臺灣博碩士論文加值系統

阿茲海默氏症是最常見的失智症形式，其特徵是記憶和認知功能的進行性退化，影響70％的失智症患者。流行病學和臨床數據顯示，血清中高濃度的維生素D與更好的認知能力和神經保護功能有所關聯，而維生素D缺乏是一個相當嚴重的結果，並且可以在許多疾病中觀察到，尤其是阿茲海默氏症。因此，我們假設補充維生素D可能具有延緩阿茲海默氏症病程發展的治療效果。在這項研究中，我們在3xTg-AD小鼠飼料中加入維生素D補充劑，共餵食4個月的時間，發現維生素D補充劑顯著地增加維生素D濃度並改善3xTg-AD小鼠的認知缺陷。先前的研究表明CRMP2磷酸化的上升是阿茲海默氏症的特徵。我們發現補充維生素D可以減少3xTg-AD小鼠海馬迴中CRMP2的磷酸化表達，而使用蛋白磷酸酶1和2A的抑製劑岡田酸（OA）微量注射到3xTg-AD的海馬迴中可以重新活化CRMP2磷酸化並且使餵食維生素D的3xTg-AD小鼠組別再次出現記憶障礙。此外，最近的研究表示，GSK-3β抑製劑SB216763和SB415286可以調節CRMP2磷酸化。注射SB216763或SB415286後，我們發現與對照組相比，GSK-3β抑製劑可降低3xTg-AD小鼠的CRMP2磷酸化並改善空間記憶，進一步證實CRMP2磷酸化能誘導阿茲海默氏症的記憶障礙。此外，許多證據表明CRMP2可能與NMDAR的NR2B亞基相互作用。免疫沉澱法顯示在與對照治療組相比下，維生素D補充劑可以逆轉CRMP2-NR2B相互作用。TAT-CBD3破壞CRMP2-NR2B相互作用進而抑制由維生素D誘導的記憶改善。綜合以上研究結果，本研究希望對阿茲海默氏症治療，預防和治療策略上做出有價值的貢獻。

Alzheimer's disease (AD) is the most common form of dementia, affecting up to 70% of all people with dementia and characterized by progressive exacerbation of memory and cognitive function. Epidemiological and clinical data show that high serum levels of vitamin D links with better cognitive performance and neuroprotective functions, whereas the consequences of vitamin D deficiency are severe and can be observed in many diseases progression, especially AD. Thus, we hypothesized that vitamin D supplementation may have therapeutic efficacy to slow down the AD progression. In this study, we fed 3xTg-AD mice with vitamin D-supplemented chow for four months and found that vitamin D supplementation significantly increased vitamin D concentrations and improved cognitive deficits in 3xTg-AD mice. Previous studies indicated that increased collapsin response mediator protein 2 (CRMP2) phosphorylation is characteristic of AD. We also found that vitamin D supplementation could decrease phosphorylated CRMP2 expression in the hippocampus of 3xTg-AD mice, whereas reactivation of CRMP2 phosphorylation by using microinjection of okadaic acid (OA), an inhibitor of protein phosphatases 1 and 2A, into the hippocampus of 3xTg-AD mice with vitamin D diet exhibited memory impairment. In addition, recent studies demonstrated that glycogen synthase kinase 3β (GSK-3β) inhibitors SB216763 and SB415286 could regulate CRMP2 phosphorylation. After injection of SB216763 or SB415286, we found that GSK-3β inhibitors could decrease pCRMP2 expressions and improved spatial memory in 3xTg-AD mice compared to that of vehicle control, confirming that increased pCRMP2 triggered memory impairment. Moreover, evidence suggested that CRMP2 could interact with the NR2B subunit of NMDAR. Immunoprecipitation analysis showed that vitamin D reversed CRMP2-NR2B interaction compare to that of control diet-treated groups, whereas disruption of the CRMP2-NR2B interaction using TAT-CBD3 peptide abolished vitamin D-mediated memory improvement. Taken together, this study should make a valuable contribution to the development of promising strategies for management, prevention, and treatment of AD.

