| 1. |
- Azevedo, Carla, et al.
(författare)
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Generation of an induced pluripotent stem cell line (CSC-32) from a patient with Parkinson's disease carrying a heterozygous variation p.A53T in the SNCA gene
- 2020
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Ingår i: Stem Cell Research. - : Elsevier BV. - 1873-5061. ; 43
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Tidskriftsartikel (refereegranskat)abstract
- Here, we describe the generation of an induced pluripotent stem cell (iPSC) line, from a male patient diagnosed with Parkinson's disease (PD). The patient carries a heterozygous variation p.A53T in the SNCA gene. Skin fibroblasts were reprogrammed using the non-integrating Sendai virus technology to deliver OCT3/4, SOX2, c-MYC and KLF4 factors. The generated iPSC line (CSC-32) preserved the mutation, displayed expression of common pluripotency markers, differentiated into derivatives of the three germ layers, and exhibited a normal karyotype. The clone CSC-32B is presented thereafter; it can be used to study the mechanisms underlying PD pathogenesis.
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| 2. |
- Azevedo, Carla
(författare)
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Oligodendroglial (dys)function in alpha-synucleinopathies
- 2020
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Doktorsavhandling (övrigt vetenskapligt/konstnärligt)abstract
- The complexity of a-synucleinopathies, which include multiple system atrophy (MSA) and Parkinson’s disease (PD), is not entirely understood. It is known that pathological accumulation of a-synuclein (a-syn) into proteinaceous aggregates is a cellular hallmark of these diseases. a-Syn positive aggregates appear in neurons in PD and dementia with Lewy bodies (DLB), and in oligodendrocytes in MSA. Much research has focused on using animal models that pathocopy and phenocopy PD and MSA to investigate the disease pathogenesis. However, as no genetic link has been associated with MSA, it is possible that current transgenic models do not fully reflect the human pathology. The discovery of induced pluripotent stem cells (iPSCs) changed life science, and allowed for the first time to conduct large scale experimental work using patient cells. Since these cells are embryonic pluripotentlike, their use can help to unravel early disease mechanisms, as they can be differentiated into young brain cells.Despite intense research efforts made to understand the pathogenesis of PD and MSA, several questions remain unanswered, in particular the origin of a-syn in oligodendrocytes and possible toxicity to these cells.The work presented in this thesis aims to 1) generate patient iPSC- based models, 2) develop efficient protocols to generate dopaminergic neurons and oligodendrocytes from iPSCs, and 3) study the role of oligodendrocytes in synucleinopathies, to gain insights into oligodendrocyte (dys)function in PD and MSA. Here, we report that duringoligodendrocyte development and in the human brain, the SNCA gene encoding for a-syn is differentially expressed in oligodendrocytes. Since neurons expressing pathogenic forms of a-syn exhibit cellular alterations, we hypothesized that oligodendrocytes having the same genetic background should also be affected.For the first time, we show that iPSC-derived oligodendrocytes generated from PD patients carrying the variation p.A53T in a-syn or a triplication of the SNCA locus, and from MSA patients, display impaired differentiation and maturation. This was further supported by observations from experiments involving mouse embryonic stem cell(mESC)-derived oligodendrocytes generated from the M83 transgenic mouse model of PD. Moreover, we demonstrate that p.A53T a-syn and MSA oligodendrocytes exhibit a deviation in their phenotype, adopting an immune-reactive phenotype and not myelinating oligodendroglia. Finally, our transcriptomic data further revealalterations in innate inflammatory components, with differential expression of complement proteins, MHC-class and immune-proteasome genes in p.A53T a-syn and MSA oligodendrocytes. This thesis is composed of a unique set of studies addressing crucial questions related to the origin of a-syn in oligodendroglia, and focuses on elucidating the cellular alterations in oligodendrocytes in PD and MSA.
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| 3. |
- Azevedo, Carla, et al.
(författare)
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Parkinsons disease and multiple system atrophy patient iPSC-derived oligodendrocytes exhibit alpha-synuclein-induced changes in maturation and immune reactive properties
- 2022
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Ingår i: Proceedings of the National Academy of Sciences of the United States of America. - : National Academy of Sciences. - 0027-8424 .- 1091-6490. ; 119:12
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Tidskriftsartikel (refereegranskat)abstract
- Limited evidence has shed light on how aSYN proteins affect the oligodendrocyte phenotype and pathogenesis in synucleinopathies that include Parkinsons disease (PD) and multiple system atrophy (MSA). Here, we investigated early transcriptomic changes within PD and MSA O4(+) oligodendrocyte lineage cells (OLCs) generated from patient-induced pluripotent stem cells (iPSCs). We found impaired maturation of PD and MSA O4(+) OLCs compared to controls. This phenotype was associated with changes in the human leukocyte antigen (HLA) genes, the immunoproteasome subunit PSMB9, and the complement component C4b for aSYN p.A53T and MSA O4(+) OLCs, but not in SNCA(trip) O4(+) OLCs despite high levels of aSYN assembly formation. Moreover, SNCA overexpression resulted in the development of O4(+) OLCs, whereas exogenous treatment with aSYN species led to significant toxicity. Notably, transcriptome profiling of genes encoding proteins forming Lewy bodies and glial cytoplasmic inclusions revealed clustering of PD aSYN p.A53T O4(+) OLCs with MSA O4(+) OLCs. Our work identifies early phenotypic and pathogenic changes within human PD and MSA O4(+) OLCs.
