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US-12616735-B2 - Compositions and methods of controlling expression of thermogenin (UCP-1) in skeletal muscles

US12616735B2US 12616735 B2US12616735 B2US 12616735B2US-12616735-B2

Abstract

Described herein are pharmaceutical compositions containing genes encoding a positive regulatory domain zinc finger protein 16, a peroxisome proliferator-activated receptor gamma coactivator 1-alpha protein, and a bone morphogenetic protein 7. Also disclosed are methods of treating or managing obesity and diabetes by delivering these pharmaceutical compositions using ultrasound-targeted microbubble destruction.

Inventors

  • Raul Bastarrachea

Assignees

  • Raul Bastarrachea

Dates

Publication Date
20260505
Application Date
20190703

Claims (8)

  1. 1 . A pharmaceutical composition, comprising three expression cassettes capable of inducing UCP-1 expression, wherein: the composition is formulated for delivery to a skeletal muscle cell in a mammal; a first non-viral nucleic acid construct with a first expression cassette encodes a positive regulatory domain zinc finger protein 16; a second non-viral nucleic acid construct with a second expression cassette encodes a peroxisome proliferator-activated receptor gamma coactivator 1-alpha protein; and a third non-viral nucleic acid construct with a third expression cassette encodes a bone morphogenetic protein 7.
  2. 2 . The pharmaceutical composition of claim 1 , wherein the pharmaceutical composition is a microbubble suspension formed by mixing a plurality of lipids with a mixture of the first non-viral nucleic acid construct, the second nucleic acid construct, and the third nucleic acid construct.
  3. 3 . The pharmaceutical composition of claim 1 , wherein the pharmaceutical composition is a microbubble suspension of lipid-coated microbubbles containing a gas suitable for ultrasound-targeted microbubble destruction and a mixture of the first non-viral nucleic acid construct, the second nucleic acid construct, and the third nucleic acid construct.
  4. 4 . The pharmaceutical composition of claim 3 , wherein the gas is perfluoropropane.
  5. 5 . The pharmaceutical composition of claim 2 , wherein the plurality of lipids is cationic lipid.
  6. 6 . The pharmaceutical composition of claim 2 , wherein the plurality of lipids includes 1,2-dipalmitoyl-sn-glycero-3-phosphatidylcholine and 1,2-dipalmitoyl-sn-glycero-3-phosphatidyl ethanolamine.
  7. 7 . A pharmaceutical composition, comprising a non-viral expression vector containing three expression cassettes capable of inducing UCP-1 expression, wherein: the composition is formulated for delivery to a skeletal muscle cell in a mammal; a first expression cassette encoding a positive regulatory domain zinc finger protein 16; a second expression cassette encoding a peroxisome proliferator-activated receptor gamma coactivator 1-alpha protein; and a third expression cassette encoding a bone morphogenetic protein 7.
  8. 8 . The pharmaceutical composition of claim 7 , wherein the non-viral expression vector is a hyperactive piggyBac transposon-based vector.

Description

RELATED APPLICATION This application is claiming priority to and the benefit of U.S. Provisional Application Ser. No. 62/693,775 filed on Jul. 3, 2018, which is incorporated by reference in its entirety. TECHNICAL FIELD This disclosure relates to compositions and methods for managing obesity by controlling expression of mitochondrial uncoupling proteins in skeletal muscles of a mammalian subject. BACKGROUND Obesity is a common, complex, highly prevalent disorder currently affecting more than a third of the world's population. Obesity is closely associated with multiple metabolic disturbances including cardiometabolic diseases. Diet, exercise and behavior modifications remain the current cornerstones of obesity treatment and prevention, even though they work very poorly for successful long-term weight loss. Therefore, there is a desperate need of powerful strategies for the treatment of obesity to achieve long-term weight loss and maintenance. Brown adipose tissue is the main organ of adaptive nonshivering thermogenesis in humans. Brown adipocytes have a high mitochondrial content that contains a specialized protein—mitochondrial uncoupling protein (mUCP-1). This specialized protein uncouples ATP production from mitochondrial respiration and converts energy into heat. Gene therapy utilize both viral vectors and non-viral gene therapy. Non-viral gene delivery mechanisms are advantageous for certain diseases without potential complications of toxicity, altered immune response, and decreased capability to target specific cells. The focus of obesity-related gene therapy is to favor energy expenditure and lipolysis by modulating expression of appropriate gene targets to restore and maintain energy homeostasis. Ectopic expression of UCP-1 in skeletal muscles of transgenic mice has been shown to result in a phenotype characterized by increased energy expenditure, reduced body weight, reduced fat mass, improved glucose tolerance, decreased muscle energy efficiency, and altered substrate oxidation, as well as increased longevity. However, the usefulness of transgenic mice in determining the precise functional roles of cloned genes is limited by several technical factors such as variability in the copy number of transgenes inserted and potential functional alterations of neighboring cells. Random integration of transgenes within the genome is particularly worrisome. In theory, once integrated into the murine genome of transgenic mice, the injected DNA can manifest its function. However, as the insertion occurs at random, positional variegation or cell mutation effects may be considered, and both the function of endogenous genes might be affected by the insertion of a transgene as well as the expression of the transgene itself may also be severely compromised by surrounding elements. Delivery of adenoviruses containing cDNA of UCP-1 to the epididymal fat pads in mice induced localized fat depletion, improved glucose tolerance and decreased food intake in obese diabetic mice. However, there are disadvantages with the utilization of adenovirus vectors. As adenoviruses are non-integrating viruses, the transgene expression typically lasts about one to two months in non-dividing cells, and the expression time frame is much shorter in dividing cells. Therefore, transfer and expression are transient. Pre-existing immunity against adenoviruses in individuals may result in low levels of transgene delivery and expression. Viral vectors are immunogenic because the virus capsid and remaining viral proteins cause inflammation. In certain instances, adenovirus vectors cause patient death through their systemic delivery by triggering a massive inflammatory response that leads to disseminated intravascular coagulation and multi-organ failure. New strategies for weight loss treatment and prevention of obesity-related comorbidities are needed to successfully reverse or prevent the health complications caused by fat accumulation. SUMMARY Disclosed herein are compositions and methods addressing the shortcomings of the art, and may provide any number of additional or alternative advantages, including controlled expression of mitochondrial uncoupling proteins in skeletal muscles of a mammalian subject. Disclosed here are pharmaceutical compositions containing nucleic acid constructs coding PRDM16 (Positive Regulatory Domain Zinc Finger Protein 16), PPARGC-1α (Peroxisome proliferator-activated receptor gamma coactivator 1-alpha), and BMP7 (Bone morphogenetic protein 7). Also disclosed here are methods of delivery of the nucleic acid compositions containing expression cassettes coding PRDM16, PPARGC-1α, and BMP7 to skeletal muscle of a subject using a plurality of non-viral vectors and ultrasound-targeted microbubble destruction (UTMD) techniques. Also disclosed here are methods of delivery of the nucleic acid compositions containing expression cassettes coding PRDM16, PPARGC-1α, and BMP7 to skeletal muscle of a subject for the treatment of o