Targeted and selective knockout of the TLQP-21 neuropeptide unmasks its unique role in energy homeostasis.

Autor:	Sahu BS; Department of Integrative Biology and Physiology, University of Minnesota, Minneapolis, MN, 55455, USA., Razzoli M; Department of Integrative Biology and Physiology, University of Minnesota, Minneapolis, MN, 55455, USA., McGonigle S; Department of Integrative Biology and Physiology, University of Minnesota, Minneapolis, MN, 55455, USA., Pallais JP; Department of Integrative Biology and Physiology, University of Minnesota, Minneapolis, MN, 55455, USA., Nguyen ME; Department of Integrative Biology and Physiology, University of Minnesota, Minneapolis, MN, 55455, USA., Sadahiro M; Nash Family Department of Neuroscience, Icahn School of Medicine at Mount Sinai, New York, NY, 10029, USA., Jiang C; Nash Family Department of Neuroscience, Icahn School of Medicine at Mount Sinai, New York, NY, 10029, USA., Lin WJ; Nash Family Department of Neuroscience, Icahn School of Medicine at Mount Sinai, New York, NY, 10029, USA., Kelley KA; Department of Cell, Developmental and Regenerative Biology, Icahn School of Medicine at Mount Sinai, New York, NY, 10029, USA., Rodriguez P; Department of Integrative Biology and Physiology, University of Minnesota, Minneapolis, MN, 55455, USA., Mansk R; Department of Integrative Biology and Physiology, University of Minnesota, Minneapolis, MN, 55455, USA., Cero C; Department of Integrative Biology and Physiology, University of Minnesota, Minneapolis, MN, 55455, USA., Caviola G; Department of Medicine and Surgery, University of Parma, 43120, Parma, Italy., Palanza P; Department of Medicine and Surgery, University of Parma, 43120, Parma, Italy., Rao L; Department of Life and Environmental Sciences, Universita' Politecnica delle Marche, Ancona, 60131, Italy., Beetch M; Department of Integrative Biology and Physiology, University of Minnesota, Minneapolis, MN, 55455, USA., Alejandro E; Department of Integrative Biology and Physiology, University of Minnesota, Minneapolis, MN, 55455, USA., Sham YY; Department of Integrative Biology and Physiology, University of Minnesota, Minneapolis, MN, 55455, USA., Frontini A; Department of Life and Environmental Sciences, Universita' Politecnica delle Marche, Ancona, 60131, Italy., Salton SR; Department of Cell, Developmental and Regenerative Biology, Icahn School of Medicine at Mount Sinai, New York, NY, 10029, USA., Bartolomucci A; Department of Integrative Biology and Physiology, University of Minnesota, Minneapolis, MN, 55455, USA. Electronic address: abartolo@umn.edu.
Jazyk:	angličtina
Zdroj:	Molecular metabolism [Mol Metab] 2023 Oct; Vol. 76, pp. 101781. Date of Electronic Publication: 2023 Jul 21.
DOI:	10.1016/j.molmet.2023.101781
Abstrakt:	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. Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper. (Copyright © 2023 The Author(s). Published by Elsevier GmbH.. All rights reserved.)
Databáze:	MEDLINE
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