue. CONCLUSIONS: The ΔTLQP-21 mouse line can be a valuable resource to conduct mechanistic studies on the necessary role of TLQP-21 in physiology and disease, while also serving as a platform to test the specificity of novel antibodies or immunoassays directed at TLQP-21. Our approach also has far-reaching implications by informing the development of knowledge-based genetic engineering approaches to generate selective loss of function of other peptides encoded by pro-hormones genes, leaving all other peptides within the pro-protein precursor intact and unmodified. Elsevier 2023-07-21 /pmc/articles/PMC10400922/ /pubmed/37482186 http://dx.doi.org/10.1016/j.molmet.2023.101781 Text en © 2023 The Author(s) https://creativecommons.org/licenses/by-nc-nd/4.0/This is an open access article under the CC BY-NC-ND license (http://creativecommons.org/licenses/by-nc-nd/4.0/). |
spellingShingle | Original Article Sahu, Bhavani S. Razzoli, Maria McGonigle, Seth Pallais, Jean Pierre Nguyen, Megin E. Sadahiro, Masato Jiang, Cheng Lin, Wei-Jye Kelley, Kevin A. Rodriguez, Pedro Mansk, Rachel Cero, Cheryl Caviola, Giada Palanza, Paola Rao, Loredana Beetch, Megan Alejandro, Emilyn Sham, Yuk Y. Frontini, Andrea Salton, Stephen R. Bartolomucci, Alessandro Targeted and selective knockout of the TLQP-21 neuropeptide unmasks its unique role in energy homeostasis |
title | Targeted and selective knockout of the TLQP-21 neuropeptide unmasks its unique role in energy homeostasis |
title_full | Targeted and selective knockout of the TLQP-21 neuropeptide unmasks its unique role in energy homeostasis |
title_fullStr | Targeted and selective knockout of the TLQP-21 neuropeptide unmasks its unique role in energy homeostasis |
title_full_unstemmed | Targeted and selective knockout of the TLQP-21 neuropeptide unmasks its unique role in energy homeostasis |
title_short | Targeted and selective knockout of the TLQP-21 neuropeptide unmasks its unique role in energy homeostasis |
title_sort | targeted and selective knockout of the tlqp-21 neuropeptide unmasks its unique role in energy homeostasis |
topic | Original Article |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10400922/ https://www.ncbi.nlm.nih.gov/pubmed/37482186 http://dx.doi.org/10.1016/j.molmet.2023.101781 |
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