Ex Vivo Expanded 3D Human Kidney Spheres Engraft Long Term and Repair Chronic Renal Injury in Mice

Harari‐Steinberg, Orit; Omer, Dorit; Gnatek, Yehudit; Pleniceanu, Oren; Goldberg, Sanja; Cohen-Zontag, Osnat; Pri‐Chen, Sara; Kanter, Itamar; Haim, Nissim; Becker, Eli; Ankawa, Roi; Fuchs, Yaron; Kalisky, Tomer; Dotan, Zohar; Dekel, Benjamin

doi:10.1016/j.celrep.2019.12.047

Cited by 27 publications

(63 citation statements)

References 63 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…were recovered from healthy margins of renal cell carcinoma (RCC) tumors, resected from partial or total nephrectomies 1,15 . Collected tissues were washed with cold HBSS (Invitrogen) and minced into ~ 1 mm cubes using sterile surgical scalpels followed by incubation at 37 °C for 2 h in Iscoves' Mod Dulbecco's Medium (IMDM) (Invitrogen) supplemented with 0.1% collagenase IV (Invitrogen).…”

Section: Establishment Of Primary Cultures From the Human Kidney Hummentioning

confidence: 99%

Human kidney clonal proliferation disclose lineage-restricted precursor characteristics

Cohen-Zontag

Gershon

Harari‐Steinberg

et al. 2020

Sci Rep

Self Cite

View full text Add to dashboard Cite

In-vivo single cell clonal analysis in the adult mouse kidney has previously shown lineage-restricted clonal proliferation within varying nephron segments as a mechanism responsible for cell replacement and local regeneration. To analyze ex-vivo clonal growth, we now preformed limiting dilution to generate genuine clonal cultures from one single human renal epithelial cell, which can give rise to up to 3.4 * 106 cells, and analyzed their characteristics using transcriptomics. A comparison between clonal cultures revealed restriction to either proximal or distal kidney sub-lineages with distinct cellular and molecular characteristics; rapidly amplifying de-differentiated clones and a stably proliferating cuboidal epithelial-appearing clones, respectively. Furthermore, each showed distinct molecular features including cell-cycle, epithelial-mesenchymal transition, oxidative phosphorylation, BMP signaling pathway and cell surface markers. In addition, analysis of clonal versus bulk cultures show early clones to be more quiescent, with elevated expression of renal developmental genes and overall reduction in renal identity markers, but with an overlapping expression of nephron segment identifiers and multiple identity. Thus, ex-vivo clonal growth mimics the in-vivo situation displaying lineage-restricted precursor characteristics of mature renal cells. These data suggest that for reconstruction of varying renal lineages with human adult kidney based organoid technology and kidney regeneration ex-vivo, use of multiple heterogeneous precursors is warranted.

show abstract

Section: Establishment Of Primary Cultures From the Human Kidney Hummentioning

confidence: 99%

Human kidney clonal proliferation disclose lineage-restricted precursor characteristics

Cohen-Zontag

Gershon

Harari‐Steinberg

et al. 2020

Sci Rep

Self Cite

View full text Add to dashboard Cite

show abstract

“…Therefore, it would be important to test ACE2 treatment and SARS-CoV-2 in three-dimensional kidney spheroids/organoids of adult origin harboring mature phenotypes. 36 Human soluble ACE2 1-740 is a large molecule of approximately 110 kD in size. 11,37 In fact, under native condition, ACE2 1-740 is even bigger, as it forms homodimers roughly double the size of the monomeric protein, 31 a process mediated by the collectrin-like domain at the C-terminal part of ACE2 (amino acids 616-726).…”

Section: Discussionmentioning

confidence: 99%

A Novel Soluble ACE2 Variant with Prolonged Duration of Action Neutralizes SARS-CoV-2 Infection in Human Kidney Organoids

