iPSC-Derived PSEN2 (N141I) Astrocytes and Microglia Exhibit a Primed Inflammatory Phenotype

Background Widescale evidence points to the involvement of glia and immune pathways in the progression of Alzheimer’s disease (AD). AD-associated iPSC-derived glial cells show a diverse range of AD-related phenotypic states encompassing cytokine/chemokine release, phagocytosis and morphological profiles, but to date studies are limited to cells derived from PSEN1, APOE and APP mutations or sporadic patients. The aim of the current study was to successfully differentiate iPSC-derived microglia and astrocytes from patients harbouring an AD-causative PSEN2 (N141I) mutation and characterise the inflammatory and morphological profile of these cells. Methods iPSCs from three healthy control individuals and three familial AD patients harbouring a heterozygous PSEN2 (N141I) mutation were used to derive astrocytes and microglia-like cells and cell identity and morphology were characterised through immunofluorescent microscopy. Cellular characterisation involved the stimulation of these cells by LPS and Aβ42 and analysis of cytokine/chemokine release was conducted through ELISAs and multi-cytokine arrays. The phagocytic capacity of these cells was then indexed by the uptake of fluorescently labelled fibrillar Aβ42. Results AD-derived astrocytes and microglia-like cells exhibited an atrophied and less complex morphological appearance than healthy controls. AD-derived astrocytes showed increased basal expression of GFAP, S100β and increased secretion and phagocytosis of Aβ42 while AD-derived microglia-like cells showed decreased IL-8 secretion compared to healthy controls. Upon immunological challenge AD-derived astrocytes and microglia-like cells show exaggerated secretion of the pro-inflammatory IL-6, CXCL1, ICAM-1 and IL-8 from astrocytes and IL-18 and MIF from microglia. Conclusion Our study showed, for the first time, the differentiation and characterisation of iPSC-derived astrocytes and microglia-like cells harbouring a PSEN2 (N141I) mutation. PSEN2 (N141I)-mutant astrocytes and microglia-like cells presented with a ‘primed’ phenotype characterised by reduced morphological complexity, exaggerated pro-inflammatory cytokine secretion and altered Aβ42 production and phagocytosis.

[1]  D. Boche,et al.  Microglial morphology in Alzheimer’s disease and after Aβ immunotherapy , 2021, Scientific Reports.

[2]  K. Mackie,et al.  Tubular human brain organoids to model microglia-mediated neuroinflammation. , 2021, Lab on a chip.

[3]  C. Webber,et al.  TREM2 Alzheimer’s variant R47H causes similar transcriptional dysregulation to knockout, yet only subtle functional phenotypes in human iPSC-derived macrophages , 2020, Alzheimer's research & therapy.

[4]  Katherine E. Prater,et al.  Early-Onset Familial Alzheimer Disease Variant PSEN2 N141I Heterozygosity is Associated with Altered Microglia Phenotype , 2020, Journal of Alzheimer's disease : JAD.

[5]  R. Lanzenberger,et al.  Dysfunction of the Blood-Brain Barrier—A Key Step in Neurodegeneration and Dementia , 2020, Frontiers in Aging Neuroscience.

[6]  Melanie A. Huntley,et al.  Alzheimer’s Patient Microglia Exhibit Enhanced Aging and Unique Transcriptional Activation , 2020, Cell reports.

[7]  J. Salazar,et al.  Microglial Activation in the Retina of a Triple-Transgenic Alzheimer’s Disease Mouse Model (3xTg-AD) , 2020, International journal of molecular sciences.

[8]  L. Souza,et al.  Inflammatory and Pro-resolving Mediators in Frontotemporal Dementia and Alzheimer’s Disease , 2019, Neuroscience.

[9]  B. Lamb,et al.  Inflammation as a central mechanism in Alzheimer's disease , 2018, Alzheimer's & dementia.

