{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,6,3]],"date-time":"2026-06-03T02:01:55Z","timestamp":1780452115489,"version":"3.54.1"},"reference-count":26,"publisher":"MDPI AG","issue":"4","license":[{"start":{"date-parts":[[2025,2,18]],"date-time":"2025-02-18T00:00:00Z","timestamp":1739836800000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"name":"USA Health Mitchell Cancer Institute"},{"name":"Norma Livingston Ovarian Cancer Foundation"}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["IJMS"],"abstract":"<jats:p>Dihydronicotinamide rioside (NRH), the reduced form of nicotinamide riboside (NR), is a recently identified, naturally occurring precursor of arguably the most crucial cofactor for cellular function, nicotinamide adenine dinucleotide (NAD+). Recent investigation suggests that NRH is more adept at increasing NAD+ stores than traditional NAD+ precursors, and such extreme NAD+ boosting via NRH supplementation induces cytotoxicity in certain cellular contexts. It has also been shown that the lack of functional BRCA protein in epithelial ovarian cancer (EOC) directly impacts intracellular NAD+ levels. Given that altered cellular metabolism and DNA repair mechanisms are central alterations in EOC, and these processes are functionally dependent on NAD+, we sought to assess whether NRH supplementation in EOC cell lines enhanced cellular cytotoxicity alone and in combination with standard therapeutic agents. Significant cytotoxicity was noted in NRH treated cells (~40%) with minimal cell death in the nicotinic acid (NA)-treated lines. Levels of NAD(P)H were confirmed to have increased with NRH supplementation, albeit at different levels among the different cell lines. Overall, the cytotoxicity associated with NRH supplementation appears to be independent of ROS generation. Strikingly, NRH supplementation enhanced cytotoxicity of carboplatin in OVCAR8, but not ES2 or SKOV3. Paclitaxel cytotoxicity was also enhanced by the addition of NRH in OVCAR8, but not ES2 or SKOV3 cell lines. NA supplementation had no effect on baseline treatment-induced cytotoxicity. PARP inhibition by olaparib requires NAD+. Interestingly, NRH supplementation enhanced olaparib cytotoxicity in SKOV3 and OVCAR8, but not ES2 cells. NRH in combination with olaparib completely altered mitochondrial respiration, thereby shutting down energy consumption, which would lead to cell death. Coupled together with expression data of key enzymes required for NRH\/NAD metabolism, this could be key in understanding mechanisms of cell death with NRH supplementation. Here, we showed that in the context of EOC, exploitation of the NAD+ bioenergetic phenotype through NRH supplementation is a biologically feasible strategy to enhance the response of traditional therapy with potentially minimal toxicity. These data suggest several potential mechanisms by which cellular NAD+ availability impacts treatment efficacy and resistance and highlights the potential utility of NAD+ metabolomics as a biomarker to guide treatment decisions.<\/jats:p>","DOI":"10.3390\/ijms26041719","type":"journal-article","created":{"date-parts":[[2025,2,18]],"date-time":"2025-02-18T03:47:06Z","timestamp":1739850426000},"page":"1719","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":2,"title":["NAD+ Boosting Through NRH Supplementation Enhances Treatment Efficacy in EOC In Vitro"],"prefix":"10.3390","volume":"26","author":[{"ORCID":"https:\/\/orcid.org\/0000-0003-3233-5537","authenticated-orcid":false,"given":"Kevin J.","family":"Lee","sequence":"first","affiliation":[{"name":"USA Health Mitchell Cancer