PSEN1ΔE9, APPswe, and APOE4 Confer Disparate Phenotypes in Human iPSC-Derived Microglia
Henna Konttinen, Mauricio E Castro Cabral-da-Silva, Sohvi Ohtonen, Sara Wojciechowski, Anastasia Shakirzyanova, Simone Caligola, Rosalba Giugno, Yevheniia Ishchenko, Damián Hernández, Mohammad Feroze Fazaludeen, Shaila Eamen, Mireia Gómez Budia, Ilkka Fagerlund, Flavia Scoyni, Paula Korhonen, Nadine Huber, Annakaisa Haapasalo, Alex W Hewitt, James Vickers, Grady C Smith, Minna Oksanen, Caroline Graff, Katja M Kanninen, Sarka Lehtonen, Nicholas Propson, Michael P Schwartz, Alice Pébay, Jari Koistinaho, Lezanne Ooi, Tarja Malm, Henna Konttinen, Mauricio E Castro Cabral-da-Silva, Sohvi Ohtonen, Sara Wojciechowski, Anastasia Shakirzyanova, Simone Caligola, Rosalba Giugno, Yevheniia Ishchenko, Damián Hernández, Mohammad Feroze Fazaludeen, Shaila Eamen, Mireia Gómez Budia, Ilkka Fagerlund, Flavia Scoyni, Paula Korhonen, Nadine Huber, Annakaisa Haapasalo, Alex W Hewitt, James Vickers, Grady C Smith, Minna Oksanen, Caroline Graff, Katja M Kanninen, Sarka Lehtonen, Nicholas Propson, Michael P Schwartz, Alice Pébay, Jari Koistinaho, Lezanne Ooi, Tarja Malm
Abstract
Here we elucidate the effect of Alzheimer disease (AD)-predisposing genetic backgrounds, APOE4, PSEN1ΔE9, and APPswe, on functionality of human microglia-like cells (iMGLs). We present a physiologically relevant high-yield protocol for producing iMGLs from induced pluripotent stem cells. Differentiation is directed with small molecules through primitive erythromyeloid progenitors to re-create microglial ontogeny from yolk sac. The iMGLs express microglial signature genes and respond to ADP with intracellular Ca2+ release distinguishing them from macrophages. Using 16 iPSC lines from healthy donors, AD patients and isogenic controls, we reveal that the APOE4 genotype has a profound impact on several aspects of microglial functionality, whereas PSEN1ΔE9 and APPswe mutations trigger minor alterations. The APOE4 genotype impairs phagocytosis, migration, and metabolic activity of iMGLs but exacerbates their cytokine secretion. This indicates that APOE4 iMGLs are fundamentally unable to mount normal microglial functionality in AD.
Keywords: APOE; APPswe; Alzheimer disease; E9; PSEN1Δ; iPSC; metabolism; microglia; mitochondria; phagocytosis.
Copyright © 2019 The Author(s). Published by Elsevier Inc. All rights reserved.
Figures
References
- Abud E.M., Ramirez R.N., Martinez E.S., Healy L.M., Nguyen C.H.H., Newman S.A., Yeromin A.V., Scarfone V.M., Marsh S.E., Fimbres C. iPSC-derived human microglia-like cells to study neurological diseases. Neuron. 2017;94:278–293.e9.
- Bagyinszky E., Youn Y.C., An S.S., Kim S. The genetics of Alzheimer's disease. Clin. Interv. Aging. 2014;9:535–551.
- Balez R., Steiner N., Engel M., Munoz S.S., Lum J.S., Wu Y., Wang D., Vallotton P., Sachdev P., O'Connor M. Neuroprotective effects of apigenin against inflammation, neuronal excitability and apoptosis in an induced pluripotent stem cell model of Alzheimer's disease. Sci. Rep. 2016;6:31450.
- Banati R.B., Gehrmann J., Czech C., Monning U., Jones L.L., Konig G., Beyreuther K., Kreutzberg G.W. Early and rapid de novo synthesis of Alzheimer beta A4-amyloid precursor protein (APP) in activated microglia. Glia. 1993;9:199–210.