[10]  H. Morrison,et al.  Quantifying Microglia Morphology from Photomicrographs of Immunohistochemistry Prepared Tissue Using ImageJ , 2018, Journal of visualized experiments : JoVE.

[11]  M. Viitanen,et al.  PSEN1 Mutant iPSC-Derived Model Reveals Severe Astrocyte Pathology in Alzheimer's Disease , 2017, Stem cell reports.

[12]  C. Goldsbury,et al.  Microglia show altered morphology and reduced arborization in human brain during aging and Alzheimer's disease , 2017, Brain pathology.

[13]  B. Barres,et al.  Reactive Astrocytes: Production, Function, and Therapeutic Potential. , 2017, Immunity.

[14]  M. Colonna,et al.  Microglia Function in the Central Nervous System During Health and Neurodegeneration. , 2017, Annual review of immunology.

[15]  Michael D. Cahalan,et al.  iPSC-Derived Human Microglia-like Cells to Study Neurological Diseases , 2017, Neuron.

[16]  A. Verkhratsky,et al.  Aberrant iPSC-derived human astrocytes in Alzheimer's disease , 2017, Cell Death & Disease.

[17]  T. Wisniewski,et al.  Alzheimer’s disease: experimental models and reality , 2017, Acta Neuropathologica.

[18]  Madeline A. Lancaster,et al.  Human cerebral organoids recapitulate gene expression programs of fetal neocortex development , 2015, Proceedings of the National Academy of Sciences.

[19]  J. de Magalhães,et al.  A comparison of human and mouse gene co-expression networks reveals conservation and divergence at the tissue, pathway and disease levels , 2015, BMC Evolutionary Biology.

[20]  Burkhard Becher,et al.  Immune attack: the role of inflammation in Alzheimer disease , 2015, Nature Reviews Neuroscience.

[21]  B. Giros,et al.  Morphometric characterization of microglial phenotypes in human cerebral cortex , 2014, Journal of Neuroinflammation.

[22]  Jialin C. Zheng,et al.  Characterization of Induced Neural Progenitors from Skin Fibroblasts by a Novel Combination of Defined Factors , 2013, Scientific Reports.

[23]  Huaxi Xu,et al.  Apolipoprotein E and Alzheimer disease: risk, mechanisms and therapy , 2013, Nature Reviews Neurology.

[24]  F. Saravia,et al.  Environmental enrichment prevents astroglial pathological changes in the hippocampus of APP transgenic mice, model of Alzheimer's disease , 2013, Experimental Neurology.

[25]  J. Rutledge,et al.  Postprandial apoE Isoform and Conformational Changes Associated with VLDL Lipolysis Products Modulate Monocyte Inflammation , 2012, PloS one.

[26]  T. Pufe,et al.  Functional and physical interactions between formyl‐peptide‐receptors and scavenger receptor MARCO and their involvement in amyloid beta 1–42‐induced signal transduction in glial cells , 2010, Journal of neurochemistry.

[27]  C. Brayne,et al.  Astrocyte phenotype in relation to Alzheimer-type pathology in the ageing brain , 2010, Neurobiology of Aging.

[28]  Lars Bertram,et al.  Genome-wide association studies in Alzheimer's disease. , 2009, Human molecular genetics.

[29]  E. Londos,et al.  Soluble adhesion molecules and angiotensin-converting enzyme in dementia , 2007, Neurobiology of Disease.

[30]  J. Pober,et al.  Increased ICAM-1 expression causes endothelial cell leakiness, cytoskeletal reorganization and junctional alterations. , 2007, The Journal of investigative dermatology.

[31]  R. Tanzi,et al.  Twenty Years of the Alzheimer’s Disease Amyloid Hypothesis: A Genetic Perspective , 2005, Cell.

[32]  M. Michalopoulou,et al.  The role of soluble intercellular adhesion molecules in neurodegenerative disorders , 2005, Journal of the Neurological Sciences.

