Kidney Week

Abstract: SA-PO455

Mild AKI in a Murine Polymicrobial Sepsis Model and Its Value for Preclinical Testing of Human Cell Therapies

Session Information

• Development and Regenerative Medicine
  November 09, 2019 | Location: Exhibit Hall, Walter E. Washington Convention Center
  Abstract Time: 10:00 AM - 12:00 PM

Category: Development, Stem Cells, and Regenerative Medicine

• 500 Development, Stem Cells, and Regenerative Medicine

Authors

• Fazekas, Barbara, National University of Ireland Galway, Galway, Ireland

Barbara Fazekas,

• Alagesan, Senthilkumar, Orbsen Therapeutics Ltd, Galway, Ireland, Galway, Ireland

Senthilkumar Alagesan,

• Watson, Luke, Orbsen Therapeutics Ltd, Galway, Ireland, Galway, Ireland

Luke Watson,

• Ng, Olivia, Orbsen Therapeutics Ltd, Galway, Ireland, Galway, Ireland

Olivia Ng,

• Conroy, Callum Michael, Orbsen Therapeutics Ltd, Galway, Ireland, Galway, Ireland

Callum Michael Conroy,

• Garcia, Yolanda, National University of Ireland Galway, Galway, Ireland

Yolanda Garcia,

• Elliman, Stephen Joseph, Orbsen Therapeutics Ltd, Galway, Ireland, Galway, Ireland

Stephen Joseph Elliman,

• Griffin, Matthew D., National University of Ireland Galway, Galway, Ireland

Matthew D. Griffin,

Background

Even mild AKI greatly worsens the prognosis of sepsis. Allogeneic mesenchymal stem cells (allo-MSC) have potential therapeutic value in sepsis-associated (SA)-AKI but their optimal administration is undetermined. Improved analysis of human MSC effects in pre-clinical models of SA-AKI could improve future clinical trial design. We aimed to develop a model of mild SA-AKI in which to test a human allo-MSC cell product.

Methods

Caecal ligation and puncture (CLP) or Sham surgery with frequent post-procedural monitoring were performed in male C57BL/6 mice. Blood was sampled at 24, 48 and 72 hrs and kidney tissue collected at 72 hrs. Antibody-selected human umbilical cord (UC)-MSC were injected IV at 10⁶ cells/animal 4 hours post-CLP. Plasma and renal tubular neutrophil gelatinase-associated lipocalin (NGAL) were quantified by ELISA and IHC. Renal lymphoid and myeloid cells were quantified by multi-color flow cytometry of collagenase/DNAase-digested kidney.

Results

CLP was associated with 10-15% body weight loss and low (10%) mortality by 72 hrs. Plasma liver enzymes were mildly increased in CLP vs Sham but BUN and creatinine were not raised. Plasma (p)NGAL was markedly increased in CLP versus SHAM animals (p<0.001), peaking at 24 hrs and remaing elevated at 48 & 72 hrs. NGAL staining intensity in renal tubular epithelial cells strongly discriminated between CLP and SHAM at 72 hrs (Mean score 2.6±1.2 vs. 0.0±0.0, p=0.0003) and correlated with pNGAL. Intra-renal immune cell profiling indicated that SA-AKI was associated with proportionatley reduced T-cells, increased neutrophils and a complex modulation of MHC II⁺ mononuclear phagocytes (MP). A single administration of UC-MSC (compared to saline) resulted in less body weight loss (6.8±3.7% vs. 10.0±5.2%, p<0.05), trends toward lower pNGAL and renal NGAL staining intensity and reversal of SA-AKI-associated alterations to intra-renal T-cells, neutrophls and MP.

Conclusion

Mouse CLP with frequent post-procedural monitoring resulted in mild SA-AKI best detected by pNGAL, renal tubular NGAL staining intensity and altered intra-renal immune cell repertoire. The model allowed for quantification of the kidney-specific effects of an investigational human allo-MSC product in the setting of subclinical SA-AKI.

Funding

• Commercial Support – Orbsen Therapeutics Ltd.