- Barroeta-Espar I., Weinstock L.D., Perez-Nievas B.G., Meltzer A.C., Siao Tick Chong M., Amaral A.C., Murray M.E., Moulder K.L., Morris J.C., Cairns N.J. Distinct cytokine profiles in human brains resilient to Alzheimer's pathology. Neurobiol. Dis. 2019;121:327–337.
- Bennett M.L., Bennett F.C., Liddelow S.A., Ajami B., Zamanian J.L., Fernhoff N.B., Mulinyawe S.B., Bohlen C.J., Adil A., Tucker A. New tools for studying microglia in the mouse and human CNS. Proc. Natl. Acad. Sci. U S A. 2016;113:E1738–E1746.
- Butovsky O., Jedrychowski M.P., Moore C.S., Cialic R., Lanser A.J., Gabriely G., Koeglsperger T., Dake B., Wu P.M., Doykan C.E. Identification of a unique TGF-beta-dependent molecular and functional signature in microglia. Nat. Neurosci. 2014;17:131–143.
- Caldeira C., Cunha C., Vaz A.R., Falcao A.S., Barateiro A., Seixas E., Fernandes A., Brites D. Key aging-associated alterations in primary microglia response to beta-amyloid stimulation. Front. Aging Neurosci. 2017;9:277.
- Colonna M., Butovsky O. Microglia function in the central nervous system during health and neurodegeneration. Annu. Rev. Immunol. 2017;35:441–468.
- Crombie D.E., Daniszewski M., Liang H.H., Kulkarni T., Li F., Lidgerwood G.E., Conquest A., Hernandez D., Hung S.S., Gill K.P. Development of a modular automated system for maintenance and differentiation of adherent human pluripotent stem cells. SLAS Discov. 2017;22:1016–1025.
- Crook R., Verkkoniemi A., Perez-Tur J., Mehta N., Baker M., Houlden H., Farrer M., Hutton M., Lincoln S., Hardy J. A variant of Alzheimer's disease with spastic paraparesis and unusual plaques due to deletion of exon 9 of presenilin 1. Nat. Med. 1998;4:452–455.
- De Simone R., Niturad C.E., De Nuccio C., Ajmone-Cat M.A., Visentin S., Minghetti L. TGF-beta and LPS modulate ADP-induced migration of microglial cells through P2Y1 and P2Y12 receptor expression. J. Neurochem. 2010;115:450–459.
- Douvaras P., Sun B., Wang M., Kruglikov I., Lallos G., Zimmer M., Terrenoire C., Zhang B., Gandy S., Schadt E. Directed differentiation of human pluripotent stem cells to Microglia. Stem Cell Reports. 2017;8:1516–1524.
- Engel M., Balez R., Munoz S.S., Cabral-da-Silva M.C., Stevens C.H., Bax M., Do-Ha D., Sidhu K., Sachdev P., Ooi L. Viral-free generation and characterization of a human induced pluripotent stem cell line from dermal fibroblasts. Stem Cell Res. 2018;32:135–138.
- Ghosh S., Castillo E., Frias E.S., Swanson R.A. Bioenergetic regulation of microglia. Glia. 2018;66:1200–1212.
- Ginhoux F., Greter M., Leboeuf M., Nandi S., See P., Gokhan S., Mehler M.F., Conway S.J., Ng L.G., Stanley E.R. Fate mapping analysis reveals that adult microglia derive from primitive macrophages. Science. 2010;330:841–845.
- Ginhoux F., Lim S., Hoeffel G., Low D., Huber T. Origin and differentiation of microglia. Front. Cell Neurosci. 2013;7:45.
- Haenseler W., Sansom S.N., Buchrieser J., Newey S.E., Moore C.S., Nicholls F.J., Chintawar S., Schnell C., Antel J.P., Allen N.D. A highly efficient human pluripotent stem cell Microglia model displays a neuronal-co-culture-specific expression profile and inflammatory response. Stem Cell Reports. 2017;8:1727–1742.
- Hoffmann A., Kann O., Ohlemeyer C., Hanisch U.K., Kettenmann H. Elevation of basal intracellular calcium as a central element in the activation of brain macrophages (microglia): suppression of receptor-evoked calcium signaling and control of release function. J. Neurosci. 2003;23:4410–4419.
