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Gallant, Caroline J.
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Publications (10 of 11) Show all publications
Gallant, C. J. & Landegren, U. (2019). A compendium on single-cell analysis for the curious. The FEBS Journal, 286(8), 1442-1444
Open this publication in new window or tab >>A compendium on single-cell analysis for the curious
2019 (English)In: The FEBS Journal, ISSN 1742-464X, E-ISSN 1742-4658, Vol. 286, no 8, p. 1442-1444Article in journal, Editorial material (Other academic) Published
Place, publisher, year, edition, pages
John Wiley & Sons, 2019
National Category
Biochemistry and Molecular Biology
Identifiers
urn:nbn:se:uu:diva-384089 (URN)10.1111/febs.14820 (DOI)000466783900001 ()31012288 (PubMedID)
Available from: 2019-06-19 Created: 2019-06-19 Last updated: 2019-06-19Bibliographically approved
Batool, T., Fang, J., Jansson, V., Zhao, H., Gallant, C. J., Moustakas, A. & Li, J.-P. (2019). Upregulated BMP-Smad signaling activity in the glucuronyl C5-epimerase knock out MEF cells. Cellular Signalling, 54, 122-129
Open this publication in new window or tab >>Upregulated BMP-Smad signaling activity in the glucuronyl C5-epimerase knock out MEF cells
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2019 (English)In: Cellular Signalling, ISSN 0898-6568, E-ISSN 1873-3913, Vol. 54, p. 122-129Article in journal (Refereed) Published
Abstract [en]

Glucuronyl C5-epimerase (Hsepi) catalyzes the conversion of glucuronic acid to iduronic acid in the process of heparan sulfate biosynthesis. Targeted interruption of the gene, Glce,in mice resulted in neonatal lethality with varied defects in organ development. To understand the molecular mechanisms of the phenotypes, we used mouse embryonic fibroblasts (MEF) as a model to examine selected signaling pathways. Our earlier studies found reduced activities of FGF-2, GDNF, but increased activity of sonic hedgehog in the mutant cells. In this study, we focused on the bone morphogenetic protein (BMP) signaling pathway. Western blotting detected substantially elevated endogenous Smad1/5/8 phosphorylation in the Hsepi mutant (KO) MEF cells, which is reverted by re-expression of the enzyme in the KO cells. The mutant cells displayed an enhanced proliferation and elevated alkaline phosphatase activity, marking higher differentiation, when cultured in osteogenic medium. The high level of Smad1/5/8 phosphorylation was also found in primary calvarial cells isolated from the KO mice. Analysis of the genes involved in the BMP signaling pathway revealed upregulation of a number of BMP ligands, but reduced expression of several Smads and BMP antagonist (Grem1) in the KO MEF cells. The results suggest that Hsepi expression modulates BMP signaling activity, which, at least partially, is associated with defected molecular structure of heparan sulfate expressed in the cells.   

Place, publisher, year, edition, pages
Elsevier, 2019
Keywords
BMP signaling, Heparan sulfate, MEF cells, Smad, Glucuronyl C5-epimerase, Bone Morphogenetic Protein
National Category
Cell and Molecular Biology Medical Biotechnology (with a focus on Cell Biology (including Stem Cell Biology), Molecular Biology, Microbiology, Biochemistry or Biopharmacy)
Identifiers
urn:nbn:se:uu:diva-363254 (URN)10.1016/j.cellsig.2018.11.010 (DOI)000456752900013 ()30458230 (PubMedID)
Funder
Swedish Cancer Society, CAN2015/496Swedish Research Council, 2015-02595Science for Life Laboratory - a national resource center for high-throughput molecular bioscience
Available from: 2018-10-15 Created: 2018-10-15 Last updated: 2019-02-12Bibliographically approved
Larssen, P., Wik, L., Czarnewski, P., Eldh, M., Löf, L., Ronquist, G., . . . Kamali-Moghaddam, M. (2017). Tracing Cellular Origin of Human Exosomes Using Multiplex Proximity Extension Assay. Molecular & cellular proteomics (online), 16(3), 502-511
Open this publication in new window or tab >>Tracing Cellular Origin of Human Exosomes Using Multiplex Proximity Extension Assay
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2017 (English)In: Molecular & cellular proteomics (online), ISSN 1535-9476, E-ISSN 1535-9484, Vol. 16, no 3, p. 502-511Article in journal (Refereed) Published
Abstract [en]