Institute, University of South Alabama, Mobile, AL 36604, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Sagar","family":"Chokshi","sequence":"additional","affiliation":[{"name":"USA Health Mitchell Cancer Institute, University of South Alabama, Mobile, AL 36604, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Tanvi","family":"Joshi","sequence":"additional","affiliation":[{"name":"USA Health Mitchell Cancer Institute, University of South Alabama, Mobile, AL 36604, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-1560-6406","authenticated-orcid":false,"given":"Mackenzie","family":"Cummings","sequence":"additional","affiliation":[{"name":"USA Health Mitchell Cancer Institute, University of South Alabama, Mobile, AL 36604, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0009-0004-3202-9294","authenticated-orcid":false,"given":"Catherine E.","family":"Lyons","sequence":"additional","affiliation":[{"name":"USA Health Mitchell Cancer Institute, University of South Alabama, Mobile, AL 36604, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Mary Howard","family":"Singleton","sequence":"additional","affiliation":[{"name":"USA Health Mitchell Cancer Institute, University of South Alabama, Mobile, AL 36604, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Elizabeth","family":"Catranis","sequence":"additional","affiliation":[{"name":"USA Health Mitchell Cancer Institute, University of South Alabama, Mobile, AL 36604, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Luciana","family":"Madiera da Silva","sequence":"additional","affiliation":[{"name":"USA Health Mitchell Cancer Institute, University of South Alabama, Mobile, AL 36604, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-5493-0199","authenticated-orcid":false,"given":"Faisal","family":"Hayat","sequence":"additional","affiliation":[{"name":"Department of Pharmacology, University of South Alabama, Mobile, AL 36688, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-9626-2405","authenticated-orcid":false,"given":"Marie","family":"Migaud","sequence":"additional","affiliation":[{"name":"Department of Pharmacology, University of South Alabama, Mobile, AL 36688, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Jennifer","family":"Scalici","sequence":"additional","affiliation":[{"name":"Winship Cancer Institute, Emory University School of Medicine, Atlanta, GA 30322, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1968","published-online":{"date-parts":[[2025,2,18]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"31","DOI":"10.1158\/2159-8290.CD-21-1059","article-title":"Hallmarks of Cancer: New Dimensions","volume":"12","author":"Hanahan","year":"2022","journal-title":"Cancer Discov."},{"key":"ref_2","doi-asserted-by":"crossref","first-page":"101195","DOI":"10.1016\/j.molmet.2021.101195","article-title":"Role of NAD+ in regulating cellular and metabolic signaling pathways","volume":"49","author":"Amjad","year":"2021","journal-title":"Mol. Metab."},{"key":"ref_3","doi-asserted-by":"crossref","first-page":"822","DOI":"10.1038\/s41580-024-00752-w","article-title":"Regulation of and challenges in targeting NAD+ metabolism","volume":"25","author":"Migaud","year":"2024","journal-title":"Nat. Rev. Mol. Cell Biol."},{"key":"ref_4","doi-asserted-by":"crossref","first-page":"2","DOI":"10.1038\/s41392-020-00354-w","article-title":"NAD+ metabolism, stemness, the immune response, and cancer","volume":"6","author":"Navas","year":"2021","journal-title":"Sig. Transduct. Target. Ther."