Generation of 2 induced pluripotent stem cell lines derived from patients with Parkinson's disease carrying LRRK2 G2385R variant

Cheng, Yu‐Che; Huang, Ching‐Ying; Ho, Ming-Ching; Hsu, Yu-Hung; Syu, Shih-Han; Lu, Huai-En; Lin, Han-I; Lin, Chin‐Hsien; Hsieh, Patrick C.

doi:10.1016/j.scr.2018.01.034

Cited by 8 publications

(4 citation statements)

References 3 publications

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“…This discovery has enabled the study of many diseases as these stem cell lines can be derived from patients suffering from different diseases and disorders. Using patient derived hiPSCs lines and differentiating them into a target tissue type is a powerful way to study diseases, including those affecting the nervous system -like Alzheimer's and Parkinson's (Playne and Connor, 2017;Cheng et al, 2018Cheng et al, , 2019Penney et al, 2019). Often, research on hiPSC models of these diseases is conducted in 2D, despite brain tissue possessing a complex 3D structure.…”

Section: Introductionmentioning

confidence: 99%

3D Bioprinting Pluripotent Stem Cell Derived Neural Tissues Using a Novel Fibrin Bioink Containing Drug Releasing Microspheres

Sharma

Smits

Vega

et al. 2020

Front. Bioeng. Biotechnol.

110

104

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3D bioprinting combines cells with a supportive bioink to fabricate multiscale, multicellular structures that imitate native tissues. Here, we demonstrate how our novel fibrinbased bioink formulation combined with drug releasing microspheres can serve as a tool for bioprinting tissues using human induced pluripotent stem cell (hiPSC)-derived neural progenitor cells (NPCs). Microspheres, small spherical particles that generate controlled drug release, promote hiPSC differentiation into dopaminergic neurons when used to deliver small molecules like guggulsterone. We used the microfluidics based RX1 bioprinter to generate domes with a 1 cm diameter consisting of our novel fibrin-based bioink containing guggulsterone microspheres and hiPSC-derived NPCs. The resulting tissues exhibited over 90% cellular viability 1 day post printing that then increased to 95% 7 days post printing. The bioprinted tissues expressed the early neuronal marker, TUJ1 and the early midbrain marker, Forkhead Box A2 (FOXA2) after 15 days of culture. These bioprinted neural tissues expressed TUJ1 (15 ± 1.3%), the dopamine marker, tyrosine hydroxylase (TH) (8 ± 1%) and other glial markers such as glial fibrillary acidic protein (GFAP) (15 ± 4%) and oligodendrocyte progenitor marker (O4) (4 ± 1%) after 30 days. Also, quantitative polymerase chain reaction (qPCR) analysis showed these bioprinted tissues expressed TUJ1, NURR1 (gene expressed in midbrain dopaminergic neurons), LMX1B, TH, and PAX6 after 30 days. In conclusion, we have demonstrated that using a microsphere-laden bioink to bioprint hiPSC-derived NPCs can promote the differentiation of neural tissue.

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Section: Introductionmentioning

confidence: 99%

3D Bioprinting Pluripotent Stem Cell Derived Neural Tissues Using a Novel Fibrin Bioink Containing Drug Releasing Microspheres

Sharma

Smits

Vega

et al. 2020

Front. Bioeng. Biotechnol.

110

104

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“…The mutation comprises six pathogenic variants, G2019S, R1441G/C/H, I1371V, , R1628P, , G2385R, and I2020T, showing varied prevalences based on ethnicity. The kinase domain mutation is predominant in Caucasian PD patients; however, the prevalence of the pathogenic GTPase domain mutation has been reported in Indian and East-Asian populations and is estimated to increase the risk of PD by about twofold. − …”

Section: Introductionmentioning

confidence: 99%

Dopaminergic Neurons Differentiated from LRRK2 I1371V-Induced Pluripotent Stem Cells Display a Lower Yield, α-Synuclein Pathology, and Functional Impairment

Jagtap

Potdar

Yadav

et al. 2022

ACS Chem. Neurosci.

