Structural Variants of a Liver Fluke Derived Granulin Peptide Potently Stimulate Wound Healing

Abstract

Granulins are a family of growth factors involved in cell proliferation. The liver-fluke granulin, Ov-GRN-1, isolated from a carcinogenic liver fluke Opisthorchis viverrini, can significantly accelerate wound repair in vivo and in vitro. However, it is difficult to express Ov-GRN-1 in recombinant form at high yield, impeding its utility as a drug lead. Previously we reported that a truncated analogue ( Ov-GRN) promotes healing of cutaneous wounds in mice. NMR analysis of this analogue indicates the presence of multiple conformations, most likely as a result of proline cis/ trans isomerization. To further investigate whether the proline residues are involved in adopting the multiple confirmations, we have synthesized analogues involving mutation of the proline residues. We have shown that the proline residues have a significant influence on the structure, activity, and folding of Ov-GRN. These results provide insight into improving the oxidative folding yield and bioactivity of Ov-GRN and might facilitate the development of a novel wound healing agent.

Liver fluke granulin promotes exosome-mediated crosstalk and cellular microenvironment conducive to cholangiocarcinoma

“…The liver fluke paralogue, Ov-GRN-1, stimulates cell proliferation, wound healing, and angiogenesis in vitro and contributes to the pathogenesis of opisthorchiasis in vivo (17)(18)(19)(20)(21)(22). We have exploited this link infection to explore the role of the secreted liver fluke granulin in premalignant lesions by undertaking programmed CRISPR/Cas9 knockout of the Ov-GRN-1 gene from the genome of O. viverrini.…”

“…Whereas the full complement of metabolites released by this parasite remains generally to be investigated for roles of communication and disease, a secreted protein termed liver fluke granulin (Ov-GRN-1) has been the focus of increasing investigation. Ov-GRN-1 is a paralogue of human granulin, which like the human granulin stimulates cell proliferation and wound healing, and which appears to contribute to the pathogenesis of opisthorchiasis (17)(18)(19)(20)(21). We have exploited this link to more deeply explore the role of Ov-GRN-1 in pre-malignant lesions of the bile duct by undertaking programmed CRISPR/Cas9 knockout of the its cognate gene.…”

Defined Small Molecules Produced by Himalayan Medicinal Plants Display Immunomodulatory Properties

“…H69s were grown under similar conditions with growth factor-supplemented specialized complete media; DMEM/F12 with high glucose, 10% fetal bovine serum (FBS), 1× antibiotic/antimycotic, 25 µg/mL adenine, 5 µg/mL insulin, 1 µg/mL epinephrine, 8.3 µg/mL holo-transferrin, 0.62 µg/mL, hydrocortisone, 13.6 ng/mL T3 and 10 ng/mL EGF (Life Technologies). Cytotoxicity screening of the test compounds against the H69 cell line was performed using an xCELLigence SP system (ACEA Biosciences Inc., San Diego, CA, USA) as described by us previously [ 65 ]. All experiments were carried out as per the manufacturer’s instructions with 0.25–60 min read intervals using the real time cell assay (RTCA) software (ACEA Biosciences Inc., USA).…”

Harnessing helminth-driven immunoregulation in the search for novel therapeutic modalities

“…The liver fluke, Opisthorchis viverrini, secretes a granulin (GRN)-like growth factor, Ov-GRN-1, which promotes wound healing and angiogenesis [87]. Recombinant Ov-GRN-1 is challenging to express in scalable form, so a readily-synthesized bioactive peptide fragment of Ov-GRN-1 that retained both in vitro and in vivo wound healing properties was identified and retained the in vivo therapeutic properties of the parent protein [88]. Given the alarming incidence of T2D and associated comorbidities such as nonhealing diabetic foot ulcers, topical growth factor-like proteins and peptides from helminths address an area of great unmet need [89,90].…”