Extracellular vesicles (EVs) are membrane-coated objects such as exosomes and microvesicles, released by many cell-types. Their presence in body fluids and the variable surface composition and content render them attractive potential biomarkers. The ability to determine their cellular origin could greatly move the field forward. We used multiplex proximity extension assays (PEA) to identify with high specificity and sensitivity the protein profiles of exosomes of different origins, including seven cell lines and two different body fluids. By comparing cells and exosomes, we successfully identified the cells originating the exosomes. Furthermore, by principal component analysis of protein patterns human milk EVs and prostasomes released from prostate acinar cells clustered with cell lines from breast and prostate tissues, respectively. Milk exosomes uniquely expressed CXCL5, MIA and KLK6, while prostasomes carried NKX31, GSTP1 and SRC, highlighting that EVs originating from different origins express distinct proteins. In conclusion, PEA provides a powerful protein screening tool in exosome research, for purposes of identifying the cell source of exosomes, or new biomarkers in diseases such as cancer and inflammation.

National Category
Cancer and Oncology
Identifiers
urn:nbn:se:uu:diva-314142 (URN)10.1074/mcp.M116.064725 (DOI)000395670900013 ()28111361 (PubMedID)
Funder
Swedish Research CouncilEU, FP7, Seventh Framework Programme, 259796 294409Swedish Cancer Society, 2013/867Stockholm County Council, 20140405Swedish Heart Lung Foundation, 20140497The Karolinska Institutet's Research FoundationCancer and Allergy Foundation
Available from: 2017-01-28 Created: 2017-01-28 Last updated: 2017-04-20Bibliographically approved
Darmanis, S., Gallant, C. J., Marinescu, V. D., Niklasson, M., Segerman, A., Flamourakis, G., . . . Landegren, U. (2016). Simultaneous Multiplexed Measurement of RNA and Proteins in Single Cells. Cell reports, 14(2), 380-389
Open this publication in new window or tab >>Simultaneous Multiplexed Measurement of RNA and Proteins in Single Cells
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2016 (English)In: Cell reports, ISSN 2211-1247, E-ISSN 2211-1247, Vol. 14, no 2, p. 380-389Article in journal (Refereed) Published
Abstract [en]

Significant advances have been made in methods to analyze genomes and transcriptomes of single cells, but to fully define cell states, proteins must also be accessed as central actors defining a cell's phenotype. Methods currently used to analyze endogenous protein expression in single cells are limited in specificity, throughput, or multiplex capability. Here, we present an approach to simultaneously and specifically interrogate large sets of protein and RNA targets in lysates from individual cells, enabling investigations of cell functions and responses. We applied our method to investigate the effects of BMP4, an experimental therapeutic agent, on early-passage glioblastoma cell cultures. We uncovered significant heterogeneity in responses to treatment at levels of RNA and protein, with a subset of cells reacting in a distinct manner to BMP4. Moreover, we found overall poor correlation between protein and RNA at the level of single cells, with proteins more accurately defining responses to treatment.

