Developments in Hematopoietic Stem Cell Expansion and Gene Editing Technologies

Yücel, Doğacan; Kocabaş, Fatih

doi:10.1007/5584_2017_114

Cited by 19 publications

(20 citation statements)

References 179 publications

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“…HSCs are recognized by their self-renewal and differentiation potential to all blood cell types. They are responsible for generation of billions of mature blood lineages throughout life of an adult 5,34 . HSC transplantation has been used for decades in the treatment of leukemia, lymphoma, some solid cancers, autoimmune diseases and inherited diseases like sickle cell anemia [41][42][43][44][45] .…”

Section: Discussionmentioning

confidence: 99%

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Development of Small Molecule MEIS Inhibitors that modulate HSC activity

Turan

Albayrak

Uslu

et al. 2020

Sci Rep

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Meis1, which belongs to TALE-type class of homeobox gene family, appeared as one of the key regulators of hematopoietic stem cell (HSC) self-renewal and a potential therapeutical target. However, small molecule inhibitors of MEIS1 remained unknown. This led us to develop inhibitors of MEIS1 that could modulate HSC activity. To this end, we have established a library of relevant homeobox family inhibitors and developed a high-throughput in silico screening strategy against homeodomain of MEIS proteins using the AutoDock Vina and PaDEL-ADV platform. We have screened over a million druggable small molecules in silico and selected putative MEIS inhibitors (MEISi) with no predicted cytotoxicity or cardiotoxicity. This was followed by in vitro validation of putative MEIS inhibitors using MEIS dependent luciferase reporter assays and analysis in the ex vivo HSC assays. We have shown that small molecules named MEISi-1 and MEISi-2 significantly inhibit MEIS-luciferase reporters in vitro and induce murine (LSKCD34 l°w cells) and human (CD34 + , CD133 + , and ALDH hi cells) HSC self-renewal ex vivo. In addition, inhibition of MEIS proteins results in downregulation of Meis1 and MEIS1 target gene expression including Hif-1α, Hif-2α and HSC quiescence modulators. MEIS inhibitors are effective in vivo as evident by induced HSC content in the murine bone marrow and downregulation of expression of MEIS target genes. These studies warrant identification of first-in-class MEIS inhibitors as potential pharmaceuticals to be utilized in modulation of HSC activity and bone marrow transplantation studies. Meis1 is a member of TALE class of transcription factors 1. Through interaction domains in the N terminus, MEIS1 cooperates in transcription factors PBX1 and HOXA9 to transactivate target genes 2,3. MEIS2 and MEIS3 protein sequences demonstrate a high degree of similarity with MEIS1 4. Meis1 was first described in leukemia mouse model and identified as a viral integration site (reviewed in 5). MEIS proteins are characterized by PBX interaction domains and a highly conserved homeodomain (HD). MEIS1 HD shares identical MEIS2 HD amino acid sequence. Studies to understand how MEIS1 HD interacts with DNA led to crystallization of MEIS1 HD with target DNA and identification of DNA sequence preferentially bound by MEIS proteins as "TGACAG" 6-8. Meis1 is highly expressed in the bone marrow 2,9. Lethality occurs in Meis1 knockout mice at mid gestation (E14.5-15.5) with a number of hematopoietic, vascular and cardiac abnormalities 10-12. Conditional and tissue specific deletion of Meis1 in bone marrow led to loss of HSC quiescence associated with expansion of HSC pool in vivo 13. Meis1 has been shown to regulate HSC metabolism through transcriptional regulation of hypoxia factors including Hif1a and Hif2a 13-16. Deletion of Meis1 or Hif-1α in HSCs leads to reduction in the cytoplasmic glycolysis and induction of mitochondrial phosphorylation. Intriguingly, studies showed a fundamental role of Meis1 in neonatal cardiac regeneration. Increased Me...

