Urinary Kidney Biomarker Panel Detects Preclinical Antisense Oligonucleotide-Induced Tubular Toxicity

Abstract

Sensitive kidney safety assessment is important for successful drug development in both preclinical and clinical stages. The Food and Drug Administration recently qualified a composite measure of 6 urine creatinine-normalized biomarkers, such as clusterin, cystatin C, kidney injury molecule 1 (KIM-1), N-acetyl-β-d-glucosaminidase, neutrophil gelatinase-associated lipocalin (NGAL), and osteopontin, for monitoring kidney toxicity in early clinical trials. The qualification was based on small molecule drugs in humans, and the full panel has not been assessed in other species or for other drug modalities. This study evaluated the effects on these biomarkers for a constrained ethyl antisense oligonucleotide (tool ASO) with demonstrated kidney toxicity in mice compared to a control ASO of the same chemistry. Dosing 50 mg/kg of the tool ASO resulted in mild proximal tubular pathology and elevations in KIM-1, clusterin, NGAL, and cystatin C. A lower dose resulted in milder histopathology and lower biomarker increases. Unexpectedly, the control ASO induced mild elevations in KIM-1, NGAL, and cystatin C, despite the lack of pathology. Both KIM-1 and clusterin were most closely associated with kidney pathology and increased with the severity of injury. Altogether, our data suggest that a biomarker panel is a sensitive tool for the detection of preclinical ASO-induced kidney pathology.

Antibody-oligonucleotide conjugate achieves CNS delivery in animal models for spinal muscular atrophy

“…While the activity of the PMO was lowest in the kidney, the PMO concentration was the highest of all tissues evaluated, indicating a non-functional accumulation of PMO or antibody-PMO (Figure 4H). However, the high PMO accumulation did not result in enhanced levels of kidney injury molecule-1 (KIM-1), a marker of kidney injury previously observed in animals treated with high concentrations of oligonucleotides (Supplemental Figure S3) (36).…”

Safety, Tissue Distribution, and Metabolism of LNA-Containing Antisense Oligonucleotides in Rats

“…Six of these biomarkers (KIM-1, B2M, CYST, NGAL, OPN, and CLST) have been recently assessed for ASO-induced tubular toxicity in a mouse model, and 5 of them (KIM-1, B2M, CYST, NGAL, and CLST) resulted in treatment-related elevations associated with proximal tubular pathology, confirming their value as kidney injury biomarkers. 47…”

“…Six of these biomarkers (KIM-1, B2M, CYST, NGAL, OPN, and CLST) have been recently assessed for ASO-induced tubular toxicity in a mouse model, and 5 of them (KIM-1, B2M, CYST, NGAL, and CLST) resulted in treatment-related elevations associated with proximal tubular pathology, confirming their value as kidney injury biomarkers. 47 To our knowledge, only 2 recent studies on rat eyes and mouse kidneys have so far successfully detected oligonucleotides in tissue sections by MALDI MSI, 22,23 with approaches based on a time of flight detector. In the present study, an FTICR high mass-resolving analyzer was applied for the first time to ASO imaging, leading to major improvement in The LNA-containing ASO were observed in the kidney in proximal tubular epithelial cells as electron-dense gold-positive vesicles (EGPVs) with irregular shape consistent with endosomes or secondary lysosomes (A-D).…”

Assessing the fidelity of translation of non‐clinical assays: a Pharma perspective

“…In the non‐clinical literature, the utility of combining multiple studies is typically discussed only in more general terms (e.g., the integrated risk assessments in the International Conference on Harmonization S7B Guidance Document—Non‐clinical evaluation of the potential for delayed ventricular repolarization (QT interval prolongation) by human pharmaceuticals [ICH S7B] guidance document), though some examples of multi‐parametric assays have been published (e.g., for in vitro assessments of cardiotoxicity, Doherty et al, 2013; Sirenko et al, 2013). Other examples considering multiple assays include logistic and original linear regression models such as used recently to assess the utility of seven predictors of pro‐arrhythmic risk of drugs tested in repolarization studies with human‐induced pluripotent stem cell‐derived cardiomyocytes (Blinova et al, 2018) and a composite measure (consisting of six serum biomarkers for humans; FDA, 2021) predicting drug‐induced kidney toxicity in mice, recently published by Sandelius et al (2020).…”