Non-DNA-damaging DNA-PK activation improving hearing and prolonging life due to NAD+ and SIRT upregulation

Honkura, Y.; Suzuki, T.; Kujirai, R.; Kanno, S.; Matsumoto, Y.; Tanaka, Y.; Kowit, H.; Nagatoishi, S.; Tyshkovskiy, A.; Kasahara, T.; Tongu, Y.; Bohan, Z.; Kashiwagi, H.; Saegusa, C.; Fujioka, M.; Usami, R.; Chikuma, S.; Tokifuji, Y.; Matsuhashi, T.; Oikawa, Y.; Komatsu, H.; Murayama, K.; Sugasawa, T.; Nanto-Hara, F.; Taguchi, K.; Saigusa, D.; Suzuki, C.; Sato, T.; Suzuki, J.; Owada, Y.; Niizuma, K.; Endo, H.; Hashimoto, K.; Toyohara, T.; Tsumoto, K.; Anderson, P.; Gladyshev, V. N.; Hayashi, K.-i.; Katori, Y.; Tomioka, Y.; Abe, T.

doi:10.1101/2025.04.18.649305

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bioRxiv · 10.1101/2025.04.18.649305

Non-DNA-damaging DNA-PK activation improving hearing and prolonging life due to NAD+ and SIRT upregulation

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Abstract

Emerging evidence strongly supports a close relationship between age-related hearing loss and frailty, highlighting the importance of early detection and intervention. Recently, we invented a mitochondria-homing drug named mitochonic acid 5 (MA-5), that increases the adenosine triphosphate (ATP) levels, rescue mitochondrial function, and protect tissue damages. Currently, the phase I clinical trial has been finished in Japan (jRCT2031210495) and the phase 2 clinical trial has already been approved by PMDA. Here we show that MA-5 improved various types of hearing loss in mouse models. Structural chemical bioanalysis revealed that MA-5 is a mixture of equal amount of S- and R- enantiomer and both S- and R- enantiomer increase ATP by binding mitochondrial protein, mitofilin. However, S-enantiomer significantly increased the NAD+ levels by binding to the NAD+-producing key enzyme nicotinamide phosphoribosyltransferase (NAMPT). Moreover, the S-enantiomer increased the sirtuin 1 protein by suppressing polyubiquitination induced by tripartite motif containing 28 (TRIM28) phosphorylation which was triggered by DNA-dependent protein kinase (DNA-PK) activation in the absence of DNA damage. Transcriptomic signatures showed that the signature of MA-5 shows an inverse correlation with aging and mortality and is oriented in the same direction as the OSKM-related iPSCs, suggesting the modification of aging pathways. Oral administration of MA-5 to mitochondrial disease model mouse showed increased survival. Our findings suggest that, in addition to enhancing ATP levels, the coordinated regulation of NAD+ metabolism, SIRT protein expression, and DNA-PK activity-constituting a novel therapeutic triad may contribute to the amelioration of hearing impairment and mitochondrial dysfunction, thereby improving life prognosis.

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Honkura, Y., Suzuki, T., Kujirai, R., Kanno, S., Matsumoto, Y., Tanaka, Y., Kowit, H., Nagatoishi, S., Tyshkovskiy, A., Kasahara, T., Tongu, Y., Bohan, Z., Kashiwagi, H., Saegusa, C., Fujioka, M., Usami, R., Chikuma, S., Tokifuji, Y., Matsuhashi, T., Oikawa, Y., Komatsu, H., Murayama, K., Sugasawa, T., Nanto-Hara, F., Taguchi, K., Saigusa, D., Suzuki, C., Sato, T., Suzuki, J., Owada, Y., Niizuma, K., Endo, H., Hashimoto, K., Toyohara, T., Tsumoto, K., Anderson, P., Gladyshev, V. N., Hayashi, K.-i., Katori, Y., Tomioka, Y., Abe, T.. 2025-04-23. Non-DNA-damaging DNA-PK activation improving hearing and prolonging life due to NAD+ and SIRT upregulation. https://doi.org/10.1101/2025.04.18.649305

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