- Holmqvist S., Lehtonen S., Chumarina M., Puttonen K.A., Azevedo C., Lebedeva O., Ruponen M., Oksanen M., Djelloul M., Collin A. Creation of a library of induced pluripotent stem cells from Parkinsonian patients. NPJ Parkinson's Dis. 2016;2:16009.
- Jayadev S., Case A., Alajajian B., Eastman A.J., Moller T., Garden G.A. Presenilin 2 influences miR146 level and activity in microglia. J. Neurochem. 2013;127:592–599.
- Kennedy M., D'Souza S.L., Lynch-Kattman M., Schwantz S., Keller G. Development of the hemangioblast defines the onset of hematopoiesis in human ES cell differentiation cultures. Blood. 2007;109:2679–2687.
- Kierdorf K., Erny D., Goldmann T., Sander V., Schulz C., Perdiguero E.G., Wieghofer P., Heinrich A., Riemke P., Holscher C. Microglia emerge from erythromyeloid precursors via Pu.1- and Irf8-dependent pathways. Nat. Neurosci. 2013;16:273–280.
- Koenigsknecht-Talboo J., Landreth G.E. Microglial phagocytosis induced by fibrillar beta-amyloid and IgGs are differentially regulated by proinflammatory cytokines. J. Neurosci. 2005;25:8240–8249.
- Krasemann S., Madore C., Cialic R., Baufeld C., Calcagno N., El Fatimy R., Beckers L., O'Loughlin E., Xu Y., Fanek Z. The TREM2-APOE pathway drives the transcriptional phenotype of dysfunctional microglia in neurodegenerative diseases. Immunity. 2017;47:566–581.e9.
- Lambert C., Ase A.R., Seguela P., Antel J.P. Distinct migratory and cytokine responses of human microglia and macrophages to ATP. Brain Behav. Immun. 2010;24:1241–1248.
- Lanzrein A.S., Johnston C.M., Perry V.H., Jobst K.A., King E.M., Smith A.D. Longitudinal study of inflammatory factors in serum, cerebrospinal fluid, and brain tissue in Alzheimer disease: interleukin-1beta, interleukin-6, interleukin-1 receptor antagonist, tumor necrosis factor-alpha, the soluble tumor necrosis factor receptors I and II, and alpha1-antichymotrypsin. Alzheimer Dis. Assoc. Disord. 1998;12:215–227.
- Lavin Y., Winter D., Blecher-Gonen R., David E., Keren-Shaul H., Merad M., Jung S., Amit I. Tissue-resident macrophage enhancer landscapes are shaped by the local microenvironment. Cell. 2014;159:1312–1326.
- Lee C.Y., Landreth G.E. The role of microglia in amyloid clearance from the AD brain. J. Neural Transm. (Vienna) 2010;117:949–960.
- Lin Y.T., Seo J., Gao F., Feldman H.M., Wen H.L., Penney J., Cam H.P., Gjoneska E., Raja W.K., Cheng J. APOE4 causes widespread molecular and cellular alterations associated with Alzheimer's disease phenotypes in human iPSC-derived brain cell types. Neuron. 2018;98:1141–1154.e7.
- Liu C.C., Liu C.C., Kanekiyo T., Xu H., Bu G. Apolipoprotein E and Alzheimer disease: risk, mechanisms and therapy. Nat. Rev. Neurol. 2013;9:106–118.
- Manocha G.D., Floden A.M., Rausch K., Kulas J.A., McGregor B.A., Rojanathammanee L., Puig K.R., Puig K.L., Karki S., Nichols M.R. APP regulates microglial phenotype in a mouse model of Alzheimer's disease. J. Neurosci. 2016;36:8471–8486.
- McCaughey T., Liang H.H., Chen C., Fenwick E., Rees G., Wong R.C., Vickers J.C., Summers M.J., MacGregor C., Craig J.E. An interactive multimedia approach to improving informed consent for induced pluripotent stem cell research. Cell Stem Cell. 2016;18:307–308.
- McQuade A., Coburn M., Tu C.H., Hasselmann J., Davtyan H., Blurton-Jones M. Development and validation of a simplified method to generate human microglia from pluripotent stem cells. Mol. Neurodegener. 2018;13:67.
- Muffat J., Li Y., Yuan B., Mitalipova M., Omer A., Corcoran S., Bakiasi G., Tsai L.H., Aubourg P., Ransohoff R.M. Efficient derivation of microglia-like cells from human pluripotent stem cells. Nat. Med. 2016;22:1358–1367.