National Category
Cell and Molecular Biology
Identifiers
urn:nbn:se:uu:diva-276810 (URN)10.1016/j.celrep.2015.12.021 (DOI)000368101600020 ()26748716 (PubMedID)
Funder
EU, FP7, Seventh Framework Programme, 259796EU, FP7, Seventh Framework Programme, 294409Swedish Research CouncilKnut and Alice Wallenberg FoundationSwedish Cancer Society
Available from: 2016-02-16 Created: 2016-02-16 Last updated: 2018-01-10Bibliographically approved
Namjou, B., Choi, C.-B., Harley, I. T. W., Alarcon-Riquelme, M. E., Kelly, J. A., Glenn, S. B., . . . Bae, S.-C. (2012). Evaluation of TRAF6 in a large multiancestral lupus cohort. Arthritis and Rheumatism, 64(6), 1960-1969
Open this publication in new window or tab >>Evaluation of TRAF6 in a large multiancestral lupus cohort
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2012 (English)In: Arthritis and Rheumatism, ISSN 0004-3591, E-ISSN 1529-0131, Vol. 64, no 6, p. 1960-1969Article in journal (Refereed) Published
Abstract [en]

Objective Systemic lupus erythematosus (SLE) is a heterogeneous autoimmune disease with significant immune system aberrations resulting from complex heritable genetics as well as environmental factors. We undertook to study the role of TRAF6 as a candidate gene for SLE, since it plays a major role in several signaling pathways that are important for immunity and organ development. Methods Fifteen single-nucleotide polymorphisms (SNPs) across TRAF6 were evaluated in 7,490 SLE patients and 6,780 control subjects from different ancestries. Population-based casecontrol association analyses and meta-analyses were performed. P values, false discovery rate q values, and odds ratios (ORs) with 95% confidence intervals (95% CIs) were calculated. Results Evidence of associations was detected in multiple SNPs. The best overall P values were obtained for SNPs rs5030437 and rs4755453 (P = 7.85 x 10(-5) and P = 4.73 x 10(-5), respectively) without significant heterogeneity among populations (P = 0.67 and P = 0.50, respectively, in Q statistic). In addition, SNP rs540386, which was previously reported to be associated with rheumatoid arthritis (RA), was found to be in linkage disequilibrium with these 2 SNPs (r2 = 0.95) and demonstrated evidence of association with SLE in the same direction (meta-analysis P = 9.15 x 10(-4), OR 0.89 [95% CI 0.830.95]). The presence of thrombocytopenia improved the overall results in different populations (meta-analysis P = 1.99 x 10(-6), OR 0.57 [95% CI 0.450.72], for rs5030470). Finally, evidence of family-based association in 34 African American pedigrees with the presence of thrombocytopenia was detected in 1 available SNP (rs5030437) with a Z score magnitude of 2.28 (P = 0.02) under a dominant model. Conclusion Our data indicate the presence of association of TRAF6 with SLE, consistent with the previous report of association with RA. These data provide further support for the involvement of TRAF6 in the pathogenesis of autoimmunity.

National Category
Medical and Health Sciences
Identifiers
urn:nbn:se:uu:diva-176803 (URN)10.1002/art.34361 (DOI)000304522100029 ()
Available from: 2012-06-27 Created: 2012-06-26 Last updated: 2017-12-07Bibliographically approved
Lessard, C. J., Adrianto, I., Ice, J. A., Wiley, G. B., Kelly, J. A., Glenn, S. B., . . . Moser, K. L. (2012). Identification of IRF8, TMEM39A, and IKZF3-ZPBP2 as Susceptibility Loci for Systemic Lupus Erythematosus in a Large-Scale Multiracial Replication Study. American Journal of Human Genetics, 90(4), 648-660
Open this publication in new window or tab >>Identification of IRF8, TMEM39A, and IKZF3-ZPBP2 as Susceptibility Loci for Systemic Lupus Erythematosus in a Large-Scale Multiracial Replication Study
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2012 (English)In: American Journal of Human Genetics, ISSN 0002-9297, E-ISSN 1537-6605, Vol. 90, no 4, p. 648-660Article in journal (Refereed) Published
Abstract [en]

