Abstract
Post-stroke cognitive impairment (PSCI) is a clinical outcome in around 30% of post-stroke survivors. BDNF is a major gene in this regard. It is regulated by circadian rhythm. The circadian genes are correlated with stroke timings at molecular level. However, studies suggesting the role of these on susceptibility to PSCI are limited. We aim here to determine: (a) genetic risk variants in circadian clock genes, BDNF and (b) dysregulation in expression level of CLOCK, BMAL1, and BDNF that may be associated with PSCI. BDNF (rs6265G/A, rs56164415C/T), CLOCK (rs1801260T/C, rs4580704G/C), and CRY2 (rs2292912C/G) genes variants were genotyped among 119 post-stroke survivors and 292 controls from Eastern part of India. In addition, we analyzed their gene expression in Peripheral blood Mononuclear cells (PBMC) from 15 PSCI cases and 12 controls. The mRNA data for BDNF was further validated by its plasma level through ELISA (n = 38). Among the studied variants, only rs4580704/CLOCK showed an overall association with PSCI (P = 0.001) and lower Bengali Mini-Mental State Examination (BMSE) score. Its ‘C’ allele showed a correlation with attention deficiency. The language and memory impairments showed association with rs6265/BDNF, while the ‘CC’ genotype of rs2292912/CRY2 negatively influenced language and executive function. A significant decrease in gene expression for CLOCK and BDNF in PBMC (influenced by specific genotypes) of PSCI patients was observed than controls. Unlike Pro-BDNF, plasma-level mBDNF was also lower in them. Our results suggest the genetic variants in CLOCK, CRY2, and BDNF as risk factors for PSCI among eastern Indians. At the same time, a lowering expression of CLOCK and BDNF genes in PSCI patients than controls describes their transcriptional dysregulation as underlying mechanism for post-stroke cognitive decline.
Similar content being viewed by others
Data availability
The data that support the findings of this study are available from the corresponding author upon request.
References
Al-Qazzaz, N. K., Ali, S. H., Ahmad, S. A., Islam, S., & Mohamad, K. (2014). Cognitive impairment and memory dysfunction after a stroke diagnosis: A post-stroke memory assessment. Neuropsychiatric Disease and Treatment, 10, 1677–1691. https://doi.org/10.2147/NDT.S67184
Balkaya, M., & Cho, S. (2019). Genetics of stroke recovery: BDNF val66met polymorphism in stroke recovery and its interaction with aging. Neurobiology of Disease, 126, 36–46. https://doi.org/10.1016/j.nbd.2018.08.009
Bao, M. H., Zhu, S. Z., Gao, X. Z., Sun, H. S., & Feng, Z. P. (2018). Meta-analysis on the association between brain-derived neurotrophic factor polymorphism rs6265 and ischemic stroke, poststroke depression. Journal of Stroke and Cerebrovascular Diseases : The Official Journal of National Stroke Association, 27(6), 1599–1608. https://doi.org/10.1016/j.jstrokecerebrovasdis.2018.01.010
Beker, M. C., Caglayan, B., Yalcin, E., Caglayan, A. B., Turkseven, S., Gurel, B., Kelestemur, T., Sertel, E., Sahin, Z., Kutlu, S., Kilic, U., Baykal, A. T., & Kilic, E. (2018). Time-of-day dependent neuronal injury after ischemic stroke: implication of circadian clock transcriptional factor Bmal1 and survival kinase AKT. Molecular Neurobiology, 55(3), 2565–2576. https://doi.org/10.1007/s12035-017-0524-4