- Mullan M., Crawford F., Axelman K., Houlden H., Lilius L., Winblad B., Lannfelt L. A pathogenic mutation for probable Alzheimer's disease in the APP gene at the N-terminus of beta-amyloid. Nat. Genet. 1992;1:345–347.
- Munoz S.S., Balez R., Castro Cabral-da-Silva M.E., Berg T., Engel M., Bax M., Do-Ha D., Stevens C.H., Greenough M., Bush A. Generation and characterization of human induced pluripotent stem cell lines from a familial Alzheimer's disease PSEN1 A246E patient and a non-demented family member bearing wild-type PSEN1. Stem Cell Res. 2018;31:227–230.
- Nadler Y., Alexandrovich A., Grigoriadis N., Hartmann T., Rao K.S., Shohami E., Stein R. Increased expression of the gamma-secretase components presenilin-1 and nicastrin in activated astrocytes and microglia following traumatic brain injury. Glia. 2008;56:552–567.
- Oberstein T.J., Spitzer P., Klafki H.W., Linning P., Neff F., Knolker H.J., Lewczuk P., Wiltfang J., Kornhuber J., Maler J.M. Astrocytes and microglia but not neurons preferentially generate N-terminally truncated Abeta peptides. Neurobiol. Dis. 2015;73:24–35.
- Okita K., Matsumura Y., Sato Y., Okada A., Morizane A., Okamoto S., Hong H., Nakagawa M., Tanabe K., Tezuka K. A more efficient method to generate integration-free human iPS cells. Nat. Methods. 2011;8:409–412.
- Oksanen M., Hyotylainen I., Voutilainen J., Puttonen K.A., Hamalainen R.H., Graff C., Lehtonen S., Koistinaho J. Generation of a human induced pluripotent stem cell line (LL008 1.4) from a familial Alzheimer's disease patient carrying a double KM670/671NL (Swedish) mutation in APP gene. Stem Cell Res. 2018;31:181–185.
- Oksanen M., Petersen A.J., Naumenko N., Puttonen K., Lehtonen S., Gubert Olive M., Shakirzyanova A., Leskela S., Sarajarvi T., Viitanen M. PSEN1 mutant iPSC-derived model reveals severe astrocyte pathology in Alzheimer's disease. Stem Cell Reports. 2017;9:1885–1897.
- Olah M., Patrick E., Villani A.C., Xu J., White C.C., Ryan K.J., Piehowski P., Kapasi A., Nejad P., Cimpean M. A transcriptomic atlas of aged human microglia. Nat. Commun. 2018;9:539.
- Ooi L., Sidhu K., Poljak A., Sutherland G., O'Connor M.D., Sachdev P., Munch G. Induced pluripotent stem cells as tools for disease modelling and drug discovery in Alzheimer's disease. J. Neural Transm. (Vienna) 2013;120:103–111.
- Orihuela R., McPherson C.A., Harry G.J. Microglial M1/M2 polarization and metabolic states. Br. J. Pharmacol. 2016;173:649–665.
- Pandya H., Shen M.J., Ichikawa D.M., Sedlock A.B., Choi Y., Johnson K.R., Kim G., Brown M.A., Elkahloun A.G., Maric D. Differentiation of human and murine induced pluripotent stem cells to microglia-like cells. Nat. Neurosci. 2017;20:753–759.
- Pulido-Salgado M., Vidal-Taboada J.M., Barriga G.G., Sola C., Saura J. RNA-Seq transcriptomic profiling of primary murine microglia treated with LPS or LPS + IFNgamma. Sci. Rep. 2018;8:16096.
- Qu C., Puttonen K.A., Lindeberg H., Ruponen M., Hovatta O., Koistinaho J., Lammi M.J. Chondrogenic differentiation of human pluripotent stem cells in chondrocyte co-culture. Int. J. Biochem. Cell Biol. 2013;45:1802–1812.
- Rustenhoven J., Park T.I., Schweder P., Scotter J., Correia J., Smith A.M., Gibbons H.M., Oldfield R.L., Bergin P.S., Mee E.W. Isolation of highly enriched primary human microglia for functional studies. Sci. Rep. 2016;6:19371.