Abstract in Chinese I
Abstract in English III
Content VII
List of Figures IX
Abbreviations XI
Chapter 1 Introduction 1
Chapter 2 Specific Aims 8
Chapter 3 Materials and methods 10
Chapter 4 Results 18
Chapter 5 Discussion 44
References 48

Afzal, S., S. E. Bojesen, and B. G. Nordestgaard. 2014. 'Reduced 25-hydroxyvitamin D and risk of Alzheimer's disease and vascular dementia', Alzheimers Dement, 10: 296-302.
Al-Hallaq, R. A., T. P. Conrads, T. D. Veenstra, and R. J. Wenthold. 2007. 'NMDA di-heteromeric receptor populations and associated proteins in rat hippocampus', J Neurosci, 27: 8334-43.
Alzheimer's, Association. 2016. '2016 Alzheimer's disease facts and figures', Alzheimers Dement, 12: 459-509.
Anastasiou, C. A., M. Yannakoulia, and N. Scarmeas. 2014. 'Vitamin D and cognition: an update of the current evidence', J Alzheimers Dis, 42 Suppl 3: S71-80.
Andel, R., K. Hyer, and A. Slack. 2007. 'Risk factors for nursing home placement in older adults with and without dementia', J Aging Health, 19: 213-28.
Annweiler, C., D. J. Llewellyn, and O. Beauchet. 2013. 'Low serum vitamin D concentrations in Alzheimer's disease: a systematic review and meta-analysis', J Alzheimers Dis, 33: 659-74.
Baker, S., and J. Gotz. 2016. 'A local insult of okadaic acid in wild-type mice induces tau phosphorylation and protein aggregation in anatomically distinct brain regions', Acta Neuropathol Commun, 4: 32.
Birks, J. 2006. 'Cholinesterase inhibitors for Alzheimer's disease', Cochrane Database Syst Rev: CD005593.
Bloom, G. S. 2014. 'Amyloid-beta and tau: the trigger and bullet in Alzheimer disease pathogenesis', JAMA Neurol, 71: 505-8.
Brittain, J. M., D. B. Duarte, S. M. Wilson, W. Zhu, C. Ballard, P. L. Johnson, N. Liu, W. Xiong, M. S. Ripsch, Y. Wang, J. C. Fehrenbacher, S. D. Fitz, M. Khanna, C. K. Park, B. S. Schmutzler, B. M. Cheon, M. R. Due, T. Brustovetsky, N. M. Ashpole, A. Hudmon, S. O. Meroueh, C. M. Hingtgen, N. Brustovetsky, R. R. Ji, J. H. Hurley, X. Jin, A. Shekhar, X. M. Xu, G. S. Oxford, M. R. Vasko, F. A. White, and R. Khanna. 2011. 'Suppression of inflammatory and neuropathic pain by uncoupling CRMP-2 from the presynaptic Ca(2)(+) channel complex', Nat Med, 17: 822-9.
Brittain, J. M., R. Pan, H. You, T. Brustovetsky, N. Brustovetsky, G. W. Zamponi, W. H. Lee, and R. Khanna. 2012. 'Disruption of NMDAR-CRMP-2 signaling protects against focal cerebral ischemic damage in the rat middle cerebral artery occlusion model', Channels (Austin), 6: 52-9.
Brittain, J. M., A. D. Piekarz, Y. Wang, T. Kondo, T. R. Cummins, and R. Khanna. 2009. 'An atypical role for collapsin response mediator protein 2 (CRMP-2) in neurotransmitter release via interaction with presynaptic voltage-gated calcium channels', J Biol Chem, 284: 31375-90.
Brustovetsky, T., J. J. Pellman, X. F. Yang, R. Khanna, and N. Brustovetsky. 2014. 'Collapsin response mediator protein 2 (CRMP2) interacts with N-methyl-D-aspartate (NMDA) receptor and Na+/Ca2+ exchanger and regulates their functional activity', J Biol Chem, 289: 7470-82.
Burns, A., and S. Iliffe. 2009. 'Alzheimer's disease', BMJ, 338: b158.
