# 7-Ketocholesterol Links Sterol Homeostasis to Hedgehog Signaling and Stress–Survival Responses in MSCs from Patients with Acute Myeloid Leukemia

## TL;DR

7-Ketocholesterol affects bone marrow cells in leukemia patients, linking cholesterol balance to survival signals and stress responses.

## Contribution

This study reveals 7-KC as a novel modulator connecting sterol homeostasis to Hedgehog signaling and stress responses in AML MSCs.

## Key findings

- AML-MSCs show a preconditioned state with enhanced Hedgehog and stress–survival responses to 7-KC.
- Healthy donor MSCs are more vulnerable to high 7-KC doses, showing mitochondrial and oxidative stress.
- 7-KC influences lipid transport, redox balance, and mitochondrial dynamics in MSCs.

## Abstract

7-ketocholesterol (7-KC) is a bioactive oxysterol generated under oxidative stress and may contribute to bone marrow niche reprogramming in acute myeloid leukemia (AML), thereby promoting stress tolerance and therapeutic resistance Bone marrow mesenchymal stromal cells (MSCs) from healthy donors and AML patients were exposed to subtoxic 7-KC concentrations for 24 h. We evaluated the ABC transporters involved in lipid transport, multidrug resistance and membrane microdomain remodeling; Hedgehog pathway proteins; stress–survival signaling; redox balance by glutathione measurements, and mitochondrial function and dynamics, including membrane potential and gene expression of mitochondrial fission and fusion regulators. Results were integrated using principal component analysis (PCA), heatmaps, and correlation-based networks. Multivariate analyses revealed an integrated, lineage-dependent response. Healthy donor MSCs showed greater plasticity of the efflux and microdomain axis and higher oxidative and mitochondrial vulnerability at high 7-KC doses. AML-MSCs exhibited a basal preconditioned state phenotype and preferentially routed the response toward Hedgehog and stress–survival modules, accompanied by glutathione expansion and adaptive mitochondrial remodeling. 7-KC acts as a broad modulator of several MSC functions, linking sterol homeostasis to Hedgehog signaling, stress–survival pathways, redox balance, and mitochondrial remodeling, potentially supporting a pro-survival, more therapy-tolerant leukemic niche.