A Novel Three–Dimensional Human Peritubular Microvascular System

Ligresti, Giovanni; Nagao, Ryan J.; Xue, Jing; Choi, Yoon Jung; Xu, Jin; Ren, Shen; Aburatani, Takahide; Anderson, Susan K.; MacDonald, James W.; Bammler, Theo K.; Schwartz, Stephen M.; Muczynski, Kimberly A.; Duffield, Jeremy S.; Himmelfarb, Jonathan; Zheng, Ying

doi:10.1681/asn.2015070747

Cited by 84 publications

(103 citation statements)

References 30 publications

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“…Because the glycocalyx composition is critically dependent upon shear, the cellular environment, and endothelial function, we used an experimental setup in which endothelial cells were exposed to laminar flow and cultured on top of pericytes, meant to mimic as closely as possible the in vivo situation. Endothelial cells show a remarkable heterogeneity throughout the vascular tree and may therefore differ in their response to injury (37,38). Despite this heterogeneity, HUVECs are capable of expressing heparanase (39), and in this model, adding DHS-thereby mimicking the diabetic milieu-increased endothelial heparanase expression.…”

Section: Discussionmentioning

confidence: 84%

Atrasentan Reduces Albuminuria by Restoring the Glomerular Endothelial Glycocalyx Barrier in Diabetic Nephropathy

et al. 2016

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Atrasentan, a selective endothelin A receptor antagonist, has been shown to reduce albuminuria in type 2 diabetes. We previously showed that the structural integrity of a glomerular endothelial glycocalyx is required to prevent albuminuria. Therefore we tested the potential of atrasentan to stabilize the endothelial glycocalyx in diabetic apolipoprotein E (apoE)-deficient mice in relation to its antialbuminuric effects. Treatment with atrasentan (7.5 mg/kg/day) for 4 weeks reduced urinary albumin-tocreatinine ratios by 26.0 6 6.5% (P < 0.01) in apoE knockout (KO) mice with streptozotocin-induced diabetes consuming an atherogenic diet, without changes in gross glomerular morphology, systemic blood pressure, and blood glucose concentration. Endothelial cationic ferritin surface coverage, investigated using large-scale digital transmission electron microscopy, revealed that atrasentan treatment increases glycocalyx coverage in diabetic apoE KO mice from 40.7 6 3.2% to 81.0 6 12.5% (P < 0.05). This restoration is accompanied by increased renal nitric oxide concentrations, reduced expression of glomerular heparanase, and a marked shift in the balance of M1 and M2 glomerular macrophages. In vitro experiments with endothelial cells exposed to laminar flow and cocultured with pericytes confirmed that atrasentan reduced endothelial heparanase expression and increased glycocalyx thickness in the presence of a diabetic milieu. Together these data point toward a role for the restoration of endothelial function and tissue homeostasis through the antialbuminuric effects of atrasentan, and they provide a mechanistic explanation for the clinical observations of reduced albuminuria with atrasentan in diabetic nephropathy.End-stage renal disease is inevitable in a majority of patients with diabetic nephropathy (1), despite optimal blood pressure treatment using drugs that interfere with the reninangiotensin system. Therefore there is a great need for additional strategies to slow the progression of chronic kidney disease in patients with diabetic nephropathy. One such strategy involves interaction with the endothelin (ET) system. Numerous studies involving experimental animal models have implicated ET in the pathogenesis of diabetic nephropathy (2). Moreover, clinical studies show promise for ET receptor antagonists in the treatment of diabetic nephropathy (3-6). This is particularly true for selective ET A receptor blockers; ET A receptor signaling seems to be involved in key renal pathophysiological processes such as the inflammatory response of renal epithelium to albumin (7), whereas stimulation of the associated concomitant ET B receptor can restore endothelial dysfunction by inducing endothelial nitric oxide (NO) production (8-10). Because actual loss of renal function is a late indicator of disease, albuminuria has been put forward as a sensitive surrogate marker for ongoing renal injury in diabetic nephropathy. In this respect ET A receptor blockers seem to have a striking

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Section: Discussionmentioning

confidence: 84%

Atrasentan Reduces Albuminuria by Restoring the Glomerular Endothelial Glycocalyx Barrier in Diabetic Nephropathy

et al. 2016

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“…In contrast, HUVECs, human cardiac MVECs or murine AECs had robust angiogenic responses in this assay (Fig 1D and S1H). To determine whether this finding was recapitulated in humans, we purified and expanded human KMVECs [37]. In the collagen invasion assay, human KMVECs similarly lacked angiogenic capacity (Fig 1E), even though human KMVECs expressed similar levels of FLT1 , KDR , NRP1 , and NRP2 in comparison to HUVECs (Fig S1D-G respectively).…”