Wysocki

Hassler

et al. 2021

JASN

118

View full text Add to dashboard Cite

BackgroundThere is an urgent need for approaches to prevent and treat SARS-CoV-2 infection. Administration of soluble ACE2 protein acting as a decoy to bind to SARS-CoV-2 should limit viral uptake mediated by binding to membrane-bound full-length ACE2, and further therapeutic benefit should result from ensuring enzymatic ACE2 activity to affected organs in patients with COVID-19.MethodsA short variant of human soluble ACE2 protein consisting of 618 amino acids (hACE2 1–618) was generated and fused with an albumin binding domain (ABD) using an artificial gene encoding ABDCon, with improved albumin binding affinity. Human kidney organoids were used for infectivity studies of SARS-CoV-2 in a BSL-3 facility to examine the neutralizing effect of these novel ACE2 variants.ResultsWhereas plasma ACE2 activity of the naked ACE2 1–618 and ACE2 1–740 lasted about 8 hours, the ACE2 1–618-ABD resulted in substantial activity at 96 hours, and it was still biologically active 3 days after injection. Human kidney organoids express ACE2 and TMPRSS2, and when infected with SARS-CoV-2, our modified long-acting ACE2 variant neutralized infection.ConclusionsThis novel ACE2 1–618-ABD can neutralize SARS-CoV-2 infectivity in human kidney organoids, and its prolonged duration of action should ensure improved efficacy to prevent viral escape and dosing convenience.

show abstract

“…Human adult kidney samples were recovered from healthy margins of renal cell carcinoma (RCC) tumors, resected from partial or total nephrectomies [1, 14]. Human Fetal Kidneys (hFKs) at gestational ages ranging from 15 to 22 weeks were harvested from fetuses that were electively aborted as previously described [15].…”

Section: Methodsmentioning

confidence: 99%

“…For both tissue engineering and direct cellular interventions, utilization of cell sources harboring bona a fide renal potential, e.g. renal lineage cells that function to generate or replace mature kidney cells, may prove beneficial [1].…”

Section: Introductionmentioning

confidence: 99%

Molecular characterization of clonal human renal forming cells

Osnat

Rotem

Orit

et al. 2020

Preprint

Self Cite

View full text Add to dashboard Cite

The adult kidney replaces its parenchyma in vivo in steady state and during regeneration by segment-specific clonal cell proliferation.To understand human adult kidney clonal cell growth, we derived tissue from human nephrectomies and performed limiting dilution to establish genuine clonal cultures from one single cell.Clonal efficiency of the human kidney was x%. Remarkably, a single renal cell could give rise to up to 3.3*10(6) cells. Phenotypically, two types of clonal cultures were apparent; a stably proliferating cuboidal epithelial-like appearing (EL) and a rapidly proliferating fibroblast-like appearing (FL). RNA sequencing of all clonal cultures separated FL from EL cultures according to proximal-distal/collecting renal epithelial tubular identity, respectively. Moreover, distinct molecular features in respect to cellcycle, epithelial-mesenchymal transition, oxidative phosphorylation, BMP signaling pathway and cell surface markers were observed for each clone type. Surprisingly, clonal expansion (>3 months) was sustained in EL clones harboring markers of mature kidney epithelia (high CD24, CDH1, EpCAM, EMA) in contrast to dedifferentiated FL clones (high NCAM1, serpine1), which showed fast lineage amplification and exhausted in a few weeks. Thus, the human adult kidney harbors progenitor cell function in which segment identity and the level of epithelial differentiation dictate clonal characteristics.2 4 . Liu, J., et al., Loss of SETD2 Induces a Metabolic Switch in Renal Cell Carcinoma Cell Lines toward Enhanced Oxidative Phosphorylation . J Proteome Res, 2019. 18(1): p. 331-340.2 5 . , L., et al., Clear Cell Renal Cell Carcinoma is linked to Epithelial-to-Mesenchymal Transition and to Fibrosis. Physiol Rep, 2017. 5(11.( 2 6 . Sanchez, D.J. and M.C. Simon, Genetic and metabolic hallmarks of clear cell renal cell carcinoma. Biochim Biophys Acta Rev Cancer, 2018. 1870(1): p. 23-31. Landolt2 7 .Buzhor, E., et al., Cell-based therapy approaches: the hope for incurable diseases. Regen Med, 2014. 9(5): p. 649-72. 2 8 . Chuang, C.H., et al., Molecular definition of a metastatic lung cancer state reveals a targetable CD109-Janus kinase-Stat axis. Nat Med, 2017. 23(3): p. 291-300.

show abstract

Ex Vivo Expanded 3D Human Kidney Spheres Engraft Long Term and Repair Chronic Renal Injury in Mice

Cited by 27 publications

References 63 publications

Human kidney clonal proliferation disclose lineage-restricted precursor characteristics

Human kidney clonal proliferation disclose lineage-restricted precursor characteristics

A Novel Soluble ACE2 Variant with Prolonged Duration of Action Neutralizes SARS-CoV-2 Infection in Human Kidney Organoids

Molecular characterization of clonal human renal forming cells

Contact Info

Product

Resources

About