Biswas, A., Sadhukhan, D., Biswas, A., Das, S. K., Banerjee, T. K., Bal, P. S., Pal, S., Ghosh, A., Ray, K., & Ray, J. (2021). Identification of GBA mutations among neurodegenerative disease patients from eastern India. Neuroscience Letters, 751, 135816. https://doi.org/10.1016/j.neulet.2021.135816
Carter, B., Justin, H. S., Gulick, D., & Gamsby, J. J. (2021). The molecular clock and neurodegenerative disease: A stressful time. Frontiers in Molecular Biosciences, 8, 644747. https://doi.org/10.3389/fmolb.2021.644747
Chen, Z. Y., Patel, P. D., Sant, G., Meng, C. X., Teng, K. K., Hempstead, B. L., & Lee, F. S. (2004). Variant brain-derived neurotrophic factor (BDNF) (Met66) alters the intracellular trafficking and activity-dependent secretion of wild-type BDNF in neurosecretory cells and cortical neurons. The Journal of Neuroscience : The Official Journal of the Society for Neuroscience, 24(18), 4401–4411. https://doi.org/10.1523/JNEUROSCI.0348-04.2004
Cullen, B., O’Neill, B., Evans, J. J., Coen, R. F., & Lawlor, B. A. (2007). A review of screening tests for cognitive impairment. Journal of Neurology, Neurosurgery, and Psychiatry, 78(8), 790–799. https://doi.org/10.1136/jnnp.2006.095414
D’Agostino, Y., Frigato, E., Noviello, T. M. R., Toni, M., Frabetti, F., Cigliano, L., Ceccarelli, M., Sordino, P., Cerulo, L., Bertolucci, C., & D’Aniello, S. (2022). Loss of circadian rhythmicity in bdnf knockout zebrafish larvae. iScience, 25(4), 104054. https://doi.org/10.1016/j.isci.2022.104054
Das, G., Dubey, S., Sinharoy, U., Mukherjee, A., Banerjee, S., Lahiri, D., & Biswas, A. (2021). Clinical and radiological profile of posterior cortical atrophy and comparison with a group of typical Alzheimer disease and amnestic mild cognitive impairment. Acta Neurologica Belgica, 121(4), 1009–1018. https://doi.org/10.1007/s13760-020-01547-4
Di Lazzaro, V., Pellegrino, G., Di Pino, G., Corbetto, M., Ranieri, F., Brunelli, N., Paolucci, M., Bucossi, S., Ventriglia, M. C., Brown, P., & Capone, F. (2015). Val66Met BDNF gene polymorphism influences human motor cortex plasticity in acute stroke. Brain Stimulation, 8(1), 92–96. https://doi.org/10.1016/j.brs.2014.08.006
Eckel-Mahan, K. L., & Storm, D. R. (2009). Circadian rhythms and memory: Not so simple as cogs and gears. EMBO Reports, 10(6), 584–591. https://doi.org/10.1038/embor.2009.123
Han, Z., Qi, L., Xu, Q., Xu, M., Cai, L., Wong, J., Hu, X., Luo, X., Wang, J., Zhang, Y., Li, Y., & Wang, Q. M. (2020). BDNF met allele is associated with lower cognitive function in poststroke rehabilitation. Neurorehabilitation and Neural Repair, 34(3), 247–259. https://doi.org/10.1177/1545968320902127
Hassan, T. M., & Yarube, I. U. (2018). Peripheral brain-derived neurotrophic factor is reduced in stroke survivors with cognitive impairment. Pathophysiology : The Official Journal of the International Society for Pathophysiology, 25(4), 405–410. https://doi.org/10.1016/j.pathophys.2018.08.003
Indian Genome Variation Consortium. (2005). The Indian Genome Variation database (IGVdb): A project overview. Human Genetics, 118(1), 1–11. https://doi.org/10.1007/s00439-005-0009-9
Johns, M. B., Jr., & Paulus-Thomas, J. E. (1989). Purification of human genomic DNA from whole blood using sodium perchlorate in place of phenol. Analytical Biochemistry, 180(2), 276–278. https://doi.org/10.1016/0003-2697(89)90430-2