Cao, X., Z. Cui, R. Feng, Y. P. Tang, Z. Qin, B. Mei, and J. Z. Tsien. 2007. 'Maintenance of superior learning and memory function in NR2B transgenic mice during ageing', Eur J Neurosci, 25: 1815-22.
Charrier, E., S. Reibel, V. Rogemond, M. Aguera, N. Thomasset, and J. Honnorat. 2003. 'Collapsin response mediator proteins (CRMPs): involvement in nervous system development and adult neurodegenerative disorders', Mol Neurobiol, 28: 51-64.
Chen, H., W. Xue, J. Li, K. Fu, H. Shi, B. Zhang, W. Teng, and L. Tian. 2018. '25-Hydroxyvitamin D Levels and the Risk of Dementia and Alzheimer's Disease: A Dose-Response Meta-Analysis', Front Aging Neurosci, 10: 368.
Clare, L., R. T. Woods, E. D. Moniz Cook, M. Orrell, and A. Spector. 2003. 'Cognitive rehabilitation and cognitive training for early-stage Alzheimer's disease and vascular dementia', Cochrane Database Syst Rev: CD003260.
Cole, A. R., F. Causeret, G. Yadirgi, C. J. Hastie, H. McLauchlan, E. J. McManus, F. Hernandez, B. J. Eickholt, M. Nikolic, and C. Sutherland. 2006. 'Distinct priming kinases contribute to differential regulation of collapsin response mediator proteins by glycogen synthase kinase-3 in vivo', J Biol Chem, 281: 16591-8.
Cole, A. R., W. Noble, L. van Aalten, F. Plattner, R. Meimaridou, D. Hogan, M. Taylor, J. LaFrancois, F. Gunn-Moore, A. Verkhratsky, S. Oddo, F. LaFerla, K. P. Giese, K. T. Dineley, K. Duff, J. C. Richardson, S. D. Yan, D. P. Hanger, S. M. Allan, and C. Sutherland. 2007. 'Collapsin response mediator protein-2 hyperphosphorylation is an early event in Alzheimer's disease progression', J Neurochem, 103: 1132-44.
Corcoran, K. A., K. Leaderbrand, V. Jovasevic, A. L. Guedea, F. Kassam, and J. Radulovic. 2015. 'Regulation of fear extinction versus other affective behaviors by discrete cortical scaffolding complexes associated with NR2B and PKA signaling', Transl Psychiatry, 5: e657.
DeLuca, H. F. 2004. 'Overview of general physiologic features and functions of vitamin D', Am J Clin Nutr, 80: 1689S-96S.
Demay, M. B. 2018. 'The good and the bad of vitamin D inactivation', J Clin Invest, 128: 3736-38.
Devanand, D. P., D. M. Jacobs, M. X. Tang, C. Del Castillo-Castaneda, M. Sano, K. Marder, K. Bell, F. W. Bylsma, J. Brandt, M. Albert, and Y. Stern. 1997. 'The course of psychopathologic features in mild to moderate Alzheimer disease', Arch Gen Psychiatry, 54: 257-63.
Disease, G. B. D., Incidence Injury, and Collaborators Prevalence. 2016. 'Global, regional, and national incidence, prevalence, and years lived with disability for 310 diseases and injuries, 1990-2015: a systematic analysis for the Global Burden of Disease Study 2015', Lancet, 388: 1545-602.
Dubois, B., H. Hampel, H. H. Feldman, P. Scheltens, P. Aisen, S. Andrieu, H. Bakardjian, H. Benali, L. Bertram, K. Blennow, K. Broich, E. Cavedo, S. Crutch, J. F. Dartigues, C. Duyckaerts, S. Epelbaum, G. B. Frisoni, S. Gauthier, R. Genthon, A. A. Gouw, M. O. Habert, D. M. Holtzman, M. Kivipelto, S. Lista, J. L. Molinuevo, S. E. O'Bryant, G. D. Rabinovici, C. Rowe, S. Salloway, L. S. Schneider, R. Sperling, M. Teichmann, M. C. Carrillo, J. Cummings, C. R. Jack, Jr., Group Proceedings of the Meeting of the International Working, A. D. the American Alzheimer's Association on The Preclinical State of, July, and U. S. A. Washington Dc. 2016. 'Preclinical Alzheimer's disease: Definition, natural history, and diagnostic criteria', Alzheimers Dement, 12: 292-323.
