In depth quantification of extracellular matrix proteins from human pancreas

ABSTRACT: Extracellular matrix (ECM) is an important component of the pancreatic microenvironment which regulates β cell proliferation, differentiation and insulin secretion. Protocols have recently been developed for the decellularization of the human pancreas to generate functional scaffolds and hydrogels. In this work, we characterized human pancreatic ECM composition before and after decellularization using isobaric dimethylated leucine (DiLeu) labeling for relative quantification of ECM proteins. A novel correction factor was employed in the study to eliminate the bias introduced during sample preparation. In comparison to the commonly employed proteomic approaches (urea and FASP), a recently developed surfactant and chaotropic agent assisted sequential extraction/on pellet digestion (SCAD) protocol was proven to be a superior strategy for ECM protein extraction of human pancreatic ECM matrix. The quantitative proteomic results revealed the preservation of matrisome proteins while most of the cellular proteins were removed. This method was compared with a well-established label-free quantification (LFQ) approach which rendered similar expressions of different categories of proteins (collagens, ECM glycoproteins, proteoglycans, etc.). The distinct expression of ECM proteins was quantified comparing adult and fetal pancreas ECM, shedding light on the correlation between matrix composition and post-natal β cell maturation. Despite the distinct profiles of different subcategories in the native pancreas, the distribution of matrisome protein exhibited similar trends after the decellularization process. Our method generates the largest dataset of matrisome proteins from a single tissue type. These results provide valuable insight into the possibilities of constructing a bioengineered pancreas. It also facilitates an understanding of the significant roles that matrisome proteins play in post-natal cell maturation.

INSTRUMENT(S): Orbitrap Fusion Lumos

ORGANISM(S): Homo Sapiens (human)

TISSUE(S): Pancreas

SUBMITTER: Fengfei Ma  

LAB HEAD: Lingjun Li

PROVIDER: PXD012460 | Pride | 2019-06-17

REPOSITORIES: Pride