Karantali, E., Kazis, D., Papavasileiou, V., Prevezianou, A., Chatzikonstantinou, S., Petridis, F., McKenna, J., Luca, A. C., Trus, C., Ciobica, A., & Mavroudis, I. (2021). Serum BDNF levels in acute stroke: A systematic review and meta-analysis. Medicina (kaunas, Lithuania), 57(3), 297. https://doi.org/10.3390/medicina57030297
Kim, J. M., Stewart, R., Park, M. S., Kang, H. J., Kim, S. W., Shin, I. S., Kim, H. R., Shin, M. G., Cho, K. H., & Yoon, J. S. (2012). Associations of BDNF genotype and promoter methylation with acute and long-term stroke outcomes in an East Asian cohort. PLoS ONE, 7(12), e51280. https://doi.org/10.1371/journal.pone.0051280
Levine, D. A., Wadley, V. G., Langa, K. M., Unverzagt, F. W., Kabeto, M. U., Giordani, B., Howard, G., Howard, V. J., Cushman, M., Judd, S. E., & Galecki, A. T. (2018). Risk factors for poststroke cognitive decline: The REGARDS study (reasons for geographic and racial differences in stroke). Stroke, 49(4), 987–994. https://doi.org/10.1161/STROKEAHA.117.018529
Liang, F. Q., Walline, R., & Earnest, D. J. (1998). Circadian rhythm of brain-derived neurotrophic factor in the rat suprachiasmatic nucleus. Neuroscience Letters, 242(2), 89–92. https://doi.org/10.1016/s0304-3940(98)00062-7
Mirowska-Guzel, D., Gromadzka, G., Czlonkowski, A., & Czlonkowska, A. (2012). BDNF -270 C>T polymorphisms might be associated with stroke type and BDNF -196 G>A corresponds to early neurological deficit in hemorrhagic stroke. Journal of Neuroimmunology, 249(1–2), 71–75. https://doi.org/10.1016/j.jneuroim.2012.04.011
Monti, P., Iodice, S., Tarantini, L., Sacchi, F., Ferrari, L., Ruscica, M., Buoli, M., Vigna, L., Pesatori, A. C., & Bollati, V. (2021). Effects of PM exposure on the methylation of clock genes in a population of subjects with overweight or obesity. International Journal of Environmental Research and Public Health, 18(3), 1122. https://doi.org/10.3390/ijerph18031122
Moore, D. F., Li, H., Jeffries, N., Wright, V., Cooper, R. A., Jr., Elkahloun, A., Gelderman, M. P., Zudaire, E., Blevins, G., Yu, H., Goldin, E., & Baird, A. E. (2005). Using peripheral blood mononuclear cells to determine a gene expression profile of acute ischemic stroke: A pilot investigation. Circulation, 111(2), 212–221. https://doi.org/10.1161/01.CIR.0000152105.79665.C6
Niu, Y., Wan, C., Zhou, B., Wang, J., Wang, J., Chen, X., Li, R., Wang, X., Liu, W., & Wang, Y. (2018). Aerobic exercise relieved vascular cognitive impairment via NF-κB/miR-503/BDNF pathway. American Journal of Translational Research, 10(3), 753–761.
Notaras, M., & van den Buuse, M. (2019). Brain-derived neurotrophic factor (BDNF): Novel insights into regulation and genetic variation. The Neuroscientist : A Review Journal Bringing Neurobiology, Neurology and Psychiatry, 25(5), 434–454. https://doi.org/10.1177/1073858418810142
Ozburn, A. R., Purohit, K., Parekh, P. K., Kaplan, G. N., Falcon, E., Mukherjee, S., Cates, H. M., & McClung, C. A. (2016). Functional implications of the CLOCK 3111T/C single-nucleotide polymorphism. Frontiers in Psychiatry, 7, 67. https://doi.org/10.3389/fpsyt.2016.00067
Pramong, R., Govitrapong, P., & Phansuwan-Pujito, P. (2017). Relationship between circadian clock genes and the neurotrophic factor genes in rat hippocampus. Journal of the Medical Association of Thailand, 100(10), 32.