Dubois, B., A. Padovani, P. Scheltens, A. Rossi, and G. Dell'Agnello. 2016. 'Timely Diagnosis for Alzheimer's Disease: A Literature Review on Benefits and Challenges', J Alzheimers Dis, 49: 617-31.
Eldar-Finkelman, H., and A. Martinez. 2011. 'GSK-3 Inhibitors: Preclinical and Clinical Focus on CNS', Front Mol Neurosci, 4: 32.
Evatt, M. L., M. R. Delong, N. Khazai, A. Rosen, S. Triche, and V. Tangpricha. 2008. 'Prevalence of vitamin d insufficiency in patients with Parkinson disease and Alzheimer disease', Arch Neurol, 65: 1348-52.
Fischer, G., B. Pan, D. Vilceanu, Q. H. Hogan, and H. Yu. 2014. 'Sustained relief of neuropathic pain by AAV-targeted expression of CBD3 peptide in rat dorsal root ganglion', Gene Ther, 21: 44-51.
Fukata, Y., T. J. Itoh, T. Kimura, C. Menager, T. Nishimura, T. Shiromizu, H. Watanabe, N. Inagaki, A. Iwamatsu, H. Hotani, and K. Kaibuchi. 2002. 'CRMP-2 binds to tubulin heterodimers to promote microtubule assembly', Nat Cell Biol, 4: 583-91.
Garcion, E., N. Wion-Barbot, C. N. Montero-Menei, F. Berger, and D. Wion. 2002. 'New clues about vitamin D functions in the nervous system', Trends Endocrinol Metab, 13: 100-5.
Gold, C. A., and A. E. Budson. 2008. 'Memory loss in Alzheimer's disease: implications for development of therapeutics', Expert Rev Neurother, 8: 1879-91.
Goshima, Y., F. Nakamura, P. Strittmatter, and S. M. Strittmatter. 1995. 'Collapsin-induced growth cone collapse mediated by an intracellular protein related to UNC-33', Nature, 376: 509-14.
Hewison, M. 2011. 'Vitamin D and innate and adaptive immunity', Vitam Horm, 86: 23-62.
Hsu, D., and G. A. Marshall. 2017. 'Primary and Secondary Prevention Trials in Alzheimer Disease: Looking Back, Moving Forward', Curr Alzheimer Res, 14: 426-40.
Hu, S., A. N. Begum, M. R. Jones, M. S. Oh, W. K. Beech, B. H. Beech, F. Yang, P. Chen, O. J. Ubeda, P. C. Kim, P. Davies, Q. Ma, G. M. Cole, and S. A. Frautschy. 2009. 'GSK3 inhibitors show benefits in an Alzheimer's disease (AD) model of neurodegeneration but adverse effects in control animals', Neurobiol Dis, 33: 193-206.
Iacopetta, K., L. E. Collins-Praino, F. T. A. Buisman-Pijlman, J. Liu, A. D. Hutchinson, and M. R. Hutchinson. 2018. 'Are the protective benefits of vitamin D in neurodegenerative disease dependent on route of administration? A systematic review', Nutr Neurosci: 1-30.
Inagaki, N., K. Chihara, N. Arimura, C. Menager, Y. Kawano, N. Matsuo, T. Nishimura, M. Amano, and K. Kaibuchi. 2001. 'CRMP-2 induces axons in cultured hippocampal neurons', Nat Neurosci, 4: 781-2.
Ittner, L. M., and J. Gotz. 2011. 'Amyloid-beta and tau--a toxic pas de deux in Alzheimer's disease', Nat Rev Neurosci, 12: 65-72.
Jahn, H. 2013. 'Memory loss in Alzheimer's disease', Dialogues Clin Neurosci, 15: 445-54.
Kamat, P. K., S. Rai, and C. Nath. 2013. 'Okadaic acid induced neurotoxicity: an emerging tool to study Alzheimer's disease pathology', Neurotoxicology, 37: 163-72.
