Whole Organism Profiling of the Timp Gene Family

Abstract

Tissue inhibitor of metalloproteinases (TIMPs/Timps) are an endogenous family of widely expressed matrisome-associated proteins that were initially identified as inhibitors of matrix metalloproteinase activity (Metzincin family proteases). Consequently, TIMPs are often considered simply as protease inhibitors by many investigators. However, an evolving list of new metalloproteinase-independent functions for TIMP family members suggests that this concept is outdated. These novel TIMP functions include direct agonism/antagonism of multiple transmembrane receptors, as well as interactions with matrisome targets. While the family was fully identified over 2 decades ago, there has yet to be an in-depth study describing the expression of TIMPs in normal tissues of adult mammals. An understanding of the tissues and cell-types that express TIMPs 1 through 4, in both normal and disease states are important to contextualize the growing functional capabilities of TIMP proteins, which are often dismissed as non-canonical. Using publicly available single cell RNA sequencing data from the Tabula Muris Consortium, we analyzed approximately 100,000 murine cells across nineteen tissues from non-diseased organs, representing seventy-three annotated cell types, to define the diversity in Timp gene expression across healthy tissues. We describe that Timp genes display unique expression profiles across tissues and organ-specific cell types. Within annotated cell-types, we identify clear and discrete cluster-specific patterns of Timp expression, particularly in cells of stromal and endothelial origins. Differential expression and gene set pathway analysis provide evidence of the biological significance of Timp expression in these identified cell sub-types, which are consistent with novel roles in normal tissue homeostasis and changing roles in disease progression. This understanding of the tissues, specific cell types and conditions of the microenvironment in which Timp genes are expressed adds important physiological context to the growing array of novel TIMP protein functions.

Timp2 loss-of-function mutation and TIMP2 treatment in murine model of NSCLC: modulation of immunosuppression and oncogenic signaling

“…Tissue inhibitors of metalloproteinases (TIMPs) are endogenous inhibitors of metalloproteinase (MP) activity, with TIMP2 displaying the broadest expression profile of the family ( 13, 14 ). In addition to their functions as inhibitors of MP activity, TIMPs demonstrate a growing list of MP-independent functions that include the regulation of growth factor signaling, cytokine-like signaling capabilities, and direct modulation of cell behavior ( 14–16 ).…”

“…Developing strategies include the normalization of the microenvironment to support anti-tumor immune cell functions, reduce extracellular matrix turnover, and restore homeostatic mechanisms that prevent tumor progression (11,12). Tissue inhibitors of metalloproteinases (TIMPs) are endogenous inhibitors of metalloproteinase (MP) activity, with TIMP2 displaying the broadest expression profile of the family (13,14). In addition to their functions as inhibitors of MP activity, TIMPs demonstrate a growing list of MPindependent functions that include the regulation of growth factor signaling, cytokine-like signaling capabilities, and direct modulation of cell behavior (14)(15)(16).…”

Extracellular Proximity Labeling Reveals an Expanded Interactome for the Matrisome Protein TIMP2

“…ePL reveals the TIMP2 proximal interactome in HT1080 cells HT1080 cells are an epithelial-like fibrosarcoma cell line that has been extensively utilized by researchers studying the biology of TIMPs and MPs [36][37][38]. Considering the strong correlation between TIMP2 expression and mesenchymal cells [39] (Figure 1B), they represent an ideal cell line for preliminary ePL studies. A total of 4 experiments were performed, representing duplicates from TurboID and BioID2 variations, fully summarized in Table S2.…”