},{"key":"ref_5","doi-asserted-by":"crossref","unstructured":"Yong, J., Cai, S., and Zeng, Z. (2023). Targeting NAD+ metabolism: Dual roles in cancer treatment. Front. Immunol., 14.","DOI":"10.3389\/fimmu.2023.1269896"},{"key":"ref_6","doi-asserted-by":"crossref","unstructured":"Morandi, F., Horenstein, A.L., and Malavasi, F. (2021). The Key Role of NAD+ in Anti-Tumor Immune Response: An Update. Front. Immunol., 12.","DOI":"10.3389\/fimmu.2021.658263"},{"key":"ref_7","doi-asserted-by":"crossref","first-page":"274","DOI":"10.1016\/j.phrs.2016.10.027","article-title":"NAD+ salvage pathway in cancer metabolism and therapy","volume":"114","author":"Kennedy","year":"2016","journal-title":"Pharmacol. Res."},{"key":"ref_8","doi-asserted-by":"crossref","first-page":"2584","DOI":"10.1091\/mbc.E18-10-0650","article-title":"NAD+ consumption by PARP1 in response to DNA damage triggers metabolic shift critical for damaged cell survival","volume":"30","author":"Murata","year":"2019","journal-title":"Mol. Biol. Cell"},{"key":"ref_9","doi-asserted-by":"crossref","first-page":"610","DOI":"10.1038\/nrm.2017.53","article-title":"The multifaceted roles of PARP1 in DNA repair and chromatin remodelling","volume":"18","author":"Nussenzweig","year":"2017","journal-title":"Nat. Rev. Mol. Cell Biol."},{"key":"ref_10","doi-asserted-by":"crossref","unstructured":"Rose, M., Burgess, J.T., O\u2019Byrne, K., Richard, D.J., and Bolderson, E. (2020). PARP Inhibitors: Clinical Relevance, Mechanisms of Action and Tumor Resistance. Front. Cell Dev. Biol., 8.","DOI":"10.3389\/fcell.2020.564601"},{"key":"ref_11","doi-asserted-by":"crossref","first-page":"2495","DOI":"10.1056\/NEJMoa1810858","article-title":"Maintenance Olaparib in Patients with Newly Diagnosed Advanced Ovarian Cancer","volume":"379","author":"Moore","year":"2018","journal-title":"N. Engl. J. Med."},{"key":"ref_12","doi-asserted-by":"crossref","first-page":"2416","DOI":"10.1056\/NEJMoa1911361","article-title":"Olaparib plus Bevacizumab as First-Line Maintenance in Ovarian Cancer","volume":"381","author":"Pautier","year":"2019","journal-title":"N. Engl. J. Med."},{"key":"ref_13","doi-asserted-by":"crossref","first-page":"12","DOI":"10.3322\/caac.21820","article-title":"Cancer statistics, 2024","volume":"74","author":"Siegel","year":"2024","journal-title":"CA Cancer J. Clinicians."},{"key":"ref_14","doi-asserted-by":"crossref","first-page":"2391","DOI":"10.1056\/NEJMoa1910962","article-title":"Niraparib in Patients with Newly Diagnosed Advanced Ovarian Cancer","volume":"381","author":"Pothuri","year":"2019","journal-title":"N. Engl. J. Med."},{"key":"ref_15","doi-asserted-by":"crossref","first-page":"604","DOI":"10.1021\/acschembio.0c00757","article-title":"Chemical and Biochemical Reactivity of the Reduced Forms of Nicotinamide Riboside","volume":"16","author":"Makarov","year":"2021","journal-title":"ACS Chem. Biol."},{"key":"ref_16","doi-asserted-by":"crossref","unstructured":"Sonavane, M., Hayat, F., Makarov, M., Migaud, M.E., and Gassman, N.R. (2020). Dihydronicotinamide riboside promotes cell-specific cytotoxicity by tipping the balance between metabolic regulation and oxidative stress. PLoS ONE, 15.","DOI":"10.1371\/journal.pone.0242174"},{"key":"ref_17","doi-asserted-by":"crossref","first-page":"9295","DOI":"10.1074\/jbc.RA118.005772","article-title":"Dihydronicotinamide riboside is a potent NAD+ concentration enhancer in vitro and in vivo","volume":"294","author":"Yang","year":"2019","journal-title":"J. Biol. Chem."},{"key":"ref_18","doi-asserted-by":"crossref","first-page":"693","DOI":"10.1038\/s41467-019-08530-5","article-title":"NADP+ is an endogenous PARP inhibitor in DNA damage response and tumor suppression","volume":"10","author":"Bian","year":"2019","journal-title":"Nat. Commun."