Lactononadecapeptide Upregulates Gene Expression of Neuroplasticity and Cholinergic Activation in Human iPSC-Derived Neurons

Abstract

To clarify the underlying molecular mechanisms of lactononadecapeptide (LNDP), we examined its effects on the gene expression of memory-related neuroplasticity and cholinergic signaling in human induced pluripotent stem cell (hiPSC)-derived neurons, alongside a safety evaluation using PC12 cells. LNDP showed no cytotoxicity and significantly upregulated the expression of genes crucial for neuroplasticity (BDNF, TrkB, NGF) and cholinergic signaling (ChAT, CHRM1, NMDAR NR1) in hiPSC-derived neurons. These findings suggest that LNDP potentially modulates transcriptional pathways related to neural health, supporting its potential value as a functional food ingredient for cognitive decline.

Keywords

Lactononadecapeptide / Human iPSC-derived neurons / Memory / Gene expression

References

[1] Scheltens P, De Strooper B, Kivipelto M, Holstege H, Chételat G, Teunissen CE, et al. Alzheimer’s disease. Lancet 2021, 397, 1577-1590. DOI:10.1016/S0140-6736(20)32205-4

[2] Kowiański P, Lietzau G, Czuba E, Waśkow M, Steliga A, Moryś J. BDNF: A key factor with multipotent impact on brain signaling and synaptic plasticity. Cell Mol. Neurobiol. 2018, 38, 579-593. DOI:10.1007/s10571-017-0510-4

[3] De Sousa Fernandes MS, Ordônio TF, Santos GCJ, Santos LER, Calazans CT, Gomes DA, et al. Effects of physical exercise on neuroplasticity and brain function: a systematic review in human and animal studies. Neural Plast. 2020, 2020, 8856621. DOI:10.1155/2020/8856621

[4] Hozumi S, Nakagawasai O, Tan-No K, Niijima F, Yamadera F, Murata A, et al. Characteristics of changes in cholinergic function and impairment of learning and memory-related behavior induced by olfactory bulbectomy. Behav. Brain Res. 2003, 138, 9-15. DOI:10.1016/S0166-4328(02)00183-3

[5] Riedel G, Platt B, Micheau J. Glutamate receptor function in learning and memory. Behav. Brain Res. 2003, 140, 1-47. DOI:10.1016/S0166-4328(02)00272-3

[6] Hansen KB, Yi F, Perszyk RE, Furukawa H, Wollmuth LP, Gibb AJ, et al. Structure, function, and allosteric modulation of NMDA receptors. J. Gen. Physiol. 2018, 150, 1081-1105. DOI:10.1085/jgp.201812032

[7] Sabbir MG, Speth RC, Albensi BC. Loss of cholinergic receptor muscarinic 1 (CHRM1) protein in the hippocampus and temporal cortex of a subset of individuals with Alzheimer’s disease, Parkinson’s disease, or frontotemporal dementia: Implications for patient survival. J. Alzheimers Dis. 2022, 90, 727-747. DOI:10.3233/JAD-220766

[8] Scott-Solomon E, Kuruvilla R. Mechanisms of neurotrophin trafficking via Trk receptors. Mol. Cell Neurosci. 2018, 91, 25-33. DOI:10.1016/j.mcn.2018.03.013

[9] Tamime AY. Fermented milks: a historical food with modern applications—A review. Eur. J. Clin. Nutr. 2002, 56, S2-S15. DOI:10.1038/sj.ejcn.1601657

[10] Ohsawa K, Uchida N, Ohki K, Nakamura Y, Yokogoshi H. Lactobacillus helveticus-fermented milk improves learning and memory in mice. Nutr. Neurosci. 2015, 18, 232-240. DOI:10.1179/1476830514Y.0000000122

[11] Ohsawa K, Uchida N, Ohki K, Yokogoshi H. Identification of peptides present in sour milk whey that ameliorate scopolamine-induced memory impairment in mice. Int. J. Food Sci. Nutr. 2018, 69, 33-45. DOI:10.1080/09637486.2017.1324564

[12] Ohsawa K, Nakamura F, Uchida N, Mizuno S, Yokogoshi H. Lactobacillus helveticus-fermented milk containing lactononadecapeptide (NIPPLTQTPVVVPPFLQPE) improves cognitive function in healthy middle-aged adults: A randomised, double-blind, placebo-controlled trial. Int. J. Food Sci. Nutr. 2018, 69, 369-376. DOI:10.1080/09637486.2017.1365824

[13] Sasai M, Kato M, Ohsawa K, Sashihara K, Nakamura Y, Kaneko T. Effects of a single dose of tablets containing lactononadecapeptide on cognitive function in healthy adults: A randomized, double-blind, cross-over, placebo-controlled trial. Biosci. Biotechnol. Biochem. 2021, 85, 948-956. DOI:10.1093/bbb/zbaa117

[14] Yamada S, Shirai M, Nakamura F, Ohsawa K, Watanabe M, Koga M. Beneficial effects of food containing lactononadecapeptide on memory function in elderly Japanese subjects-A randomized, double-blind, placebo-controlled study. Food Res. Suppl. 2026, 1, 10006. DOI:10.70322/frs.2026.10006

[15] Benito E, Barco A. CREB’s control of intrinsic and synaptic plasticity: implications for CREB-dependent memory models. Trends Neurosci. 2010, 33, 230-240. DOI:10.1016/j.tins.2010.02.001

[16] Kimura J, Shimizu K, Takito J, Nemoto K, Degawa M, Yokosuka A, et al. Nobiletin reduces intracellular and extracellular β-amyloid in iPS cell-derived Alzheimer’s disease model neurons. Biol. Pharm. Bull. 2018, 41, 451-457. DOI:10.1248/bpb.b17-00364

[17] Kimura J, Shimizu K, Takito J, Nemoto K, Degawa M, Yokosuka A, et al. Upregulatory effects of nobiletin, a citrus flavonoid with anti-dementia activity, on the gene expression of mAChR, ChAT, and CBP. Planta Med. Lett. 2015, 2, e12-e14. DOI:10.1055/s-0035-1545937

[18] Livak KJ, Schmittgen TD. Analysis of relative gene expression data using real-time quantitative PCR and the 2^-ΔΔC_T method. Methods 2001, 25, 402-408. DOI:10.1006/meth.2001.1262

[19] Nakatani E, Sasai M, Miyazaki H, Tanaka S, Hirota T, Okura T. Investigating the transepithelial transport and enzymatic stability of lactononadecapeptide (NIPPLTQTPVVVPPFLQPE), a 19-amino acid casein-derived peptide in Caco-2 cells. J. Agric. Food Chem. 2024, 72, 12719-12724. DOI:10.1021/acs.jafc.4c00974