Kaplan, I. M., J. S. Wadia, and S. F. Dowdy. 2005. 'Cationic TAT peptide transduction domain enters cells by macropinocytosis', J Control Release, 102: 247-53.
Keeney, J. T., and D. A. Butterfield. 2015. 'Vitamin D deficiency and Alzheimer disease: Common links', Neurobiol Dis, 84: 84-98.
Kennel, K. A., M. T. Drake, and D. L. Hurley. 2010. 'Vitamin D deficiency in adults: when to test and how to treat', Mayo Clin Proc, 85: 752-7; quiz 57-8.
Kimura, T., H. Watanabe, A. Iwamatsu, and K. Kaibuchi. 2005. 'Tubulin and CRMP-2 complex is transported via Kinesin-1', J Neurochem, 93: 1371-82.
King, M. K., M. Pardo, Y. Cheng, K. Downey, R. S. Jope, and E. Beurel. 2014. 'Glycogen synthase kinase-3 inhibitors: Rescuers of cognitive impairments', Pharmacol Ther, 141: 1-12.
Landel, V., C. Annweiler, P. Millet, M. Morello, and F. Feron. 2016. 'Vitamin D, Cognition and Alzheimer's Disease: The Therapeutic Benefit is in the D-Tails', J Alzheimers Dis, 53: 419-44.
Landel, V., P. Millet, K. Baranger, B. Loriod, and F. Feron. 2016. 'Vitamin D interacts with Esr1 and Igf1 to regulate molecular pathways relevant to Alzheimer's disease', Mol Neurodegener, 11: 22.
Littlejohns, T. J., W. E. Henley, I. A. Lang, C. Annweiler, O. Beauchet, P. H. Chaves, L. Fried, B. R. Kestenbaum, L. H. Kuller, K. M. Langa, O. L. Lopez, K. Kos, M. Soni, and D. J. Llewellyn. 2014. 'Vitamin D and the risk of dementia and Alzheimer disease', Neurology, 83: 920-8.
Lukaszyk, E., K. Bien-Barkowska, and B. Bien. 2018. 'Cognitive Functioning of Geriatric Patients: Is Hypovitaminosis D the Next Marker of Cognitive Dysfunction and Dementia?', Nutrients, 10.
Mazahery, H., and P. R. von Hurst. 2015. 'Factors Affecting 25-Hydroxyvitamin D Concentration in Response to Vitamin D Supplementation', Nutrients, 7: 5111-42.
Munday, R. 2013. 'Is protein phosphatase inhibition responsible for the toxic effects of okadaic Acid in animals?', Toxins (Basel), 5: 267-85.
Nagai, J., K. Owada, Y. Kitamura, Y. Goshima, and T. Ohshima. 2016. 'Inhibition of CRMP2 phosphorylation repairs CNS by regulating neurotrophic and inhibitory responses', Exp Neurol, 277: 283-95.
Nishimura, T., Y. Fukata, K. Kato, T. Yamaguchi, Y. Matsuura, H. Kamiguchi, and K. Kaibuchi. 2003. 'CRMP-2 regulates polarized Numb-mediated endocytosis for axon growth', Nat Cell Biol, 5: 819-26.
Olsson, E., L. Byberg, B. Karlstrom, T. Cederholm, H. Melhus, P. Sjogren, and L. Kilander. 2017. 'Vitamin D is not associated with incident dementia or cognitive impairment: an 18-y follow-up study in community-living old men', Am J Clin Nutr, 105: 936-43.
Ouma, S., M. Suenaga, F. F. Bolukbasi Hatip, I. Hatip-Al-Khatib, Y. Tsuboi, and Y. Matsunaga. 2018. 'Serum vitamin D in patients with mild cognitive impairment and Alzheimer's disease', Brain Behav, 8: e00936.
Perry, R. J., and J. R. Hodges. 1999. 'Attention and executive deficits in Alzheimer's disease. A critical review', Brain, 122 ( Pt 3): 383-404.