Section: Resultsmentioning

confidence: 99%

“…Similar studies in mouse and human kidney disease, however, indicate this is not the case [50, 65, 66]. Although a mild angiogenic response occurs within a few days following acute kidney injury, all models of chronic kidney disease are ultimately characterized by vascular regression without significant vascular regeneration [38, 67]. Accumulating evidence indicates loss of the kidney microvasculature may be a central or early problem in many forms of kidney disease [55].…”

Section: Discussionmentioning

confidence: 99%

“…Mouse AECs were cultured in Complete Mouse Endothelial Cell Medium (CellBiologics). Human kidney MVECs were purified from adult or fetal kidneys (IRB447773EA University of Washington) and New England Medical Center (Boston, MA) respectively using methods as described [37, 38] with modifications. Human kidney MVECs were characterized as a CD31 (BD 555446, 1:50) positive, PDGFRβ (Biolegend 18A1, 1:200) and CD45 (Biolegend H130, 1:400) negative population.…”

Section: Methodsmentioning

confidence: 99%

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Hyperactive FOXO1 results in lack of tip stalk identity and deficient microvascular regeneration during kidney injury

et al. 2017

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Loss of the microvascular (MV) network results in tissue ischemia, loss of tissue function, and is a hallmark of chronic diseases. The incorporation of a functional vascular network with that of the host remains a challenge to utilizing engineered tissues in clinically relevant therapies. We showed that vascular-bed-specific endothelial cells (ECs) exhibit differing angiogenic capacities, with kidney microvascular endothelial cells (MVECs) being the most deficient, and sought to explore the underlying mechanism. Constitutive activation of the phosphatase PTEN in kidney MVECs resulted in impaired PI3K/AKT activity in response to vascular endothelial growth factor (VEGF). Suppression of PTEN in vivo resulted in microvascular regeneration, but was insufficient to improve tissue function. Promoter analysis of the differentially regulated genes in KMVECs suggests that the transcription factor FOXO1 is highly active and RNAseq analysis revealed that hyperactive FOXO1 inhibits VEGF-Notch-dependent tip-cell formation by direct and indirect inhibition of DLL4 expression in response to VEGF. Inhibition of FOXO1 enhanced angiogenesis in human bio-engineered capillaries, and resulted in microvascular regeneration and improved function in mouse models of injury-repair.

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“…Activation of these endothelial cells with TNF resulted in decreased flow and microchannel occlusion, whereas exposure of the cells to Shiga-like toxin 2 resulted in the formation of thrombi that occluded the microchannels, indicating that this design would be suitable for studying drug-induced thrombotic microangiopathy. More recently, a protocol has been developed for isolating human kidney peritubular microvascular endothelial cells (HKMECs) and culturing them in a microfluidic device 138 . These HKMECs have a markedly different transcriptional profile from that of human umbilical vein endothelial cells (HUVECs) cultured under the same conditions and showed lower angiogenic potential and increased responsiveness to flow, demonstrating that the kidney microvasculature has specific properties that might not be replicated in models that use other endothelial cell types or lack physiological features such as flow.…”

Section: Preclinical Screens Of Nephrotoxicitymentioning

confidence: 99%

Advances in predictive in vitro models of drug-induced nephrotoxicity

et al. 2018

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In vitro screens for nephrotoxicity are currently poorly predictive of toxicity in humans. Although the functional proteins that are expressed by nephron tubules and mediate drug susceptibility are well known, current in vitro cellular models poorly replicate both the morphology and the function of kidney tubules and therefore fail to demonstrate injury responses to drugs that would be nephrotoxic in vivo. Advances in protocols to enable the directed differentiation of pluripotent stem cells into multiple renal cell types and the development of microfluidic and 3D culture systems have opened a range of potential new platforms for evaluating drug nephrotoxicity. Many of the new in vitro culture systems have been characterized by the expression and function of transporters, enzymes, and other functional proteins that are expressed by the kidney and have been implicated in drug-induced renal injury. In vitro platforms that express these proteins and exhibit molecular biomarkers that have been used as readouts of injury demonstrate improved functional maturity compared with static 2D cultures and represent an opportunity to model injury to renal cell types that have hitherto received little attention. As nephrotoxicity screening platforms become more physiologically relevant, they will facilitate the development of safer drugs and improved clinical management of nephrotoxicants.

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A Novel Three–Dimensional Human Peritubular Microvascular System

Cited by 84 publications

References 30 publications

Atrasentan Reduces Albuminuria by Restoring the Glomerular Endothelial Glycocalyx Barrier in Diabetic Nephropathy

Atrasentan Reduces Albuminuria by Restoring the Glomerular Endothelial Glycocalyx Barrier in Diabetic Nephropathy

Hyperactive FOXO1 results in lack of tip stalk identity and deficient microvascular regeneration during kidney injury

Advances in predictive in vitro models of drug-induced nephrotoxicity

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