his.sePublications
Change search
Link to record
Permanent link

Direct link
BETA
Publications (7 of 7) Show all publications
Delsing, L., Dönnes, P., Sánchez, J., Clausen, M., Voulgaris, D., Falk, A., . . . Synnergren, J. (2018). Barrier properties and transcriptome expression in human iPSC-derived models of the blood-brain barrier. Stem Cells, 36(12), 1816-1827
Open this publication in new window or tab >>Barrier properties and transcriptome expression in human iPSC-derived models of the blood-brain barrier
Show others...
2018 (English)In: Stem Cells, ISSN 1066-5099, E-ISSN 1549-4918, Vol. 36, no 12, p. 1816-1827Article in journal (Refereed) Published
Abstract [en]

Cell-based models of the blood-brain barrier (BBB) are important for increasing the knowledge of BBB formation, degradation and brain exposure of drug substances. Human models are preferred over animal models because of inter-species differences in BBB structure and function. However, access to human primary BBB tissue is limited and has shown degeneration of BBB functions in vitro. Human induced pluripotent stem cells (iPSCs) can be used to generate relevant cell types to model the BBB with human tissue. We generated a human iPSC-derived model of the BBB that includes endothelial cells in co-culture with pericytes, astrocytes and neurons. Evaluation of barrier properties showed that the endothelial cells in our co-culture model have high transendothelial electrical resistance, functional efflux and ability to discriminate between CNS permeable and non-permeable substances. Whole genome expression profiling revealed transcriptional changes that occur in co-culture, including upregulation of tight junction proteins such as claudins and neurotransmitter transporters. Pathway analysis implicated changes in the WNT, TNF and PI3K-Akt pathways upon co-culture. Our data suggests that co-culture of iPSC-derived endothelial cells promotes barrier formation on a functional and transcriptional level. The information about gene expression changes in co-culture can be used to further improve iPSC-derived BBB models through selective pathway manipulation.

Keywords
blood–brain barrier, co-culture, hiPSC, in vitro models, transcriptome, endothelial cells
National Category
Medical Biotechnology (with a focus on Cell Biology (including Stem Cell Biology), Molecular Biology, Microbiology, Biochemistry or Biopharmacy)
Research subject
Bioinformatics; INF502 Biomarkers
Identifiers
urn:nbn:se:his:diva-16283 (URN)10.1002/stem.2908 (DOI)000455838500004 ()30171748 (PubMedID)2-s2.0-85056115357 (Scopus ID)
Available from: 2018-10-08 Created: 2018-10-08 Last updated: 2019-02-15Bibliographically approved
Lundin, A., Delsing, L., Clausen, M., Ricchiuto, P., Sanchez, J., Sabirsh, A., . . . Falk, A. (2018). Human iPS-Derived Astroglia from a Stable Neural Precursor State Show Improved Functionality Compared with Conventional Astrocytic Models. Stem Cell Reports, 10(3), 1030-1045
Open this publication in new window or tab >>Human iPS-Derived Astroglia from a Stable Neural Precursor State Show Improved Functionality Compared with Conventional Astrocytic Models
Show others...
2018 (English)In: Stem Cell Reports, ISSN 2213-6711, Vol. 10, no 3, p. 1030-1045Article in journal (Refereed) Published
Abstract [en]

In vivo studies of human brain cellular function face challenging ethical and practical difficulties. Animal models are typically used but display distinct cellular differences. One specific example is astrocytes, recently recognized for contribution to neurological diseases and a link to the genetic risk factor apolipoprotein E (APOE). Current astrocytic in vitro models are questioned for lack of biological characterization. Here, we report human induced pluripotent stem cell (hiPSC)-derived astroglia (NES-Astro) developed under defined conditions through long-term neuroepithelial-like stem (ltNES) cells. We characterized NES-Astro and astrocytic models from primary sources, astrocytoma (CCF-STTG1), and hiPSCs through transcriptomics, proteomics, glutamate uptake, inflammatory competence, calcium signaling response, and APOE secretion. Finally, we assess modulation of astrocyte biology using APOE-annotated compounds, confirming hits of the cholesterol biosynthesis pathway in adult and hiPSC-derived astrocytes. Our data show large diversity among astrocytic models and emphasize a cellular context when studying astrocyte biology.

