AAV administration drives a nephrotoxic response through NFκB in kidney organoids. a Left: Reside epifluorescent microscopy informing AAV dose and length (AAV2 depicted). Proper: Multiplexed immunostaining and quantification of GFP reporter supply in LTL+ proximal tubules, CDH1+ distal tubules, PDGFR-β+ interstitial cells, CD31+ endothelia, and PODXL+ podocytes utilizing variable of AAV serotypes. Scale bar 200 μm. b Left: Immunostaining and quantification of AAV2-based supply of Cas9 and mCherry within the nephron epithelial cells of CRISPR samples. Scale bar 50 μm. ***p Sign Transduction and Focused Remedy (2025). DOI: 10.1038/s41392-025-02336-2
Ryuji Morizane, MD, Ph.D., of the Division of Medication at Massachusetts Basic Hospital, is the senior/corresponding writer of a brand new paper printed in Sign Transduction and Focused Remedy, titled “AAV for gene therapy drives a nephrotoxic response via NFκB in kidney organoids.”
On this interview, Dr. Morizane discusses the research.
How would you summarize your research for a lay viewers?
Gene remedy holds nice promise for treating severe genetic illnesses equivalent to Duchenne muscular dystrophy (DMD). Nonetheless, surprising poisonous negative effects, together with affected person deaths in some DMD trials, have raised main security issues.
These dangers are sometimes missed in animal research, revealing a essential flaw in present preclinical testing strategies. Actually, fewer than 15% of medicine that enter scientific trials ever obtain FDA approval, largely as a result of conventional lab fashions fail to foretell how the human physique will reply to the remedy.
To handle this vital hole, our research used human stem cell-derived kidney organoids—lab-grown mini kidneys—to check the security of gene modifying delivered by adeno-associated virus (AAV), a typical instrument in scientific trials.
We discovered that AAV2 (one among a number of variants of the AAV virus at the moment used to ship gene remedy remedy) precipitated vital hurt to kidney cells by triggering irritation, DNA harm, and fibrosis. This harm occurred by the NFκB pathway, even with none gene modifying happening. Encouragingly, an current drug that blocks this pathway was in a position to forestall the harm from occurring within the organoids with out interfering with the mechanism of gene supply.
These outcomes present that human-centric lab fashions equivalent to organoids are vitally wanted to detect hidden dangers and enhance the security of gene therapies earlier than they attain sufferers.
What query had been you investigating?
We investigated whether or not human stem cell-derived kidney organoids, an rising and more and more acknowledged know-how, can function a extra correct preclinical mannequin for evaluating the therapeutic efficacy and potential negative effects of AAVs utilized in gene remedy.
Which strategies or strategy did you utilize?
We used human stem cell-derived kidney organoids, lab-grown mini-kidneys initially developed by our group at Mass Basic Brigham, as a preclinical platform to guage the security and efficacy of AAV-based gene remedy.
What did you discover?
We discovered that the AAV2 variant induced vital toxicity in kidney organoids—triggering irritation, DNA harm, fibrosis, and mobile senescence, significantly in proximal tubules. These results, which occurred even with out gene modifying, had been pushed by activation of the NFκB signaling pathway.
Importantly, remedy with bardoxolone methyl considerably lowered these dangerous responses with out impairing AAV-mediated gene supply. This means that AAV toxicity, not gene modifying, is a main contributor to noticed tissue harm.
Extra broadly, our research reveals that these human-specific negative effects, together with potential preventive methods, can now be evaluated utilizing human kidney organoids.
This represents a robust addition to the drug growth pipeline, serving to to establish hidden dangers earlier and complementing present preclinical testing to make gene therapies safer and more practical earlier than reaching sufferers.
What are the implications?
Broader recognition and regulatory acceptance of organoids are vitally wanted to enhance affected person security, cut back trial failures, and in the end speed up the event of more practical and customized therapies.
Importantly, organoid-based approaches usually are not meant to interchange animal fashions totally, however to enrich them, including a layer of human-specific perception that present animal testing usually can not present.
What are the following steps?
Future work will deal with bettering the physiological relevance of kidney organoids by incorporating vascular constructions and extra mature cell sorts to raised replicate in vivo kidney operate. We additionally purpose to increase this organoid-based security screening strategy to different organ programs and AAV serotypes, broadening its utility in gene remedy growth.
To make sure reproducibility and regulatory applicability, it can even be important to deal with batch-to-batch variation in organoid differentiation. Growing standardized protocols for producing kidney organoids and for assessing gene therapeutic merchandise inside these fashions will likely be essential for his or her integration into preclinical pipelines.
Extra info:
Navin Gupta et al, AAV for gene remedy drives a nephrotoxic response through NFκB in kidney organoids, Sign Transduction and Focused Remedy (2025). DOI: 10.1038/s41392-025-02336-2
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Massachusetts Basic Hospital
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Q&A: How organoids may make gene remedy trials safer by figuring out hidden dangers early on (2025, August 19)
retrieved 19 August 2025
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