Programmable protein degraders enable selective knockdown of pathogenic β-catenin subpopulations in vitro and in vivo .

Autor:	Ye T; Robert F. Smith School of Chemical and Biomolecular Engineering, Cornell University, Ithaca, NY 14853 USA., Alamgir A; Robert F. Smith School of Chemical and Biomolecular Engineering, Cornell University, Ithaca, NY 14853 USA., Robertus CM; Nancy E. and Peter C. Meinig School of Biomedical Engineering, Cornell University, Ithaca, New York 14853 USA., Colina D; Biochemistry, Molecular and Cell Biology, Cornell University, Ithaca, NY 14853 USA., Monticello C; Nancy E. and Peter C. Meinig School of Biomedical Engineering, Cornell University, Ithaca, New York 14853 USA., Donahue TC; Robert F. Smith School of Chemical and Biomolecular Engineering, Cornell University, Ithaca, NY 14853 USA., Hong L; Department of Biomedical Engineering, Duke University, Durham, NC 27708 USA., Vincoff S; Department of Biomedical Engineering, Duke University, Durham, NC 27708 USA., Goel S; Department of Biomedical Engineering, Duke University, Durham, NC 27708 USA., Fekkes P; UbiquiTx, 750 Main Street, Cambridge, MA 02139 USA., Camargo LM; UbiquiTx, 750 Main Street, Cambridge, MA 02139 USA., Lam K; Genevant Sciences Corporation, 887 Great Northern Way, Vancouver, BC, V5T 4T5 Canada., Heyes J; Genevant Sciences Corporation, 887 Great Northern Way, Vancouver, BC, V5T 4T5 Canada., Putnam D; Robert F. Smith School of Chemical and Biomolecular Engineering, Cornell University, Ithaca, NY 14853 USA.; Nancy E. and Peter C. Meinig School of Biomedical Engineering, Cornell University, Ithaca, New York 14853 USA., Alabi CA; Robert F. Smith School of Chemical and Biomolecular Engineering, Cornell University, Ithaca, NY 14853 USA.; Nancy E. and Peter C. Meinig School of Biomedical Engineering, Cornell University, Ithaca, New York 14853 USA., Chatterjee P; Department of Biomedical Engineering, Duke University, Durham, NC 27708 USA.; Department of Computer Science, Duke University, Durham, NC 27708 USA.; Department of Biostatistics and Bioinformatics, Duke University, Durham, NC 27708 USA., DeLisa MP; Robert F. Smith School of Chemical and Biomolecular Engineering, Cornell University, Ithaca, NY 14853 USA.; Nancy E. and Peter C. Meinig School of Biomedical Engineering, Cornell University, Ithaca, New York 14853 USA.; Biochemistry, Molecular and Cell Biology, Cornell University, Ithaca, NY 14853 USA.; Cornell Institute of Biotechnology, Cornell University, Ithaca, NY 14853 USA.
Jazyk:	angličtina
Zdroj:	BioRxiv : the preprint server for biology [bioRxiv] 2024 Nov 11. Date of Electronic Publication: 2024 Nov 11.
DOI:	10.1101/2024.11.10.622803
Abstrakt:	Aberrant activation of Wnt signaling results in unregulated accumulation of cytosolic β-catenin, which subsequently enters the nucleus and promotes transcription of genes that contribute to cellular proliferation and malignancy. Here, we sought to eliminate pathogenic β-catenin from the cytosol using designer ubiquibodies (uAbs), chimeric proteins composed of an E3 ubiquitin ligase and a target-binding domain that redirect intracellular proteins to the proteasome for degradation. To accelerate uAb development, we leveraged a protein language model (pLM)-driven algorithm called SaLT&PepPr to computationally design "guide" peptides with affinity for β-catenin, which were subsequently fused to the catalytic domain of a human E3 called C-terminus of Hsp70-interacting protein (CHIP). Expression of the resulting peptide-guided uAbs in colorectal cancer cells led to the identification of several designs that significantly reduced the abnormally stable pool of free β-catenin in the cytosol and nucleus while preserving the normal membrane-associated subpopulation. This selective knockdown of pathogenic β-catenin suppressed Wnt/β-catenin signaling and impaired tumor cell survival and proliferation. Furthermore, one of the best degraders selectively decreased cytosolic but not membrane-associated β-catenin levels in livers of BALB/c mice following delivery as a lipid nanoparticle (LNP)-encapsulated mRNA. Collectively, these findings reveal the unique ability of uAbs to selectively eradicate abnormal proteins in vitro and in vivo and open the door to peptide-programmable biologic modulators of other disease-causing proteins. Competing Interests: Competing Interests Statement. M.P.D. and P.C. have financial interests in UbiquiTx, Inc. M.P.D. also has financial interests in Gauntlet, Inc. Glycobia, Inc., Resilience, Inc. and Versatope Therapeutics, Inc. M.P.D.’s and P.C.’s interests are reviewed and managed by Cornell University and Duke University, respectively, in accordance with their conflict-of-interest policies. All other authors declare no competing interests.
Databáze:	MEDLINE
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