Rezaei, S., Asgari-Mobarake, K., Keshavarz, P., Tolami, H. F., Saravani, M. F., Saberi, A., Hosseininezhad, M., Bakhshayesh- Eghbali, B., & Kouchakinejad-Eramsadati, L. (2017). Brain-derived neurotrophic factor (BDNF) Val66met (rs6265) polymorphism associated with global and multi-domain cognitive impairment in ischemic stroke patients. Activitas Nervosa Superior, 59, 28–36. https://doi.org/10.1007/s41470-017-0001-4
Srithumsuk, W., Kabayama, M., Gondo, Y., Masui, Y., Akagi, Y., Klinpudtan, N., Kiyoshige, E., Godai, K., Sugimoto, K., Akasaka, H., Takami, Y., Takeya, Y., Yamamoto, K., Ikebe, K., Ogawa, M., Inagaki, H., Ishizaki, T., Arai, Y., Rakugi, H., & Kamide, K. (2020). The importance of stroke as a risk factor of cognitive decline in community dwelling older and oldest peoples: The SONIC study. BMC Geriatrics, 20(1), 24. https://doi.org/10.1186/s12877-020-1423-5
Sadhukhan D., Biswas A., Biswas a., Ray K., & Ray, J. (2022). Genetic polymorphisms in DRD4 and risk for Parkinson’s disease among eastern Indians. Neurology India, 70(2), 729–732. https://doi.org/10.4103/0028-3886.344670
Stanne, T. M., Tjärnlund-Wolf, A., Olsson, S., Jood, K., Blomstrand, C., & Jern, C. (2014). Genetic variation at the BDNF locus: Evidence for association with long-term outcome after ischemic stroke. PLoS ONE, 9(12), e114156. https://doi.org/10.1371/journal.pone.0114156
Tischkau, S. A., Cohen, J. A., Stark, J. T., Gross, D. R., & Bottum, K. M. (2007). Time-of-day affects expression of hippocampal markers for ischemic damage induced by global ischemia. Experimental Neurology, 208(2), 314–322. https://doi.org/10.1016/j.expneurol.2007.09.003
Valenzuela, F. J., Vera, J., Venegas, C., Muñoz, S., Oyarce, S., Muñoz, K., & Lagunas, C. (2016). Evidences of polymorphism associated with circadian system and risk of pathologies: A review of the literature. International Journal of Endocrinology, 2016, 2746909. https://doi.org/10.1155/2016/2746909
Wiebking, N., Maronde, E., & Rami, A. (2013). Increased neuronal injury in clock gene Per-1 deficient-mice after cerebral ischemia. Current Neurovascular Research, 10(2), 112–125. https://doi.org/10.2174/1567202611310020004
Zhao, J., Wu, H., Zheng, L., Weng, Y., & Mo, Y. (2013). Brain-derived neurotrophic factor G196A polymorphism predicts 90-day outcome of ischemic stroke in Chinese: A novel finding. Brain Research, 1537, 312–318. https://doi.org/10.1016/j.brainres.2013.08.061
Zheng, F., Zhou, X., Moon, C., & Wang, H. (2012). Regulation of brain-derived neurotrophic factor expression in neurons. International Journal of Physiology, Pathophysiology and Pharmacology, 4(4), 188–200.
Zsuga, J., More, C. E., Erdei, T., Papp, C., Harsanyi, S., & Gesztelyi, R. (2018). Blind spot for sedentarism: redefining the diseasome of physical inactivity in view of circadian system and the Irisin/BDNF axis. Frontiers in Neurology, 9, 818. https://doi.org/10.3389/fneur.2018.00818
Acknowledgements
The authors are thankful to the patients, family members, and healthy individuals who participated in the study.
Funding
The study has been supported by Post-Doctoral fellowship grants from the Department of Science & Technology, Govt. of India, under Cognitive Science Research Initiative Program to first author DS [DST/CSRI-PDF/2021/12 & DST/CSRI/PDF-21/2018] and the second author AB [DST/CSRI-P/2017/22].
Author information
Authors and Affiliations
Contributions
DS, ID, and AB were responsible for the concept, study design, experimental work, and manuscript preparation. SM, PM, PM, and GP had put effort in data analysis (both clinical and genetic) and manuscript preparation. KC did the statistical analysis. AB, BKR, and TKB evaluated patients and blood collections as being clinical collaborates. While, SPH provided the instrumentation facility in S. N. Pradhan Center for Neurosciences, University of Calcutta and provide his input for manuscript preparation. All authors read the draft, provided their inputs, and agreed on the final version of the manuscript.
Corresponding authors
Ethics declarations
Conflict of interest
The authors declare that they have no conflicts of interest.
Ethical approval
All procedures performed in studies involving human participants were in accordance with the ethical standards of the institutional and national research committee as well as the 1964 Helsinki Declaration and its later amendments.
Informed consent
Informed consent from all the participants were received prior to clinical data and sample collection.
Additional information
Publisher's Note
Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.
Rights and permissions
Springer Nature or its licensor (e.g. a society or other partner) holds exclusive rights to this article under a publishing agreement with the author(s) or other rightsholder(s); author self-archiving of the accepted manuscript version of this article is solely governed by the terms of such publishing agreement and applicable law.
About this article
Cite this article
Sadhukhan, D., Biswas, A., Mishra, S. et al. Genetic Variations and Altered Blood mRNA Level of Circadian Genes and BDNF as Risk Factors of Post-Stroke Cognitive Impairment Among Eastern Indians. Neuromol Med 25, 586–595 (2023). https://doi.org/10.1007/s12017-023-08761-2
Received:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1007/s12017-023-08761-2