Keywords
apolipoproteins E, astrocytes, calcium signaling, cell differentiation, drug discovery, glutamate plasma membrane transport proteins, high-throughput screening assays, induced pluripotent stem cells, neurodegenerative diseases, neuroinflammation
National Category
Cell Biology
Research subject
Bioinformatics; INF502 Biomarkers
Identifiers
urn:nbn:se:his:diva-14757 (URN)10.1016/j.stemcr.2018.01.021 (DOI)000427349300028 ()29456185 (PubMedID)2-s2.0-85042043028 (Scopus ID)
Available from: 2018-02-20 Created: 2018-02-20 Last updated: 2019-02-14Bibliographically approved
Kristensson, L., Lundin, A., Gustafsson, D., Fryklund, J., Fex, T., Delsing, L. & Ryberg, E. (2018). Plasminogen binding inhibitors demonstrate unwanted activities on GABAA and glycine receptors in human iPSC derived neurons. Neuroscience Letters, 681, 37-43, Article ID S0304-3940(18)30351-3.
Open this publication in new window or tab >>Plasminogen binding inhibitors demonstrate unwanted activities on GABAA and glycine receptors in human iPSC derived neurons
Show others...
2018 (English)In: Neuroscience Letters, ISSN 0304-3940, E-ISSN 1872-7972, Vol. 681, p. 37-43, article id S0304-3940(18)30351-3Article in journal (Refereed) Published
Abstract [en]

Plasminogen binding inhibitors (PBIs) reduce the risk of bleeding in hemorrhagic conditions. However, generic PBIs are also associated with an increased risk of seizures, an adverse effect linked to unwanted activities towards inhibitory neuronal receptors. Development of novel PBIs serve to remove compounds with such properties, but progress is limited by a lack of higher throughput methods with human translatability. Herein we apply human induced pluripotent stem cell (hiPSC) derived neurons in combination with dynamic mass redistribution (DMR) technology to demonstrate robust and reproducible modulation of both GABAA and glycine receptors. These cells respond to GABA (EC50 0.33 ± 0.18 μM), glycine (EC50 11.0 ± 3.7 μM) and additional ligands in line with previous reports from patch clamp technologies. Additionally, we identify and characterize a competitive antagonistic behavior of the prototype inhibitor and drug tranexamic acid (TXA). Finally, we demonstrate proof of concept for effective counter-screening of lead series compounds towards unwanted GABAAreceptor activities. No activity was observed for a previously identified PBI candidate drug, AZD6564, whereas a discontinued analog, AZ13267257, could be characterized as a potent GABAA receptor agonist.

Keywords
Drug screening, Dynamic mass redistribution, GABA(A) receptor, Glycine receptor, Pharmacology, hiPSC derived neurons
National Category
Cell Biology
Research subject
Bioinformatics; INF502 Biomarkers
Identifiers
urn:nbn:se:his:diva-15263 (URN)10.1016/j.neulet.2018.05.018 (DOI)000442191200008 ()29758302 (PubMedID)2-s2.0-85047397711 (Scopus ID)
Available from: 2018-06-04 Created: 2018-06-04 Last updated: 2019-02-14Bibliographically approved
Skillbäck, T., Delsing, L., Synnergren, J., Mattsson, N., Janelidze, S., Nägga, K., . . . Zetterberg, H. (2017). CSF/serum albumin ratio in dementias: a cross-sectional study on 1861 patients. Neurobiology of Aging, 59, 1-9
Open this publication in new window or tab >>CSF/serum albumin ratio in dementias: a cross-sectional study on 1861 patients
Show others...
2017 (English)In: Neurobiology of Aging, ISSN 0197-4580, E-ISSN 1558-1497, Vol. 59, p. 1-9Article in journal (Refereed) Published
Abstract [en]