Reichrath, J., and B. Nurnberg. 2009. 'Cutaneous vitamin D synthesis versus skin cancer development: The Janus faces of solar UV-radiation', Dermatoendocrinol, 1: 253-61.
Reisberg, B., R. Doody, A. Stoffler, F. Schmitt, S. Ferris, H. J. Mobius, and Group Memantine Study. 2003. 'Memantine in moderate-to-severe Alzheimer's disease', N Engl J Med, 348: 1333-41.
Soutar, M. P., P. Thornhill, A. R. Cole, and C. Sutherland. 2009. 'Increased CRMP2 phosphorylation is observed in Alzheimer's disease; does this tell us anything about disease development?', Curr Alzheimer Res, 6: 269-78.
Spires-Jones, T. L., and B. T. Hyman. 2014. 'The intersection of amyloid beta and tau at synapses in Alzheimer's disease', Neuron, 82: 756-71.
Stenmark, P., D. Ogg, S. Flodin, A. Flores, T. Kotenyova, T. Nyman, P. Nordlund, and P. Kursula. 2007. 'The structure of human collapsin response mediator protein 2, a regulator of axonal growth', J Neurochem, 101: 906-17.
Tarawneh, R., and D. M. Holtzman. 2012. 'The clinical problem of symptomatic Alzheimer disease and mild cognitive impairment', Cold Spring Harb Perspect Med, 2: a006148.
Uchida, Y., T. Ohshima, Y. Sasaki, H. Suzuki, S. Yanai, N. Yamashita, F. Nakamura, K. Takei, Y. Ihara, K. Mikoshiba, P. Kolattukudy, J. Honnorat, and Y. Goshima. 2005. 'Semaphorin3A signalling is mediated via sequential Cdk5 and GSK3beta phosphorylation of CRMP2: implication of common phosphorylating mechanism underlying axon guidance and Alzheimer's disease', Genes Cells, 10: 165-79.
Wang, D., S. A. Jacobs, and J. Z. Tsien. 2014. 'Targeting the NMDA receptor subunit NR2B for treating or preventing age-related memory decline', Expert Opin Ther Targets, 18: 1121-30.
Weuve, J., L. E. Hebert, P. A. Scherr, and D. A. Evans. 2014. 'Deaths in the United States among persons with Alzheimer's disease (2010-2050)', Alzheimers Dement, 10: e40-6.
Williamson, R., L. van Aalten, D. M. Mann, B. Platt, F. Plattner, L. Bedford, J. Mayer, D. Howlett, A. Usardi, C. Sutherland, and A. R. Cole. 2011. 'CRMP2 hyperphosphorylation is characteristic of Alzheimer's disease and not a feature common to other neurodegenerative diseases', J Alzheimers Dis, 27: 615-25.
Wrzosek, M., J. Lukaszkiewicz, M. Wrzosek, A. Jakubczyk, H. Matsumoto, P. Piatkiewicz, M. Radziwon-Zaleska, M. Wojnar, and G. Nowicka. 2013. 'Vitamin D and the central nervous system', Pharmacol Rep, 65: 271-8.
Yamashita, N., and Y. Goshima. 2012. 'Collapsin response mediator proteins regulate neuronal development and plasticity by switching their phosphorylation status', Mol Neurobiol, 45: 234-46.
Yoshida, H., A. Watanabe, and Y. Ihara. 1998. 'Collapsin response mediator protein-2 is associated with neurofibrillary tangles in Alzheimer's disease', J Biol Chem, 273: 9761-8.
Yoshimura, T., Y. Kawano, N. Arimura, S. Kawabata, A. Kikuchi, and K. Kaibuchi. 2005. 'GSK-3beta regulates phosphorylation of CRMP-2 and neuronal polarity', Cell, 120: 137-49.
Yu, J., M. Gattoni-Celli, H. Zhu, N. R. Bhat, K. Sambamurti, S. Gattoni-Celli, and M. S. Kindy. 2011. 'Vitamin D3-enriched diet correlates with a decrease of amyloid plaques in the brain of AbetaPP transgenic mice', J Alzheimers Dis, 25: 295-307.