# MITOCHONDRIAL RESPIROMETRY IN FROZEN BIOLOGICAL SAMPLES

> **NIH NIH R41** · ENSPIRE BIO, INC. · 2020 · $225,000

## Abstract

Abstract
 Impaired mitochondrial respiration plays a key role in metabolic, cardiovascular, and aging-related
diseases. However, mitochondrial respirometry analysis currently requires processing of the living tissue
sample within an hour after being taken from the patient. This requirement makes respirometry analysis largely
unfeasible to standard clinical practice and clinical studies. We invented a new technology to assess maximal
mitochondrial respiratory capacity in previously-frozen biological samples, which thus far been considered
impossible by the scientific community. Side-by-side comparison confirmed that our assay accurately reflects
results measured in fresh samples. Our technology thus overcomes the fundamental limitation of standard
approaches that required living tissue. Moreover, our approach consists of simplified sample preparation which
uses significantly less biological material.
 Enspire Bio will further develop this patented Frozen Mitos technology to enable respirometry
measurements for clinical applications and frozen samples, which currently is not available. We will achieve
our goals by Aim 1: Establishing standard protocols for oxygen consumption measurements from frozen tissue
samples derived from human and mouse tissue samples. We will further improve throughput by enabling
measurements of oxygen consumption from tissue homogenate instead of isolated mitochondria. Aim 2:
Establishing proprietary kits and delivering those to five beta testing sites including opinion-leading academic
labs, mitochondrial disease centers and a biotech and pharmaceutical company to test and provide feedback.
To provide frozen sample analysis to clients that have no capacity for wet lab assays, we will develop a CRO-
type service in-house where samples will be shipped to EnspireBio for analysis. Working with a set of pilot
academic, clinical and industry clients will provide feedback to establish procedures and the economics of the
CRO model approach for the frozen tissue respirometry.

## Key facts

- **NIH application ID:** 10011475
- **Project number:** 1R41ES031043-01A1
- **Recipient organization:** ENSPIRE BIO, INC.
- **Principal Investigator:** Orian S Shirihai
- **Activity code:** R41 (R01, R21, SBIR, etc.)
- **Funding institute:** NIH
- **Fiscal year:** 2020
- **Award amount:** $225,000
- **Award type:** 1
- **Project period:** 2020-05-20 → 2021-04-30

## Primary source

NIH RePORTER: https://reporter.nih.gov/project-details/10011475

## Citation

> US National Institutes of Health, RePORTER application 10011475, MITOCHONDRIAL RESPIROMETRY IN FROZEN BIOLOGICAL SAMPLES (1R41ES031043-01A1). Retrieved via AI Analytics 2026-05-23 from https://api.ai-analytics.org/grant/nih/10011475. Licensed CC0.

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