VEGFR3 tyrosine kinase inhibition aggravates cisplatin nephrotoxicity

Abstract

Expansion of renal lymphatic networks, or lymphangiogenesis (LA) is well-recognized during development, and is now being implicated in kidney diseases. Though LA is associated with multiple pathological conditions, very little is known about its role in acute kidney injury (AKI). The purpose of this study was to evaluate the role of LA in a model of cisplatin-induced nephrotoxicity. LA is predominately regulated by VEGF-C and VEGF-D, ligands that exert their function through their cognate receptor VEGFR3. We demonstrate the use of MAZ51, a selective VEGFR3 inhibitor, which caused significantly worse structural and functional kidney damage in cisplatin nephrotoxicity. Apoptotic cell death and inflammation were also increased in the MAZ51 treated animals compared to vehicle-treated animals following cisplatin administration. Notably, MAZ51 caused significant upregulation of intrarenal phospho-NF-kB, phospho-JNK, and IL-6. Cisplatin nephrotoxicity is associated with vascular congestion due to endothelial dysfunction. Using three-dimensional tissue cytometry, a novel approach to explore lymphatics in the kidney, we detected significant vascular autofluorescence attributed to erythrocytes in cisplatin alone treated animals. Interestingly, no such congestion was detected in MAZ51 treated animals. We found increased renal vascular damage in MAZ51 treated animals, whereby MAZ51 caused a modest decrease in endothelial markers Emcn and vWF, with a modest increase in VEGFR2. Our findings identify a protective role for de novo LA in cisplatin nephrotoxicity and provide a rationale for the development of therapeutic approaches targeting LA. Our studies also suggest off target effects of MAZ51 on the vasculature in the setting of cisplatin nephrotoxicity.

Hydrogen Sulfide Attenuates Lymphedema Via the Induction of Lymphangiogenesis Through a PI3K/Akt‐Dependent Mechanism

“… 27 For instance, it is known that the insufficient formation or dysfunction of lymphatic vessels exacerbates pathological conditions. 19 , 21 , 28 Conversely, lymphangiogenesis can accelerate wound healing and subsequently maintain organ functions by improving inflammation and inhibiting fibrosis during the extreme phase of pathological conditions. 19 , 21 , 29 , 30 Thus, therapeutic lymphangiogenesis is a promising strategy for promoting reparative lymphangiogenesis of endogenous ecological responses to improve pathological conditions, such as ischemic heart disease, critical ischemic limb, and lymphedema.…”

Integrated cytometry with machine learning applied to high-content imaging of human kidney tissue for in-situ cell classification and neighborhood analysis

“…We hypothesize that these immune cell subpopulations may reflect different stages of activity and infiltration: from margination, extravasation and exiting vessels towards forming foci of inflammation within the peritubular space. These findings are only proof-of-principle of the capabilities of VTEA in using imaging-based data for discovery of spatially based cell niches and require further validation to fully determine biological relevance and generalizability 4,19 .…”

“…Antibody Conjugation and Validation. 14 of the 23 antibodies used here were conjugated in-house using the protocol outlined by Akoya Biosciences (Table 3) 19,20 . To conjugate barcodes to antibodies, antibodies were reduced using a "Reduction Master Mix" (Akoya Biosciences) to which lyophilized barcodes resuspended in molecular biology grade water and "Conjugation Solution" (Akoya Biosciences) were added and incubated for 2 hours at room temperature.…”

“…Since the initial description of V olumetric T issue E xploration and A nalysis (VTEA) as an open-source project, several excellent tools for tissue cytometry have been developed ( Table 1 ). In this time VTEA (v0.5.2-v0.7) has been used in several projects involving single imaging fields to large mesoscale multi-fluorescence kidney image volumes to perform supervised cytometry analysis with gating 4,5,19–24 . However, machine learning, data visualization and analysis tools that are useful for multidimensional, big-data scale imaging data were not previously implemented in VTEA’s uniquely integrated pipeline.…”

“…Since the initial description of Volumetric Tissue Exploration and Analysis (VTEA) as an open-source project, several excellent tools for tissue cytometry have been developed (Table 1). In this time VTEA (v0.5.2-v0.7) has been used in several projects involving single imaging fields to large mesoscale multifluorescence kidney image volumes to perform supervised cytometry analysis with gating 4,5,[19][20][21][22][23][24] .…”