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| 4. |
- Bogetofte, Helle, et al.
(författare)
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PARK2 mutation causes metabolic disturbances and impaired survival of human iPSC-derived neurons
- 2019
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Ingår i: Frontiers in Cellular Neuroscience. - : Frontiers Media SA. - 1662-5102. ; 13
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Tidskriftsartikel (refereegranskat)abstract
- The protein parkin, encoded by the PARK2 gene, is vital for mitochondrial homeostasis, and although it has been implicated in Parkinson’s disease (PD), the disease mechanisms remain unclear. We have applied mass spectrometry-based proteomics to investigate the effects of parkin dysfunction on the mitochondrial proteome in human isogenic induced pluripotent stem cell-derived neurons with and without PARK2 knockout (KO). The proteomic analysis quantified nearly 60% of all mitochondrial proteins, 119 of which were dysregulated in neurons with PARK2 KO. The protein changes indicated disturbances in oxidative stress defense, mitochondrial respiration and morphology, cell cycle control, and cell viability. Structural and functional analyses revealed an increase in mitochondrial area and the presence of elongated mitochondria as well as impaired glycolysis and lactate-supported respiration, leading to an impaired cell survival in PARK2 KO neurons. This adds valuable insight into the effect of parkin dysfunction in human neurons and provides knowledge of disease-related pathways that can potentially be targeted for therapeutic intervention.
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| 5. |
- Chari, Arpita, 1986, et al.
(författare)
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Dynamic capabilities for circular manufacturing supply chains-Exploring the role of Industry 4.0 and resilience
- 2022
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Ingår i: Business Strategy and the Environment. - : Wiley. - 1099-0836 .- 0964-4733. ; 31:5, s. 2500-2517
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Tidskriftsartikel (refereegranskat)abstract
- An organisation's sustainability performance is influenced by its capabilities (skills, resources and competences) which in turn affects the performance of its entire supply chain. However, recent research has not sufficiently explored the convergence of dynamic capabilities, circular economy, resilience and Industry 4.0 concepts for manufacturing supply chains. Therefore, this study aims to identify how dynamic capabilities theory can enable circular and resilient supply chains. A qualitative research process was deployed in three stages: literature review, European project and nine expert interviews. Key investigative variables were used to identify capabilities used in manufacturing, and five research propositions were developed to address the gaps found in literature. The empirical data helped reveal challenges to circular economy implementation and validate the literature findings. The main contributions include a dynamic capabilities model, a causal relationship model and five research propositions for circular economy implementation.
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| 6. |
- Chumarina, Margarita, et al.
(författare)
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Cellular alterations identified in pluripotent stem cell-derived midbrain spheroids generated from a female patient with progressive external ophthalmoplegia and parkinsonism who carries a novel variation (p.Q811R) in the POLG1 gene
- 2019
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Ingår i: Acta Neuropathologica Communications. - : Springer Science and Business Media LLC. - 2051-5960. ; 7:1
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Tidskriftsartikel (refereegranskat)abstract
- Variations in the POLG1 gene encoding the catalytic subunit of the mitochondrial DNA polymerase gamma, have recently been associated with Parkinson's disease (PD), especially in patients diagnosed with progressive external ophthalmoplegia (PEO). However, the majority of the studies reporting this association mainly focused on the genetic identification of the variation in POLG1 in PD patient primary cells, and determination of mitochondrial DNA copy number, providing little information about the cellular alterations existing in patient brain cells, in particular dopaminergic neurons. Therefore, through the use of induced pluripotent stem cells (iPSCs), we assessed cellular alterations in novel p.Q811R POLG1 (POLG1Q811R) variant midbrain dopaminergic neuron-containing spheroids (MDNS) from a female patient who developed early-onset PD, and compared them to cultures derived from a healthy control of the same gender. Both POLG1 variant and control MDNS contained functional midbrain regionalized TH/FOXA2-positive dopaminergic neurons, capable of releasing dopamine. Western blot analysis identified the presence of high molecular weight oligomeric alpha-synuclein in POLG1Q811R MDNS compared to control cultures. In order to assess POLG1Q811R-related cellular alterations within the MDNS, we applied mass-spectrometry based quantitative proteomic analysis. In total, 6749 proteins were identified, with 61 significantly differentially expressed between POLG1Q811R and control samples. Pro-And anti-inflammatory signaling and pathways involved in energy metabolism were altered. Notably, increased glycolysis in POLG1Q811R MDNS was suggested by the increase in PFKM and LDHA levels and confirmed using functional analysis of glycolytic rate and oxygen consumption levels. Our results validate the use of iPSCs to assess cellular alterations in relation to PD pathogenesis, in a unique PD patient carrying a novel p.Q811R variation in POLG1, and identify several altered pathways that may be relevant to PD pathogenesis.
