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Differential tissue specific expression of Kif23 alternative transcripts in mice with the human mutation causing congenital dyserythropoietic anemia type III
Umeå University, Faculty of Medicine, Department of Medical Biosciences, Medical and Clinical Genetics.
Umeå University, Faculty of Medicine, Department of Medical Biosciences, Medical and Clinical Genetics.
Umeå University, Faculty of Medicine, Department of Medical Biosciences, Medical and Clinical Genetics.ORCID iD: 0000-0001-8741-0616
2020 (English)In: Blood Cells, Molecules & Diseases, ISSN 1079-9796, E-ISSN 1096-0961, Vol. 85, article id 102483Article in journal (Refereed) Published
Abstract [en]

Kinesin Family Member 23 (KIF23), a cell cycle regulator, has a key task in cytokinesis. KIF23 over-expression in cancer has been associated with tumor growth, progression, and poor prognosis, indicating a potential to be a cancer biomarker. A mutation in KIF23 (c.2747C > G, p.P916R) was shown to cause congenital dyserythropoietic anemia, type III (CDA III). To-date, fifteen KIF23 transcripts have been annotated, but their expression is poorly investigated. We hypothesized that tissue specific expression of a particular transcript can be critical for CDA III phenotype. In this study, we quantified expression of alternative Kif23 transcripts in a mouse model with human KIF23 mutation and investigated its association with a regulator of alternative splicing, serine/arginine-rich splicing factor 3 (Srsf3). We confirmed presence of an additional exon 8 in both human and mouse KIF23 transcripts. A transcript lacking exons 17 and 18 was ubiquitously expressed in mice while other isoforms were common in human tissues however in bone marrow of knock-in mice a transcript without exon 18 was prevalent as it was in bone marrow of a CDA III patient. We conclude that the possibility that the tissue specific expression of KIF23 alternative transcripts influence the CDA III phenotype cannot be neglected.

Place, publisher, year, edition, pages
Elsevier, 2020. Vol. 85, article id 102483
Keywords [en]
KIF23, Srsf3, CDA III, Expression, Alternative splicing, Droplet digital PCR (ddPCR), Knock-in (KI) mice
National Category
Hematology
Identifiers
URN: urn:nbn:se:umu:diva-175847DOI: 10.1016/j.bcmd.2020.102483ISI: 000571001800002PubMedID: 32818800Scopus ID: 2-s2.0-85089430369OAI: oai:DiVA.org:umu-175847DiVA, id: diva2:1476704
Available from: 2020-10-15 Created: 2020-10-15 Last updated: 2023-03-24Bibliographically approved
In thesis
1. Mitotic Kinesin-Like Protein 1 (MKLP1/KIF23) in hereditary congenital dyserythropoietic anemia type III and in cancer
Open this publication in new window or tab >>Mitotic Kinesin-Like Protein 1 (MKLP1/KIF23) in hereditary congenital dyserythropoietic anemia type III and in cancer
2021 (English)Doctoral thesis, comprehensive summary (Other academic)
Abstract [en]

A hereditary form of autosomal dominant congenital dyserythropoietic anaemiatype III (CDA III) has been reported in four families from Sweden, Argentina, Cuba and USA. CDA III patients might experience signs of mild anaemia and some of them need occasional blood transfusions. Other clinical features seen in CDA III patients are retinal angioid streaks, monoclonal gammopathy of undetermined significance and multiple myeloma. Their bone marrow is characterised by presence of giant erythroblasts with up to 12 nuclei. Previously, CDA III was mapped to a region on chromosome 15q21-q25.

In this study we aimed to identify the genetic cause of CDA III, investigate the reasons why erythroid lineage in the patients’ bone marrow is mostly affected, and seek the explanation of increased rate of cancer in the Swedish CDA III family.

We identified the genetic cause of CDA III using targeted next generation sequencing. A novel missense mutation c.2747C>G, p.P916R in kinesin familymember 23 gene (KIF23) segregated with the disease in both the American and Swedish family, and was absent in databases of sequence variants from healthy individuals. Knock-down and rescue experiments in HeLa Kyoto cells showed that the P916R mutation caused cytokinesis failure which resulted in large cells with several nuclei. This was consistent with the CDA III phenotype.

To reveal interaction partners of wild-type and mutant KIF23 proteins, pull-down experiments followed by mass spectrometry and Western blot analysis were performed. This identified Coatomer Protein Complex I (COPI), a vesicle forming complex responsible for intracellular transport, as a KIF23 interactor. By using immunofluorescence and fluorescence microscopy, we showed that COPI subunits COPα and COPβ localize to the midbody during cytokinesis. These findings indicate involvement of vesicle transport proteins in mitosis and cytokinesis, though the significance of COPI-KIF23 interaction in cell division remains to be uncovered.

To address the question if other cells are affected by the KIF23 P916R mutation, we created a knock-in mouse model with Kif23 c.2726C>G, p.P909R, which corresponds to the human KIF23 c.2747C>G. However, the mice did not developany phenotype indicating CDA III. This result was consistent with the studies ofother CDA subtypes where mouse models failed, suggesting that CDA occur only in humans. Our study of human and mouse KIF23/Kif23 expression revealed novel, previously not annotated transcripts, one in human and two in mice. Expression analysis of total mRNA using droplet digital PCR demonstrated an extensive variation of KIF23 and Kif23 expression levels in all tissues. The shortest Kif23 transcript lacking exon 17 and 18 was prevalent in mice, while corresponding transcript in human was the least expressed.

Considering the importance of KIF23 in cytokinesis and KIF23 association with cancer, we hypothesized that somatic KIF23 mutations might be overrepresented in cancer. For this purpose, we screened KIF23 and its promoter in non-small cell lung cancer samples that previously demonstrated KIF23 overexpression. No pathogenic driving KIF23 variants were detected by Sangersequencing; however, subsequent genome-wide genotyping (SNP-array) detected gain of chromosome 15 in most cases that could possibly explain KIF23 overexpression.

Place, publisher, year, edition, pages
Umeå: Umeå Universitet, 2021. p. 59
Series
Umeå University medical dissertations, ISSN 0346-6612 ; 2117
Keywords
Alternative splicing, Cell division, COPI, Congenital dyserythropoietic anemia, Cytokinesis, Gene expression, KIF23 protein human, Cancer, MKLP1 protein mouse, Mutation
National Category
Medical Genetics
Research subject
Medical Genetics
Identifiers
urn:nbn:se:umu:diva-179788 (URN)978-91-7855-470-6 (ISBN)978-91-7855-471-3 (ISBN)
Public defence
2021-03-05, Hörsal Betula, byggnad 6M, målpunkt L, Norrlands Universitetssjukhus, Umeå, 09:00 (English)
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Note

P.g.a. covid 19 uppmanas åhörare att delta digitalt via Zoom. Länk: https://umu.zoom.us/j/65108113265

Available from: 2021-02-12 Created: 2021-02-10 Last updated: 2021-02-10Bibliographically approved

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Vikberg, Ann-LouiseMalla, SandhyaGolovleva, Irina

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