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Being a large multidomain protein, LRRK2 has several confirmed pathological mutant variants for PD, and the incidence of these variants shows ethnicity biases. I1371V, a mutation in the GTPase domain, has been reported in East-Asian populations, but there are no studies reported on dopaminergic (DA) neurons differentiated from this variant. The aim here was to assess the yield, function, and α-synuclein pathology of DA neurons differentiated from LRRK2 I1371V iPSCs. FACS analysis of neural progenitors (NPs) showed a comparable immunopositive population of cells for neural and glial progenitor markers nestin and S100β; however, NPs from I1371V iPSCs showed lower clonogenic and proliferative capacities than healthy control NPs as determined by the neurosphere assay and Ki67 expression. Floor plate cells obtained from I1371V NPs primed with FGF8 showed distinctly lower immunopositivity for FOXA2 and CLIC5 than healthy control FPCs and similar DOC2B expression. On SHH addition, a similar mature neuronal population was obtained from both groups; however, the yield of TH-immunopositive cells was significantly lower in I1371V, with lower expression of mature DA neuronal markers En1, Nurr1, and DAT. Vesicular dopamine release and intracellular Ca2+ response with KCl stimulation were lower in I1371V DA neurons, along with a significantly reduced expression of resting vesicle marker VMAT2. A concurrently lower expression of PSD95/Syn-I immunopositive puncta was observed in I1371V differentiated cells. Further, higher phosphorylation of α-synuclein and aggregation of oligomeric α-synuclein in I1371V DA neurons were observed. Our data demonstrated conclusively for the first time that mutations in the I1371V allele of LRRK2 showed developmental deficit from the FPC stage and generated a lower yield/number of TH-immunopositive neurons with impairment in their function and synapse density along with increased α-synuclein pathology.

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“…The LRRK2 G2019S mutation has the effect of enhancing LRRK2 kinase activity, and the first LRRK2 iPSCs were established in 2012 [55]. Hereafter, LRRK2 iPSC models carrying mutations G2385R [56], R1628P [57], N551K [58], and S1647T [59] were also established. The role of LRRK2 in iPSC-derived neurons is depicted in Figure 2.…”

Section: Leucine-rich Repeat Kinase 2 (Lrrk2)mentioning

confidence: 99%

Modeling Parkinson’s Disease Using Induced Pluripotent Stem Cells

Mao

Fan

et al. 2020

Stem Cells International

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Parkinson’s disease (PD) is the second most common neurodegenerative disease. The molecular mechanisms of PD at the cellular level involve oxidative stress, mitochondrial dysfunction, autophagy, axonal transport, and neuroinflammation. Induced pluripotent stem cells (iPSCs) with patient-specific genetic background are capable of directed differentiation into dopaminergic neurons. Cell models based on iPSCs are powerful tools for studying the molecular mechanisms of PD. The iPSCs used for PD studies were mainly from patients carrying mutations in synuclein alpha (SNCA), leucine-rich repeat kinase 2 (LRRK2), PTEN-induced putative kinase 1 (PINK1), parkin RBR E3 ubiquitin protein ligase (PARK2), cytoplasmic protein sorting 35 (VPS35), and variants in glucosidase beta acid (GBA). In this review, we summarized the advances in molecular mechanisms of Parkinson’s disease using iPSC models.

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Generation of 2 induced pluripotent stem cell lines derived from patients with Parkinson's disease carrying LRRK2 G2385R variant

Cited by 8 publications

References 3 publications

3D Bioprinting Pluripotent Stem Cell Derived Neural Tissues Using a Novel Fibrin Bioink Containing Drug Releasing Microspheres

3D Bioprinting Pluripotent Stem Cell Derived Neural Tissues Using a Novel Fibrin Bioink Containing Drug Releasing Microspheres

Dopaminergic Neurons Differentiated from LRRK2 I1371V-Induced Pluripotent Stem Cells Display a Lower Yield, α-Synuclein Pathology, and Functional Impairment

Modeling Parkinson’s Disease Using Induced Pluripotent Stem Cells

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