A connection between dementias and blood-brain barrier (BBB) dysfunction has been suggested, but previous studies have yielded conflicting results. We examined cerebrospinal fluid (CSF)/serum albumin ratio in a large cohort of patients diagnosed with Alzheimer's disease (AD, early onset [EAD, n = 130], late onset AD [LAD, n = 666]), vascular dementia (VaD, n = 255), mixed AD and VaD (MIX, n = 362), Lewy body dementia (DLB, n = 50), frontotemporal dementia (FTD, n = 56), Parkinson's disease dementia (PDD, n = 23), other dementias (other, n = 48), and dementia not otherwise specified (NOS, n = 271). We compared CSF/serum albumin ratio to 2 healthy control groups (n = 292, n = 20), between dementia diagnoses, and tested biomarker associations. Patients in DLB, LAD, VaD, MIX, other, and NOS groups had higher CSF/serum albumin ratio than controls. CSF/serum albumin ratio correlated with CSF neurofilament light in LAD, MIX, VaD, and other groups but not with AD biomarkers. Our data show that BBB leakage is common in dementias. The lack of association between CSF/serum albumin ratio and AD biomarkers suggests that BBB dysfunction is not inherent to AD but might represent concomitant cerebrovascular pathology.

Place, publisher, year, edition, pages
Elsevier, 2017
Keywords
Alzheimer's disease, Blood-brain barrier, CSF/serum albumin ratio, Dementia
National Category
Neurology
Research subject
Bioinformatics; INF502 Biomarkers
Identifiers
urn:nbn:se:his:diva-14035 (URN)10.1016/j.neurobiolaging.2017.06.028 (DOI)000411446800001 ()28779628 (PubMedID)2-s2.0-85026625803 (Scopus ID)
Funder
Knowledge Foundation
Available from: 2017-08-24 Created: 2017-08-24 Last updated: 2019-02-14Bibliographically approved
Wobst, H. J., Wesolowski, S. S., Chadchankar, J., Delsing, L., Jacobsen, S., Mukherjee, J., . . . Moss, S. J. (2017). Cytoplasmic Relocalization of TAR DNA-Binding Protein 43 Is Not Sufficient to Reproduce Cellular Pathologies Associated with ALS In vitro. Frontiers in Molecular Neuroscience, 10, Article ID 46.
Open this publication in new window or tab >>Cytoplasmic Relocalization of TAR DNA-Binding Protein 43 Is Not Sufficient to Reproduce Cellular Pathologies Associated with ALS In vitro
Show others...
2017 (English)In: Frontiers in Molecular Neuroscience, ISSN 1662-5099, Vol. 10, article id 46Article in journal (Refereed) Published
Abstract [en]

Mutations in the gene TARDBP, which encodes TAR DNA-binding protein 43 (TDP-43), are a rare cause of familial forms of amyotrophic lateral sclerosis (ALS) and frontotemporal dementia (FTD). While the majority of mutations are found in the C-terminal glycine-rich domain, an alanine to valine amino acid change at position 90 (A90V) in the bipartite nuclear localization signal (NLS) of TDP-43 has been described. This sequence variant has previously been shown to cause cytoplasmic mislocalization of TDP-43 and decrease protein solubility, leading to the formation of insoluble aggregates. Since the A90V mutation has been described both in patients as well as healthy controls, its pathogenic potential in ALS and FTD remains unclear. Here we compare properties of overexpressed A90V to the highly pathogenic M337V mutation. Though both mutations drive mislocalization of the protein to the cytoplasm to the same extent, M337V produces more significant damage in terms of protein solubility, levels of pathogenic phosphorylation, and formation of C-terminal truncated protein species. Furthermore, the M337V, but not the A90V mutant, leads to a downregulation of histone deacetylase 6 and Ras GTPase-activating protein-binding protein. We conclude that in the absence of another genetic or environmental 'hit' the A90V variant is not sufficient to cause the deleterious phenotypes associated with ALS and FTD, despite prominent cytoplasmic protein relocalization of TDP-43.

