Generation and characterization of a human induced pluripotent stem cell line from a familial Alzheimer's disease PSEN1 T119I patient

ISAJA, LUCIANA; RODRÍGUEZ VARELA, MARÍA SOLEDAD; MARAZITA, MARIELA; MUCCI, SOFÍA; ITZCOVICH, TATIANA; CHEM-MENDEZ, PATRICIO; NIIKADO, MATIAS; ALLEGRI, RICARDO; MARTINETTO, HORACIO; SEVLEVER, GUSTAVO EMILIO; SCASSA, MARÍA ELIDA; SURACE, EZEQUIEL IGNACIO; ROMORINI, LEONARDO

Buenos Aires

Congreso; Reunión de Sociedades Biociencias 2020, SAIC.SAI.SAFIS; 2020

SAIC SAI SAFIS

Alzheimer’s disease (AD) is a neurodegenerative proteinopathy which is the main cause of dementia in adults. It is estimated that 5% of cases are caused by inherited mutations in genes such as Pre- senilin-1 (PSEN1). Previously, our group reported a novel heterozy- gous variant in PSEN1 (c.356C>T; p.T119I) in an Argentine family with early- and late-onset AD. In order to functionally validate this variant, we aimed to generate a human induced pluripotent stem cell (hiPSC) line from dermic fibroblasts (DF) of a male mutation carrier. To this purpose, we first infected patient DF with a STEMCCA lenti- viral vector encoding the Yamanaka reprogramming factors (OCT-4, KLF4, SOX2 y c-MYC) and obtained two clones, termed FFAD1.2c4 and FFAD1.2c8. Both clones exhibited typical hiPSCs morphologi- cal characteristics (formation of compact multicellular colonies with a high nucleus/cytoplasm ratio and distinct colony borders) and high Alkaline Phosphatase (AP) activity. Moreover, we also observed ro- bust expression of different stemness-associated markers (OCT-4, NANOG, SSEA4, TRA1-80, and TRA1-60),) analyzed by immuno- fluorescence and RT-qPCR. Also, using a non-directed method of differentiation (embryoid bodies assay) we demonstrated that at least FFAD1.2c4 had the pluripotent potential to be differentiated into cells from the three germinal layers (mesoderm, endoderm and ectoderm) as judged by RT-qPCR and immunofluorescence anal- ysis of Smooth muscle actin (SMA), Alpha-fetoprotein (AFP) and Nestin differentiation markers expression, respectively. Finally, both clones exhibited the same normal karyotype (46, XY) and PSEN1 T119I genotype as the parental DF cells and silenced ectopic ex- pression of Yamanaka gene. Overall, we successfully obtained an hiPSCs line (FFAD1.2) carrying the AD PSEN1 T119I genotype.