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| 7. |
- Chumarina, Margarita, et al.
(författare)
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Derivation of mouse embryonic stem cell lines from tyrosine hydroxylase reporter mice crossed with a human SNCA transgenic mouse model of Parkinson's disease
- 2017
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Ingår i: Stem Cell Research. - : Elsevier BV. - 1876-7753 .- 1873-5061. ; 19, s. 17-20
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Tidskriftsartikel (refereegranskat)abstract
- Mouse embryonic stem cell (mESC) lines were derived by crossing heterozygous transgenic (tg) mice expressing green fluorescent protein (GFP) under the control of the rat tyrosine hydroxylase (TH) promoter, with homozygous alpha-synuclein (aSYN) mice expressing human mutant SNCAA53T under the control of the mouse Prion promoter (MoPrP), or wildtype (WT) mice. The expression of GFP and human aSYN was validated by immunocytochemistry in midbrain neuron cultures upon differentiation of mESC lines using stromal cell-derived inducing activity. These mESC lines can help to study the impact of human aSYN expression in neurons and oligodendrocytes, and also trace GFP-expressing midbrain neurons.
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| 8. |
- Djelloul, Mehdi, et al.
(författare)
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Alpha-Synuclein Expression in the Oligodendrocyte Lineage: an In Vitro and In Vivo Study Using Rodent and Human Models.
- 2015
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Ingår i: Stem Cell Reports. - : Elsevier BV. - 2213-6711. ; 5:2, s. 174-184
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Tidskriftsartikel (refereegranskat)abstract
- In this study, we sought evidence for alpha-synuclein (ASYN) expression in oligodendrocytes, as a possible endogenous source of ASYN to explain its presence in glial inclusions found in multiple system atrophy (MSA) and Parkinson's disease (PD). We identified ASYN in oligodendrocyte lineage progenitors isolated from the rodent brain, in oligodendrocytes generated from embryonic stem cells, and in induced pluripotent stem cells produced from fibroblasts of a healthy individual and patients diagnosed with MSA or PD, in cultures in vitro. Notably, we observed a significant decrease in ΑSYN during oligodendrocyte maturation. Additionally, we show the presence of transcripts in PDGFRΑ/CD140a(+) cells and SOX10(+) oligodendrocyte lineage nuclei isolated by FACS from rodent and human healthy and diseased brains, respectively. Our work identifies ASYN in oligodendrocyte lineage cells, and it offers additional in vitro cellular models that should provide significant insights of the functional implication of ASYN during oligodendrocyte development and disease.
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| 9. |
- Djelloul, Mehdi, et al.
(författare)
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Reporting on methods to generate and purify rodent and human oligodendrocytes from different sources
- 2017
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Ingår i: Stem Cell Research. - : Elsevier BV. - 1873-5061. ; 20, s. 58-66
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Tidskriftsartikel (refereegranskat)abstract
- Oligodendrocytes are part of the glial cells located in the central nervous system, capable of providing trophic support to neurons and ensheathing their axons. These cells can become dysfunctional under pathologic condition. Rodent and human pluripotent stem cells are inexhaustible sources for producing oligodendrocytes that can be used for disease modeling and cell replacement therapy studies. They also offer many opportunities to model the contribution of oligodendrocytes in non-genetic disorders such as multiple system atrophy. In this method article, we provide robust and reproducible differentiation protocols to obtain oligodendrocyte progenitor cells and purify them using fluorescence activated cell sorting.
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| 10. |
- Gustavsson, Nadja, et al.
(författare)
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Generation of an induced pluripotent stem cell line (CSC-46) from a patient with Parkinson's disease carrying a novel p.R301C mutation in the GBA gene
- 2019
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Ingår i: Stem Cell Research. - : Elsevier BV. - 1873-5061. ; 34
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Tidskriftsartikel (refereegranskat)abstract
- Mutations in the glucocerebrosidase (GBA) gene have been associated with the development of Parkinson's disease (PD). An induced pluripotent stem cell (iPSC) line was generated from a 60-year old patient diagnosed with PD and carrying a new mutation variant p.R301C in GBA. Using non-integrating Sendai virus-based technology, we utilized OCT3/4, SOX2, c-MYC and KLF4 transcription factors to reprogram skin fibroblasts into iPSCs. The generated iPSC line retained the mutation, displayed expression of common pluripotency markers, differentiated into the three germ layers, and exhibited normal karyotype. The iPSC line can be further used for studying PD pathogenesis.
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