Recombinant VEGF-C (Cys156Ser) improves mesenteric lymphatic drainage and gut immune surveillance in experimental cirrhosis

Pinky Juneja; Syed Nazrin Ruhina Rahman; Deepika Jakhar; Akash Kumar Mourya; Dinesh M Tripathi; Impreet Kaur; Vaibhav Tiwari; Sumati Rohilla; Abhishek Gupta; Preety Rawal; Sukriti Baweja; Archana Rastogi; V G M Naidu; Shiv K Sarin; Subham Banerjee; Savneet Kaur

doi:10.1016/j.jhepr.2023.100816

Recombinant VEGF-C (Cys156Ser) improves mesenteric lymphatic drainage and gut immune surveillance in experimental cirrhosis

JHEP Rep. 2023 Jun 17;5(10):100816. doi: 10.1016/j.jhepr.2023.100816. eCollection 2023 Oct.

Authors

Pinky Juneja¹, Syed Nazrin Ruhina Rahman², Deepika Jakhar¹, Akash Kumar Mourya¹, Dinesh M Tripathi¹, Impreet Kaur¹, Vaibhav Tiwari¹, Sumati Rohilla¹, Abhishek Gupta¹, Preety Rawal¹, Sukriti Baweja¹, Archana Rastogi³, V G M Naidu⁴, Shiv K Sarin⁵, Subham Banerjee², Savneet Kaur¹

Affiliations

¹ Department of Molecular and Cellular Medicine, Institute of Liver and Biliary Sciences, New Delhi, India.
² Department of Pharmaceutics, National Institute of Pharmaceutical Education and Research (NIPER)-Guwahati, Changsari, India.
³ Department of Pathology, Institute of Liver and Biliary Sciences, New Delhi, India.
⁴ Department of Pharmacology and Toxicology, NIPER-Guwahati, Changsari, India.
⁵ Department of Hepatology, Institute of Liver and Biliary Sciences, New Delhi, India.

Abstract

Background & aims: Lymphatic vessels (LVs) are crucial for maintaining abdominal fluid homoeostasis and immunity. In cirrhosis, mesenteric LVs (mLVs) are dilated and dysfunctional. Given the established role of vascular endothelial growth factor-C (VEGF-C) in improving LVs, we hypothesised that VEGF-C treatment could ameliorate the functions of mLVs in cirrhosis.

Methods: In this study, we developed a nanoformulation comprising LV-specific growth factor, recombinant human VEGF-C (Cys156Ser) protein (E-VEGF-C) and delivered it orally in different models of rat cirrhosis to target mLVs. Cirrhotic rats were given nanoformulation without VEGF-C served as vehicles. Drainage of mLVs was analysed using tracer dye. Portal and systemic physiological assessments and computed tomography were performed to measure portal pressures and ascites. Gene expression and permeability of primary mesenteric lymphatic endothelial cells (LyECs) was studied. Immune cells in mesenteric lymph nodes (MLNs) were quantified by flow cytometry. Endogenous and exogenous gut bacterial translocation to MLNs was examined.

Results: In cirrhotic rats, mLVs were dilated and leaky with impaired drainage. Treatment with E-VEGF-C induced proliferation of mLVs, reduced their diameter, and improved functional drainage. Ascites and portal pressures were significantly reduced in E-VEGF-C rats compared with vehicle rats. In MLNs of E-VEGF-C animals, CD8⁺CD134⁺ T cells were increased, whereas CD25⁺ regulatory T cells were decreased. Both endogenous and exogenous bacterial translocation were limited to MLNs in E-VEGF-C rats with reduced levels of endotoxins in ascites and blood in comparison with those in vehicle rats. E-VEGF-C treatment upregulated the expression of vascular endothelial-cadherin in LyECs and functionally improved the permeability of these cells.

Conclusions: E-VEGF-C treatment ameliorates mesenteric lymph drainage and portal pressure and strengthens cytotoxic T-cell immunity in MLNs in experimental cirrhosis. It may thus serve as a promising therapy to manage ascites and reduce pathogenic gut bacterial translocation in cirrhosis.

Impact and implications: A human recombinant pro-lymphangiogenic growth factor, VEGF-C, was encapsulated in nanolipocarriers (E-VEGF-C) and orally delivered in different models of rat liver cirrhosis to facilitate its gut lymphatic vessel uptake. E-VEGF-C administration significantly increased mesenteric lymphatic vessel proliferation and improved lymph drainage, attenuating abdominal ascites and portal pressures in the animal models. E-VEGF-C treatment limited bacterial translocation to MLNs only with reduced gut bacterial load and ascitic endotoxins. E-VEGF-C therapy thus holds the potential to manage ascites and portal pressure and reduce gut bacterial translocation in patients with cirrhosis.

Keywords: Liver cirrhosis; Lymphangiogenesis; Mesenteric lymphatic vessels; Portal hypertension; Targeted nano-delivery.