Systemic lupus erythematosus (SLE) is a chronic heterogeneous autoimmune disorder characterized by the loss of tolerance to self-antigens and dysregulated interferon responses. The etiology of SLE is complex, involving both heritable and environmental factors. Candidate-gene studies and genome-wide association (GWA) scans have been successful in identifying new loci that contribute to disease susceptibility; however, much of the heritable risk has yet to be identified. In this study, we sought to replicate 1,580 variants showing suggestive association with SLE in a previously published GWA scan of European Americans; we tested a multiethnic population consisting of 7,998 SLE cases and 7,492 controls of European, African American, Asian, Hispanic, Gullah, and Amerindian ancestry to find association with the disease. Several genes relevant to immunological pathways showed association with SLE. Three loci exceeded the genome-wide significance threshold: interferon regulatory factor 8 (IRF8; rs11644034; p(meta-Euro) = 2.08 x 10(-10)), transmembrane protein 39A (TMEM39A; rs1132200; p(meta-all) 8.62 x 10(-9)), and 17q21 (rs1453560; p(meta-all) = 3.48 x 10(-10)) between IKAROS family of zinc finger 3 (AIOLOS; IKZF3) and zona pellucida binding protein 2 (ZPBP2). Fine mapping, resequencing, imputation, and haplotype analysis of IRF8 indicated that three independent effects tagged by rs8046526, rs450443, and rs4843869, respectively, were required for risk in individuals of European ancestry. Eleven additional replicated effects (5 x 10(-8) < p(meta-Euro) < 9.99 x 10(-5)) were observed with CFHR1, CADM2, LOC730109/IL12A, LPP, LOC63920, SLU7, ADAMTSL1, C10orf64, OR8D4 FAM19A2, and STXBP6. The results of this study increase the number of confirmed SLE risk loci and identify others warranting further investigation.

National Category
Medical and Health Sciences
Identifiers
urn:nbn:se:uu:diva-174377 (URN)10.1016/j.ajhg.2012.02.023 (DOI)000302833400007 ()
Available from: 2012-05-22 Created: 2012-05-15 Last updated: 2017-12-07Bibliographically approved
Darmanis, S., Gallant, C. & Landegren, U. (2012). PCR-Based Multiparametric Assays in Single Cells.. Clinical Chemistry, 58(12), 1618-1619
Open this publication in new window or tab >>PCR-Based Multiparametric Assays in Single Cells.
2012 (English)In: Clinical Chemistry, ISSN 0009-9147, E-ISSN 1530-8561, Vol. 58, no 12, p. 1618-1619Article in journal, Editorial material (Refereed) Published
National Category
Medical and Health Sciences
Identifiers
urn:nbn:se:uu:diva-187872 (URN)10.1373/clinchem.2012.195867 (DOI)000311981600002 ()23071363 (PubMedID)
Available from: 2012-12-11 Created: 2012-12-11 Last updated: 2017-12-07Bibliographically approved
Namjou, B., Kothari, P. H., Kelly, J. A., Glenn, S. B., Ojwang, J. O., Adler, A., . . . Atkinson, J. P. (2011). Evaluation of the TREX1 gene in a large multi-ancestral lupus cohort. Genes and Immunity, 12(4), 270-279
Open this publication in new window or tab >>Evaluation of the TREX1 gene in a large multi-ancestral lupus cohort
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2011 (English)In: Genes and Immunity, ISSN 1466-4879, E-ISSN 1476-5470, Vol. 12, no 4, p. 270-279Article in journal (Refereed) Published
Abstract [en]