Keywords
TAR DNA-binding protein 43 (TDP-43), amyotrophic lateral sclerosis (ALS), frontotemporal dementia, mutation, neurodegenerative diseases, protein misfolding disease
National Category
Biochemistry and Molecular Biology
Identifiers
urn:nbn:se:his:diva-14041 (URN)10.3389/fnmol.2017.00046 (DOI)000394648800002 ()28286471 (PubMedID)2-s2.0-85014051932 (Scopus ID)
Available from: 2017-08-25 Created: 2017-08-25 Last updated: 2018-11-23Bibliographically approved
Delsing, L., Synnergren, J., Hicks, R. & Zetterberg, H. (2017). Human iPSC-derived endothelial cells can develop in to brain-like endothelial cells after coculture with primary human brain cells. In: : . Paper presented at International Symposium for Stem Cell Research, Boston, June 14-17, 2017 (pp. 523). International Society for Stem Cell Research
Open this publication in new window or tab >>Human iPSC-derived endothelial cells can develop in to brain-like endothelial cells after coculture with primary human brain cells
2017 (English)Conference paper, Poster (with or without abstract) (Refereed)
Place, publisher, year, edition, pages
International Society for Stem Cell Research, 2017
National Category
Medical Biotechnology (with a focus on Cell Biology (including Stem Cell Biology), Molecular Biology, Microbiology, Biochemistry or Biopharmacy)
Research subject
Bioinformatics
Identifiers
urn:nbn:se:his:diva-14760 (URN)
Conference
International Symposium for Stem Cell Research, Boston, June 14-17, 2017
Available from: 2018-02-20 Created: 2018-02-20 Last updated: 2018-02-26Bibliographically approved
Wobst, H. J., Delsing, L., Brandon, N. J. & Moss, S. J. (2017). Truncation of the TAR DNA-binding protein 43 is not a prerequisite for cytoplasmic relocalization, and is suppressed by caspase inhibition and by introduction of the A90V sequence variant. PLoS ONE, 12(5), Article ID e0177181.
Open this publication in new window or tab >>Truncation of the TAR DNA-binding protein 43 is not a prerequisite for cytoplasmic relocalization, and is suppressed by caspase inhibition and by introduction of the A90V sequence variant
2017 (English)In: PLoS ONE, ISSN 1932-6203, E-ISSN 1932-6203, Vol. 12, no 5, article id e0177181Article in journal (Refereed) Published
Abstract [en]

The RNA-binding and -processing protein TAR DNA-binding protein 43 (TDP-43) is heavily linked to the underlying causes and pathology of neurodegenerative diseases such as amyotrophic lateral sclerosis and frontotemporal lobar degeneration. In these diseases, TDP-43 is mislocalized, hyperphosphorylated, ubiquitinated, aggregated and cleaved. The importance of TDP-43 cleavage in the disease pathogenesis is still poorly understood. Here we detail the use of D-sorbitol as an exogenous stressor that causes TDP-43 cleavage in HeLa cells, resulting in a 35 kDa truncated product that accumulates in the cytoplasm within one hour of treatment. We confirm that the formation of this 35 kDa cleavage product is mediated by the activation of caspases. Inhibition of caspases blocks the cleavage of TDP-43, but does not prevent the accumulation of full-length protein in the cytoplasm. Using D-sorbitol as a stressor and caspase activator, we also demonstrate that the A90V variant of TDP-43, which lies adjacent to the caspase cleavage site within the nuclear localization sequence of TDP-43, confers partial resistance against caspase-mediated generation of the 35 kDa cleavage product.

National Category
Biochemistry and Molecular Biology
Identifiers
urn:nbn:se:his:diva-14040 (URN)10.1371/journal.pone.0177181 (DOI)000401485500016 ()28510586 (PubMedID)2-s2.0-85019583583 (Scopus ID)
Available from: 2017-08-25 Created: 2017-08-25 Last updated: 2018-09-20Bibliographically approved
Organisations
Identifiers
ORCID iD: ORCID iD iconorcid.org/0000-0003-2899-3801

Search in DiVA

Show all publications