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bioRxiv · 10.1101/2025.05.22.655451

Mutations in specific MT5-MMP domains prevent the accumulation of toxic APP metabolites and serve as templates for peptide-based therapeutics in cell models of Alzheimer's disease

Rivera, S.·Belio-Mairal, P.·Kamitsou, A.·Stephan, D.·Jullien, N.·Thiane, D.·Ramos, M.·Louis, L.·Benoist, F.·Serrano, B.·David, M.·KHRESTCHATISKY, M.·Lecorche, P.·Nivet, E.
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Abstract

Our prior work established a pivotal role for membrane-type 5-matrix metalloproteinase (MT5-MMP) in Alzheimers disease (AD) pathogenesis, particularly through its C-terminal transmembrane (TM) and intracellular (IC) domains, which influence the fate of major toxic metabolites of toxic amyloid precursor protein (APP) metabolites, particularly C99 and A{beta}. Hypothesizing that modifications in these domains could modulate C99 and A{beta} levels, we engineered MT5-MMP variants with deletions or substitutions in the TM/IC domains or specific IC amino acid clusters. When co-transfected into human cell lines accumulating C99, certain IC domain mutations promoted C99 degradation and reduced A{beta} levels, while other mutations had divergent effects. High content imaging further revealed that MT5-MMP IC domain modification altered C99 subcellular trafficking within the endomembrane system, impacting its processing. Proximity ligation assays confirmed the importance of the IC domain in MT5-MMP colocalization and potential interaction with C99. To translate these findings, we synthetized peptides mimicking the MT5-MMP IC domain, incorporating mutations that reduce C99 and/or A{beta} levels. One peptide effectively lowered C99 levels in an in vitro AD model. Overall, this study highlights the importance of specific amino acids in the C-terminal domains of MT5-MMP for regulating C99 and A{beta} metabolism. It also provides new insights for developing MT5-MMP-based therapeutic strategies against AD, exploiting the unique properties of specific mutations in MT5-MMP to prevent toxic accumulation of C99 and A{beta}. O_FIG O_LINKSMALLFIG WIDTH=200 HEIGHT=86 SRC="FIGDIR/small/655451v3_ufig1.gif" ALT="Figure 1"> View larger version (27K): org.highwire.dtl.DTLVardef@a6b04aorg.highwire.dtl.DTLVardef@1800721org.highwire.dtl.DTLVardef@542505org.highwire.dtl.DTLVardef@4a6d7b_HPS_FORMAT_FIGEXP M_FIG O_FLOATNOGRAPHICAL ABSTRACTC_FLOATNO C_FIG

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Rivera, S., Belio-Mairal, P., Kamitsou, A., Stephan, D., Jullien, N., Thiane, D., Ramos, M., Louis, L., Benoist, F., Serrano, B., David, M., KHRESTCHATISKY, M., Lecorche, P., Nivet, E.. 2025-05-23. Mutations in specific MT5-MMP domains prevent the accumulation of toxic APP metabolites and serve as templates for peptide-based therapeutics in cell models of Alzheimer's disease. https://doi.org/10.1101/2025.05.22.655451

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