Table 2.
Preclinical strategies targeting lymphangiogenesis in chronic renal injury
| Strategy | Mechanism of Action and Model | Effect on Renal Lymphatics | Effect of Strategy on Model of CKD |
|---|---|---|---|
| VEGFC-156S | Mutated form of VEGF-C and selective agonist of VEGFR-3 to promote lymphangiogenesis.98 10 μg/d delivered IP over 14 d in murine UUO99 | Increase in cross-sectional area of LYVE-1+ total and perivascular renal lymphatics compared with control | Reduction in fibrotic remodeling (Sirius red stain) |
| Reduction in collagen I deposition (Western blot) | |||
| Reduction in infiltrating macrophages (IHC) | |||
| Reduction in TGF-β1 expression (Western blot) | |||
| Ksp-rtTA;TRE-Vegfd mice | Transgenic mice with doxycycline-dependent overexpression of VEGF-D from tubular epithelium to promote lymphangiogenesis. Tested in l-NAME–dependent hypertensive nephropathy for 2 wk with or without 3 wk high-salt diet100 | Increase in cross-sectional area of LYVE-1+ cortical renal lymphatics and branches per artery compared with control | Reduction in infiltrating macrophage and T lymphocytes with high-salt diet (flow cytometry) |
| Reduction in infiltrating macrophage and DCs without high-salt diet (flow cytometry) | |||
| Pod-rtTA;TRE-Vegfc mice | Transgenic mice with doxycycline-dependent overexpression of VEGF-C from podocytes. Diabetic nephropathy induced using 50 mg/kg STZ per day for 5 d.101 Doxycycline either given before or 4 wk after STZ injection | Effects attributed to glomerular VEGF activity, so renal lymphatics were not investigated | Reduction in urinary albumin-creatinine ratio (ELISA) |
| Reduction in mesangial matrix expansion (histology) | |||
| Reduction in collagen I (Sirius red stain) | |||
| IMC-3C5 | Anti–VEGFR-3 antibody delivered to inhibit lymphangiogenesis in rats with adriamycin nephropathy.102 At 6 wk after adriamycin treatment, 40 mg/kg body wt of IMC-C35 delivered three times per wk IP | Reduction in cross-sectional area of PDPN+ cortical lymphatics in both adriamycin-treated and non-adriamycin-treated kidneys compared with controls | No significant reduction in infiltrating macrophage or T lymphocytes (IHC) |
| No significant reduction in tubulointerstitial fibrosis (histology) | |||
| No significant reduction in collagen I (IHC and qPCR) | |||
| Lyve1-Cre;R26R-DTR mice | Transgenic mice expression DTR in LYVE-1+ cells and their progeny. Tested in murine UUO and IRI with 7 d follow-up after a single IP dose of 1.25 ng/kg body wt DT64 | Significant reduction in cross-sectional area of LYVE-1+ lymphatic vessels assessed at 3 d after DT administration | Reduction in DCs, macrophages, T and B lymphocytes, neutrophils, and NK cells in UUO (flow cytometry) |
| Reduction of inflammatory cytokines in UUO (qPCR) | |||
| Reduction in fibrosis (qPCR, IHC, and Sirius red) | |||
| LYVE-1 or VEGFR-3 soluble fusion proteins | Soluble LYVE-1 or VEGFR-3 proteins hypothesized to inhibit lymphangiogenesis through sequestering lymphangiogenic growth factors. Injected via tail vein before or after UUO or IRI induction in mice64 | Significant reduction in cross-sectional area of LYVE-1+ lymphatic vessels assessed at 7 d after UUO surgery | Reduction in DCs and T lymphocytes in UUO kidneys (flow cytometry) |
| Reduction of inflammatory cytokines in UUO (qPCR) | |||
| Reduction in fibrosis (qPCR, IHC, and Sirius red) |
IP, intraperitoneal; IHC, immunohistochemistry; l-NAME, nitro- l -arginine methyl ester; STZ, streptozocin; qPCR, quantitative PCR; DTR, diphtheria toxin receptor; DT, diphtheria toxin; NK, natural killer.