- Saijo K., Glass C.K. Microglial cell origin and phenotypes in health and disease. Nat. Rev. Immunol. 2011;11:775–787.
- Scheuner D., Eckman C., Jensen M., Song X., Citron M., Suzuki N., Bird T.D., Hardy J., Hutton M., Kukull W. Secreted amyloid beta-protein similar to that in the senile plaques of Alzheimer's disease is increased in vivo by the presenilin 1 and 2 and APP mutations linked to familial Alzheimer's disease. Nat. Med. 1996;2:864–870.
- Schwartz M.P., Hou Z., Propson N.E., Zhang J., Engstrom C.J., Santos Costa V., Jiang P., Nguyen B.K., Bolin J.M., Daly W. Human pluripotent stem cell-derived neural constructs for predicting neural toxicity. Proc. Natl. Acad. Sci. U S A. 2015;112:12516–12521.
- Selkoe D.J. The cell biology of beta-amyloid precursor protein and presenilin in Alzheimer's disease. Trends Cell Biol. 1998;8:447–453.
- Shi Y., Holtzman D.M. Interplay between innate immunity and Alzheimer disease: APOE and TREM2 in the spotlight. Nat. Rev. Immunol. 2018;18:759–772.
- Smith J.A., Das A., Ray S.K., Banik N.L. Role of pro-inflammatory cytokines released from microglia in neurodegenerative diseases. Brain Res. Bull. 2012;87:10–20.
- Sturgeon C.M., Ditadi A., Awong G., Kennedy M., Keller G. Wnt signaling controls the specification of definitive and primitive hematopoiesis from human pluripotent stem cells. Nat. Biotechnol. 2014;32:554–561.
- Ta T.T., Dikmen H.O., Schilling S., Chausse B., Lewen A., Hollnagel J.O., Kann O. Priming of microglia with IFN-gamma slows neuronal gamma oscillations in situ. Proc. Natl. Acad. Sci. U S A. 2019
- Townsend K.P., Town T., Mori T., Lue L.F., Shytle D., Sanberg P.R., Morgan D., Fernandez F., Flavell R.A., Tan J. CD40 signaling regulates innate and adaptive activation of microglia in response to amyloid beta-peptide. Eur. J. Immunol. 2005;35:901–910.
- Uenishi G., Theisen D., Lee J.H., Kumar A., Raymond M., Vodyanik M., Swanson S., Stewart R., Thomson J., Slukvin I. Tenascin C promotes hematoendothelial development and T lymphoid commitment from human pluripotent stem cells in chemically defined conditions. Stem Cell Reports. 2014;3:1073–1084.
- Ulland T.K., Song W.M., Huang S.C., Ulrich J.D., Sergushichev A., Beatty W.L., Loboda A.A., Zhou Y., Cairns N.J., Kambal A. TREM2 maintains microglial metabolic fitness in Alzheimer's disease. Cell. 2017;170:649–663.e13.
- Wang W.Y., Tan M.S., Yu J.T., Tan L. Role of pro-inflammatory cytokines released from microglia in Alzheimer's disease. Ann. Transl. Med. 2015;3:136.
- Weuve J., Hebert L.E., Scherr P.A., Evans D.A. Deaths in the United States among persons with Alzheimer's disease (2010-2050) Alzheimers Dement. 2014;10:e40–e46.
- Xu M., Zhang L., Liu G., Jiang N., Zhou W., Zhang Y. Pathological changes in alzheimer's disease analyzed using induced pluripotent stem cell-derived human Microglia-like cells. J. Alzheimers Dis. 2019;67:357–368.
- Zhang Y., Chen K., Sloan S.A., Bennett M.L., Scholze A.R., O'Keeffe S., Phatnani H.P., Guarnieri P., Caneda C., Ruderisch N. An RNA-sequencing transcriptome and splicing database of glia, neurons, and vascular cells of the cerebral cortex. J. Neurosci. 2014;34:11929–11947.
- Zhao Y., Li X., Huang T., Jiang L.L., Tan Z., Zhang M., Cheng I.H., Wang X., Bu G., Zhang Y.W. Intracellular trafficking of TREM2 is regulated by presenilin 1. Exp. Mol. Med. 2017;49:e405.
Source: PubMed