Q CAO1, H YI1, A J. GILL2, M FOLEY3,4, C HUANG1, X CHEN1, C A. POLLOCK1
1Kolling Institute of Medical Research,Royal North Shore Hospital, Sydney Medical School, University of Sydney, St Leonards, Sydney, Australia, 2Department of Anatomical Pathology, NSW Health Pathology, Royal North Shore Hospital, St Leonards, Australia, 3AdAlta Pty. Ltd., 15/2 Park Dr., Bundoora, Australia, 4Department of Biochemistry and Genetics, La Trobe Institute for Molecular Science, La Trobe University, , Melbourne, Australia
Aim: To define the role of i-body AD-214 in renal fibrosis.
Background: Tissue fibrosis is the common pathological pathway in progressive chronic kidney disease (CKD). Current clinical practices are ineffective in limiting renal fibrosis. CXCR4 has been demonstrated to be central to the development of fibrosis. I-body AD-214, a redesigned form of i-body AD-114, is an Fc-Fusion protein that contains two AD-114 i-body molecules that bind with high affinity to CXCR4. The Fc Fragment of AD-214 greatly extends its half-life and therefore efficacy. AD-214 has been shown to be effective in limiting lung fibrosis. However, the role of AD-214 in renal fibrosis has not been investigated.
Methods: PTC cells were incubated with TGF-β1 (2ng/ml) with/without AD-214 (1μM or 2μM) for 48 hours. Supernatant was collected and collagen 3 (Col 3) and collagen 4 (Col 4) were measured by Western blot. Mice with unilateral ureteral obstruction (UUO) were administrated AD-214 every two days starting from one day after UUO for 14 days. Changes in renal morphology were examined by H&E staining. Renal histology, mRNA, analysed by qRT-PCR and extracellular matrix (ECM) protein expression detected by immunohistochemistry (IHC) were examined.
Results: AD-214 (2µM) suppressed TGF-β1-induced overexpression of Col 3 and Col 4 compared to a negative control i-body (P<0.05, n=4). In UUO model, AD-214 markedly ameliorated fibrotic kidney remodeling (P<0.05) and attenuated the UUO-induced overexpression of ECM including fibronectin (P<0.05) and collagens (P<0.05) in kidneys compared to negative control i-body (n=6-8).
Conclusions: Blocking CXCR4 using the i-body AD-214 is a promising therapeutic strategy to prevent the development of CKD.
Dr Qinghua Cao is an early-career researcher in Renal Medicine Group at Kolling Institute of Medical Research, University of Sydney. In the first year of her PhD study, Dr Cao has initially developed the collaboration with a pharmaceutical company, which then contributed to the success of a NHMRC application. Despite an early career, Dr Cao has published 12 papers and 12 conference abstracts in total. Among these publications, the review article in FASEB BioAdvances was regarded as TOP DOWNLOADED PAPER 2018-2019 and the abstract to 2018APCN obtained the best abstract award. Currently, she is working on multiple projects supported by NHMRC and different commercial partners.