Systemic lupus erythematosus (SLE) is a prototypic autoimmune disorder with a complex pathogenesis in which genetic, hormonal and environmental factors have a role. Rare mutations in the TREX1 gene, the major mammalian 3'-5' exonuclease, have been reported in sporadic SLE cases. Some of these mutations have also been identified in a rare pediatric neurological condition featuring an inflammatory encephalopathy known as Aicardi-Goutieres syndrome (AGS). We sought to investigate the frequency of these mutations in a large multi-ancestral cohort of SLE cases and controls. A total of 40 single-nucleotide polymorphisms (SNPs), including both common and rare variants, across the TREX1 gene, were evaluated in similar to 8370 patients with SLE and similar to 7490 control subjects. Stringent quality control procedures were applied, and principal components and admixture proportions were calculated to identify outliers for removal from analysis. Population-based case-control association analyses were performed. P-values, false-discovery rate q values, and odds ratios (OR) with 95% confidence intervals (CI) were calculated. The estimated frequency of TREX1 mutations in our lupus cohort was 0.5%. Five heterozygous mutations were detected at the Y305C polymorphism in European lupus cases but none were observed in European controls. Five African cases incurred heterozygous mutations at the E266G polymorphism and, again, none were observed in the African controls. A rare homozygous R114H mutation was identified in one Asian SLE patient, whereas all genotypes at this mutation in previous reports for SLE were heterozygous. Analysis of common TREX1 SNPs (minor allele frequency (MAF)>10%) revealed a relatively common risk haplotype in European SLE patients with neurological manifestations, especially seizures, with a frequency of 58% in lupus cases compared with 45% in normal controls (P = 0.0008, OR = 1.73, 95% CI = 1.25-2.39). Finally, the presence or absence of specific autoantibodies in certain populations produced significant genetic associations. For example, a strong association with anti-nRNP was observed in the European cohort at a coding synonymous variant rs56203834 (P = 2.99E-13, OR = 5.2, 95% CI = 3.18-8.56). Our data confirm and expand previous reports and provide additional support for the involvement of TREX1 in lupus pathogenesis. Genes and Immunity (2011) 12, 270-279; doi:10.1038/gene.2010.73; published online 27 January 2011

Keywords
autoimmunity, SLE, TREX1
National Category
Medical and Health Sciences
Identifiers
urn:nbn:se:uu:diva-155252 (URN)10.1038/gene.2010.73 (DOI)000291206100004 ()
Available from: 2011-06-20 Created: 2011-06-20 Last updated: 2017-12-11Bibliographically approved
Johansson, P., Schmidt, L., Baskaran, S., Kundu, S., Gallant, C. J., Kling, T., . . . Nelander, S.Decoding glioblastoma drug responses using an open access library of patient derived cell models.
Open this publication in new window or tab >>Decoding glioblastoma drug responses using an open access library of patient derived cell models
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(English)Manuscript (preprint) (Other academic)
Keywords
glioblastoma, GBM, precision medicine, drug response predictions, proteasome inhibitors, bortezomib
National Category
Medical Biotechnology (with a focus on Cell Biology (including Stem Cell Biology), Molecular Biology, Microbiology, Biochemistry or Biopharmacy)
Research subject
Biology with specialization in Molecular Biology; Oncology; Bioinformatics; Medical Science
Identifiers
urn:nbn:se:uu:diva-340308 (URN)
Available from: 2018-02-04 Created: 2018-02-04 Last updated: 2018-02-04
Al-Amin, A., Gallant, C., Lööf, S., Lengqvist, J., Bacanu, S., Nordlund, P. & Landegren, U.Sensitive Measurement of Cellular Drug-Target Engagement Using Multiplex Proximity Extension Assays.
Open this publication in new window or tab >>Sensitive Measurement of Cellular Drug-Target Engagement Using Multiplex Proximity Extension Assays
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(English)Manuscript (preprint) (Other academic)
National Category
Medical Biotechnology (with a focus on Cell Biology (including Stem Cell Biology), Molecular Biology, Microbiology, Biochemistry or Biopharmacy)
Identifiers
urn:nbn:se:uu:diva-366640 (URN)
Available from: 2018-11-22 Created: 2018-11-22 Last updated: 2018-11-23Bibliographically approved
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