Simultaneous targeting of CD44 and MMP9 catalytic and hemopexin domains as a therapeutic strategy

Abstract

Crosstalk of the oncogenic matrix metalloproteinase-9 (MMP9) and one of its ligands, CD44, involves cleavage of CD44 by the MMP9 catalytic domain, with the CD44–MMP9 interaction on the cell surface taking place through the MMP9 hemopexin domain (PEX). This interaction promotes cancer cell migration and invasiveness. In concert, MMP9-processed CD44 induces the expression of MMP9, which degrades ECM components and facilitates growth factor release and activation, cancer cell invasiveness, and metastasis. Since both MMP9 and CD44 contribute to cancer progression, we have developed a new strategy to fully block this neoplastic process by engineering a multi-specific inhibitor that simultaneously targets CD44 and both the catalytic and PEX domains of MMP9. Using a yeast surface display technology, we first obtained a high-affinity inhibitor for the MMP9 catalytic domain, which we termed C9, by modifying a natural non-specific MMP inhibitor, N-TIMP2. We then conjugated C9 via a flexible linker to PEX, thereby creating a multi-specific inhibitor (C9-PEX) that simultaneously targets the MMP9 catalytic and PEX domains and CD44. It is likely that, via its co-localization with CD44, C9-PEX may compete with MMP9 localization on the cell surface, thereby inhibiting MMP9 catalytic activity, reducing MMP9 cellular levels, interfering with MMP9 homodimerization, and reducing the activation of downstream MAPK/ERK pathway signaling. The developed platform could be extended to other oncogenic MMPs as well as to other important target proteins, thereby offering great promise for creating novel multi-specific therapeutics for cancer and other diseases.

MMP9: A Tough Target for Targeted Therapy for Cancer

“…The strategy relied on the naturally occurring inhibitor TIMP2 since it binds more strongly to MMP9 than to other MMPs. The mechanism of action of C9-PEX, through which it antagonizes the interaction of MMP9 with CD44, leads to the suppression of ECM degradation and subsequently cancer cell migration and invasion [ 170 ]. Even though many such synthetic small compounds with increased selectivity have been developed [ 171 , 172 ], their clinical utility has not yet been determined, and there are currently no MMP inhibitors (MMPIs) in clinical trials for cancer.…”

Protective Mechanism of Ethyl Gallate against Intestinal Ischemia‐Reperfusion Injury in Mice by in Vivo and in Vitro Studies Based on Transcriptomics

“…Matrix metalloproteinases (MMPs) were a class of zinc‐dependent proteases that were abundant in cancerous and inflammatory tissues and whose main function was to degrade and remodel the ECM [39–41] . MMP9 is a type of gelatinase whose main function is to degrade type IV collagen, modulate adhesion factor activity, and degrade and disrupt basement membrane (BM) structure, leading to degradation and remodeling of the ECM, and promoting migration and invasion of cancer cells [42,43] . In intestinal tissues, MMP9 promoted ECM degradation and inflammation, leading to disruption of intestinal barrier function and structure, which in turn triggered intestinal tissue damage [22,38] .…”

“…[39][40][41] MMP9 is a type of gelatinase whose main function is to degrade type IV collagen, modulate adhesion factor activity, and degrade and disrupt basement membrane (BM) structure, leading to degradation and remodeling of the ECM, and promoting migration and invasion of cancer cells. [42,43] In intestinal tissues, MMP9 promoted ECM degradation and inflammation, leading to disruption of intestinal barrier function and structure, which in turn triggered intestinal tissue damage. [22,38] The sequencing results indicated that EG might inhibit the degradation and remodeling of ECM by regulating the expression of MMP9.…”

Ozanimod Attenuates Human Cerebrovascular Endothelial Derived MMP-9 Activity and Preserves Barrier Properties Following In Vitro Acute Ischemic Injury

“…The copyright holder for this preprint this version posted February 3, 2023. ; https://doi.org/10.1101/2023.02.01.526738 doi: bioRxiv preprint endothelial barrier function, (11) proliferation and survival, (73) and as an anchor point for MMP-9. (17,74) In this study we utilized an immunocytochemical approach to ascertain the effects of HGD exposure as well as treatment with ozanimod on the levels and nuclear overlap of CD44. Figure 7A demonstrates representative immunocytochemical images of HBMECs with anti-CD44 (green) and DAPI stained nuclei (blue) following normoxia or HGD exposure +/-ozanimod.…”

“…In addition to tight junctions, the endothelium within the neurovascular unit has been shown to secrete ECM which plays an integral role in maintaining vessel and BBB integrity. [Reviewed in (12)] During the progression of AIS, tight junctions (13,14,15,16,17) and ECM are disrupted (18,19,20,21,22), which in part can be contributed to the notable acute activation of MMP-9. (23,24,25,26) Moreover, MMP-9 is postulated to be one of the drivers responsible for the biphasic manner in which the BBB is disrupted, where the acute onset phase is classified as detrimental, and the late onset is beneficial in promoting vascular (which was not certified by peer review) is the author/funder.…”