},{"key":"ref_19","doi-asserted-by":"crossref","unstructured":"Chini, C.C.S., Peclat, T.R., Gomez, L.S., Zeidler, J.D., Warner, G.M., Kashyap, S., Mazdeh, D.Z., Hayat, F., Migaud, M.E., and Paulus, A. (2022). Dihydronicotinamide Riboside Is a Potent NAD+ Precursor Promoting a Pro-Inflammatory Phenotype in Macrophages. Front. Immunol., 13.","DOI":"10.3389\/fimmu.2022.840246"},{"key":"ref_20","doi-asserted-by":"crossref","unstructured":"Ciarlo, E., Joffraud, M., Hayat, F., Giner, M.P., Giroud-Gerbetant, J., Sanchez-Garcia, J.L., Rumpler, M., Moco, S., Migaud, M.E., and Cant\u00f3, C. (2022). Nicotinamide Riboside and Dihydronicotinic Acid Riboside Synergistically Increase Intracellular NAD+ by Generating Dihydronicotinamide Riboside. Nutrients, 14.","DOI":"10.3390\/nu14132752"},{"key":"ref_21","doi-asserted-by":"crossref","first-page":"1286","DOI":"10.1038\/s41467-018-03421-7","article-title":"Chronic nicotinamide riboside supplementation is well-tolerated and elevates NAD+ in healthy middle-aged and older adults","volume":"9","author":"Martens","year":"2018","journal-title":"Nat. Commun."},{"key":"ref_22","doi-asserted-by":"crossref","first-page":"6206","DOI":"10.1158\/1078-0432.CCR-11-0736","article-title":"Enhancement of carboplatin-mediated lung cancer cell killing by simultaneous disruption of glutathione and thioredoxin metabolism","volume":"17","author":"Fath","year":"2011","journal-title":"Clin. Cancer Res."},{"key":"ref_23","doi-asserted-by":"crossref","first-page":"231","DOI":"10.1038\/s41392-020-00326-0","article-title":"NADPH homeostasis in cancer: Functions, mechanisms and therapeutic implications","volume":"5","author":"Ju","year":"2020","journal-title":"Sig. Transduct. Target. Ther."},{"key":"ref_24","doi-asserted-by":"crossref","first-page":"267","DOI":"10.1038\/s41419-022-04713-z","article-title":"Complex roles of nicotinamide N-methyltransferase in cancer progression","volume":"13","author":"Wang","year":"2022","journal-title":"Cell Death Dis."},{"key":"ref_25","unstructured":"Redpath, P., Cunningham, R., Crossey, K., Nygaard, R., Erickson, A., Miguad, M.E., and Storjohann, A. (2021). Efficient and Scalable Syntheses of Nicotinoyl Ribosides and Reduced Nicotinoyl Ribosides, Modified Derivatives Thereof, Phosphorylated Analogs Thereof, Adenylyl Dinucleotide Conjugates Thereof, and Novel Crystalline Forms Thereof. (US-20210347806-A1), Unified Patents, Available online: https:\/\/portal.unifiedpatents.com\/patents\/patent\/US-20210347806-A1."},{"key":"ref_26","doi-asserted-by":"crossref","unstructured":"Hernandez, A., Sonavane, M., Smith, K.R., Seiger, J., Migaud, M.E., and Gassman, N.R. (2022). Dihydroxyacetone suppresses mTOR nutrient signaling and induces mitochondrial stress in liver cells. PLoS ONE, 17.","DOI":"10.1371\/journal.pone.0278516"}],"container-title":["International Journal of Molecular Sciences"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.mdpi.com\/1422-0067\/26\/4\/1719\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,10,9]],"date-time":"2025-10-09T16:36:30Z","timestamp":1760027790000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.mdpi.com\/1422-0067\/26\/4\/1719"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2025,2,18]]},"references-count":26,"journal-issue":{"issue":"4","published-online":{"date-parts":[[2025,2]]}},"alternative-id":["ijms26041719"],"URL":"https:\/\/doi.org\/10.3390\/ijms26041719","relation":{},"ISSN":["1422-0067"],"issn-type":[{"value":"1422-0067","type":"electronic"}],"subject":[],"published":{"date-parts":[[2025,2,18]]}}}