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Section: Discussionmentioning

confidence: 99%

“…We have previously outlined a number of molecular pathways under regulation of MEIS homeodomain 5,[13][14][15][16][17]27,28,[33][34][35] . We have shown that MEIS1 transcriptionally activate Hif-1α and Hif-2α expression HSCs.…”

Section: Meis Inhibitors Downregulated Meis Target Gene Expressionmentioning

confidence: 99%

Development of Small Molecule MEIS Inhibitors that modulate HSC activity

Turan

Albayrak

Uslu

et al. 2020

Sci Rep

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Section: Meis Inhibitors Downregulated Meis Target Gene Expressionmentioning

confidence: 99%

Development of Small Molecule MEIS Inhibitors that modulate HSC activity

Turan

Albayrak

Uslu

et al. 2020

Preprint

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AbstractMeis1, which belongs to TALE-type class of homeobox gene family, appeared as one of the key regulators of hematopoietic stem cell (HSC) self-renewal and a potential therapeutical target. However, small molecule inhibitors of MEIS1 remained unknown. This led us to develop inhibitors of MEIS1 that could modulate HSC activity. To this end, we have established a library of relevant homeobox family inhibitors and developed a high-throughput in silico screening strategy against homeodomain of MEIS proteins using the AutoDock Vina and PaDEL-ADV platform. We have screened over a million druggable small molecules in silico and selected putative MEIS inhibitors (MEISi) with no predicted cytotoxicity or cardiotoxicity. This was followed by in vitro validation of putative MEIS inhibitors using MEIS dependent luciferase reporter assays and analysis in the ex vivo HSC assays. We have shown that small molecules named MEISi-1 and MEISi-2 significantly inhibit MEIS-luciferase reporters in vitro and induce murine (LSKCD34low cells) and human (CD34+, CD133+, and ALDHhi cells) HSC self-renewal ex vivo. In addition, inhibition of MEIS proteins results in downregulation of Meis1 and MEIS1 target gene expression including Hif-1α, Hif-2α and HSC quiescence modulators. MEIS inhibitors are effective in vivo as evident by induced HSC content in the murine bone marrow and downregulation of expression of MEIS target genes. These studies warrant identification of first-in-class MEIS inhibitors as potential pharmaceuticals to be utilized in modulation of HSC activity and bone marrow transplantation studies.

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“…This problem suggests that additional methods and growth factors for expansion of HSCs are required in vitro studies to tackle this limitation. 3,4 It is reported that using frozen HSCs separated from umbilical cord blood (UCB) after 20 years had no negative effect on cell viability and function in HSCs transplant. Although the side effects associated with dimethyl sulfoxide (DMSO) are already recognized, it is still being used as the top standard cryoprotectant in cryoprotection used in UCB banking.…”

Section: Introductionmentioning

confidence: 99%

<p>GFc7 as a Smart Growth Nanofactor for ex vivo Expansion and Cryoprotection of Humans’ Hematopoietic Stem Cells</p>

Hafizi¹,

Kalanaky²,

Fakharzadeh³

et al. 2020

IJN

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Background: Nowadays, smart synthesized nanostructures have attracted wide attention in the field of stem cell nanotechnology due to their effect on different properties of stem cells. Methods: GFc7 growth nanofactor was synthesized based on nanochelating technology as an iron-containing copper chelator nanocomplex. The effect of this nanocomplex on the expansion and differentiation of hematopoietic stem cells (HSCs) as well as its performance as a cryoprotectant was evaluated in the present study. Results: The results showed that the absolute count of CD34 + and CD34 + CD38 − cells on days 4, 7, 10 and 13; the percentage of lactate dehydrogenase enzyme on the same days and CD34 + CXCR4 population on day 10 were significantly increased when they were treated with GFc7 growth nanofactor in a fetal bovine serum (FBS)-free medium. This medium also led to delayed differentiation in HSCs. One noticeable result was that CD34 + CD38 − cells cultured in an FBS medium were immediately differentiated into CD34 + CD38 + cells, while CD34 + CD38 − cells treated with GFc7 growth nanofactor in FBS medium did not show such an immediate significant differentiation. De-freezing GFc7-treated CD34 + cells, which were already frozen according to cord blood bank protocols, showed a higher percentage of cell viability and a larger number of colonies according to colony-forming cell assay as compared to control. Conclusion: It can be claimed that treating HSCs with GFc7 growth nanofactor leads to quality and quantity improvement of HSCs, both in terms of expansion in vitro and freezing and de-freezing processes.

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Developments in Hematopoietic Stem Cell Expansion and Gene Editing Technologies

Cited by 19 publications

References 179 publications

Development of Small Molecule MEIS Inhibitors that modulate HSC activity

Development of Small Molecule MEIS Inhibitors that modulate HSC activity

Development of Small Molecule MEIS Inhibitors that modulate HSC activity

<p>GFc7 as a Smart Growth Nanofactor for ex vivo Expansion and Cryoprotection of Humans’ Hematopoietic Stem Cells</p>

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