[33]  W. Markesbery,et al.  Associations of cortical astrogliosis with cognitive performance and dementia status. , 2005, Journal of Alzheimer's disease : JAD.

[34]  C. Kawas,et al.  Immune reactive cells in senile plaques and cognitive decline in Alzheimer’s disease , 2003, Neurobiology of Aging.

[35]  A. Roses,et al.  Human apoE3 but not apoE4 rescues impaired astrocyte activation in apoE null mice , 2003, Neurobiology of Disease.

[36]  Roger N Gunn,et al.  In-vivo measurement of activated microglia in dementia , 2001, The Lancet.

[37]  D. Selkoe.  Alzheimer's disease: genes, proteins, and therapy. , 2001, Physiological reviews.

[38]  Johnm . Taylor,et al.  Two Distal Downstream Enhancers Direct Expression of the Human Apolipoprotein E Gene to Astrocytes in the Brain , 2001, The Journal of Neuroscience.

[39]  M. Prinz,et al.  Murine Microglial Cells Produce and Respond to Interleukin‐18 , 1999, Journal of neurochemistry.

[40]  Y. Fukuuchi,et al.  Microglia-specific localisation of a novel calcium binding protein, Iba1. , 1998, Brain research. Molecular brain research.

[41]  Y. Ihara,et al.  Astrocytes containing amyloid beta-protein (Abeta)-positive granules are associated with Abeta40-positive diffuse plaques in the aged human brain. , 1998, The American journal of pathology.

[42]  Shuxian Hu,et al.  Cytokine regulation of human microglial cell IL-8 production. , 1998, Journal of immunology.

[43]  M. Gearing,et al.  Astrocyte‐Apolipoprotein E Associations in Senile Plaques in Alzheimer Disease and Vascular Lesions: A Regional Immunohistochemical Study , 1997, Journal of neuropathology and experimental neurology.

[44]  J. Haines,et al.  Gene dose of apolipoprotein E type 4 allele and the risk of Alzheimer's disease in late onset families. , 1993, Science.

[45]  C. Cotman,et al.  Trophic effects of interleukin-4, -7 and -8 on hippocampal neuronal cultures: potential involvement of glial-derived factors , 1993, Brain Research.

[46]  W. Griffin,et al.  Brain interleukin 1 and S-100 immunoreactivity are elevated in Down syndrome and Alzheimer disease. , 1989, Proceedings of the National Academy of Sciences of the United States of America.

[47]  P. Eikelenboom,et al.  Microglial cells around amyloid plaques in Alzheimer's disease express leucocyte adhesion molecules of the LFA-1 family , 1989, Neuroscience Letters.

[48]  Mei Xu,et al.  Pathological Changes in Alzheimer's Disease Analyzed Using Induced Pluripotent Stem Cell-Derived Human Microglia-Like Cells. , 2019, Journal of Alzheimer's disease : JAD.

[49]  T. Montine,et al.  Apolipoprotein E-specific innate immune response in astrocytes from targeted replacement mice , 2006 .

[50]  I. Tooyama,et al.  Expression of intercellular adhesion molecule (ICAM)-1 by a subset of astrocytes in Alzheimer disease and some other degenerative neurological disorders , 2004, Acta Neuropathologica.

[51]  B. de Strooper,et al.  Pathogenic APP mutations near the gamma-secretase cleavage site differentially affect Abeta secretion and APP C-terminal fragment stability. , 2001, Human molecular genetics.

[52]  T. G. Smith,et al.  Surface complexity of human neocortical astrocytic cells: changes with development, aging, and dementia. , 1995, Journal fur Hirnforschung.

[53]  P. Wesseling,et al.  Accumulation of intercellular adhesion molecule-1 in senile plaques in brain tissue of patients with Alzheimer's disease. , 1994, The American journal of pathology.

[54]  D. Marshak,et al.  Increased S100 beta neurotrophic activity in Alzheimer's disease temporal lobe. , 1992, Neurobiology of aging.