ENGINEERED NANOBODIES WITH PROGRAMMABLE TARGET ANTIGEN PROTEOLYSIS (PTAP) FUSIONS REGULATE INTRACELLULAR ALPHA-SYNUCLEIN IN VITRO AND IN VIVO.

Autor:	Chatterjee D; Department of Neurological Sciences, Rush University Medical Center, Chicago, IL 60612.; Department of Neurology, Feinberg School of Medicine, Northwestern University, Chicago, IL 60611., D'Brant LY; Regenerative Research Foundation, Neural Stem Cell Institute, Rensselaer, NY 12144., Hiller BM; Department of Neurological Sciences, Rush University Medical Center, Chicago, IL 60612., Marmion DJ; Department of Neurological Sciences, Rush University Medical Center, Chicago, IL 60612., Sandoval IM; Department of Translational Neuroscience, Barrow Neurological Institute, Phoenix, AZ 85013., Luk KC; Department of Pathology and Laboratory Medicine, University of Pennsylvania, Philadelphia, PA 19147., Manfredsson FP; Department of Translational Neuroscience, Barrow Neurological Institute, Phoenix, AZ 85013., Messer A; Regenerative Research Foundation, Neural Stem Cell Institute, Rensselaer, NY 12144.; Department of Biomedical Sciences, University at Albany, Albany, NY 12208., Kordower JH; Department of Neurological Sciences, Rush University Medical Center, Chicago, IL 60612.; ASU-Banner Neurodegenerative Disease Research Center and School of Life Sciences, Arizona State University, Tempe, AZ 85281., Butler DC; Regenerative Research Foundation, Neural Stem Cell Institute, Rensselaer, NY 12144.; Department of Biomedical Sciences, University at Albany, Albany, NY 12208.
Jazyk:	angličtina
Zdroj:	Research square [Res Sq] 2024 Mar 28. Date of Electronic Publication: 2024 Mar 28.
DOI:	10.21203/rs.3.rs-4088206/v1
Abstrakt:	Alpha-synuclein (αSyn) aggregation and the formation of Lewy pathology (LP) is a foundational pathophysiological phenomenon in synucleinopathies. Delivering therapeutic single-chain and single-domain antibodies that bind pathogenic targets can disrupt intracellular aggregation. The fusion of antibody fragments to a negatively-charged proteasomal targeting motif (PEST) creates bifunctional constructs that enhance both solubility and turnover. With sequence-specific point mutations of PEST sequences that modulate proteasomal degradation efficiency, we report the creation of Programmable Target Antigen Proteolysis (PTAP) technology that can provide graded control over the levels of target antigens. We have previously demonstrated our lead anti-αSyn intrabody, VH14-PEST, is capable of reducing the pathological burden of synucleinopathy in vitro and in vivo . Here, we report a family of fully humanized VH14-PTAP constructs for controllable, therapeutic targeting of intracellular α-Syn. In cells, we demonstrate successful target engagement and efficacy of VH14-hPEST intrabodies, and validate proof-of-principle in human cells using 3D human organoids derived from PD-patient induced pluripotent stem cells (iPSC). In two synuclein-based rat models, PTAP intrabodies attenuated nigral αSyn pathology, preserved nigrostriatal dopaminergic tone, and slowed the propagation of αSyn pathology. These data demonstrate the potency of intracellular αSyn targeting as a method to alleviate pathology and highlight the potential clinical utility of PTAP intrabodies. Competing Interests: Conflicts of Interest The authors A.M. and D.C.B are listed as inventors on a patent application (WO2022103977A1) filed by the Regenerative Research Foundation related to this work. J.H.K has received commercial support from Ryne Bio, Abbvie, Amydis, Inc., Biogen, Renovacor, Clintrex, Inhibikase Therapeutics, Inc., Mark Therapeutics, and Encora Therapeutics.
Databáze:	MEDLINE
Externí odkaz:	Zobrazit plný text záznamu