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Targeting NETs usin...
Targeting NETs using dual-active DNase1 variants
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- Englert, Hanna (författare)
- Univ Med Ctr Hamburg Eppendorf, Inst Clin Chem & Lab Med, Hamburg, Germany.
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- Goebel, Josephine (författare)
- Univ Med Ctr Hamburg Eppendorf, Inst Clin Chem & Lab Med, Hamburg, Germany.
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- Khong, Danika (författare)
- Univ Med Ctr Hamburg Eppendorf, Inst Clin Chem & Lab Med, Hamburg, Germany.
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- Omidi, Maryam (författare)
- Univ Med Ctr Hamburg Eppendorf, Inst Clin Chem & Lab Med, Hamburg, Germany.
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- Wolska, Nina (författare)
- Univ Med Ctr Hamburg Eppendorf, Inst Clin Chem & Lab Med, Hamburg, Germany.
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- Konrath, Sandra (författare)
- Univ Med Ctr Hamburg Eppendorf, Inst Clin Chem & Lab Med, Hamburg, Germany.
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- Frye, Maike (författare)
- Univ Med Ctr Hamburg Eppendorf, Inst Clin Chem & Lab Med, Hamburg, Germany.
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- Mailer, Reiner K. (författare)
- Univ Med Ctr Hamburg Eppendorf, Inst Clin Chem & Lab Med, Hamburg, Germany.
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- Beerens, Manu (författare)
- Univ Med Ctr Hamburg Eppendorf, Inst Clin Chem & Lab Med, Hamburg, Germany.
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- Gerwers, Julian C. (författare)
- Univ Med Ctr Hamburg Eppendorf, Inst Clin Chem & Lab Med, Hamburg, Germany.
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- Preston, Roger J. S. (författare)
- Royal Coll Surgeons Ireland, Sch Pharm & Biomol Sci, Irish Ctr Vasc Biol, Dublin, Ireland.
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- Odeberg, Jacob, Professor, 1963- (författare)
- KTH,Science for Life Laboratory, SciLifeLab,Proteinvetenskap,Arctic Univ Norway, Dept Clin Med, Tromso, Norway.
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- Butler, Lynn M. (författare)
- Karolinska Institutet,KTH,Science for Life Laboratory, SciLifeLab,Proteinvetenskap,Univ Med Ctr Hamburg Eppendorf, Inst Clin Chem & Lab Med, Hamburg, Germany.;Arctic Univ Norway, Dept Clin Med, Tromso, Norway.;Karolinska Inst, Dept Mol Med & Surg, Stockholm, Sweden.
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- Maas, Coen (författare)
- Univ Utrecht, Univ Med Ctr Utrecht, Dept Clin Chem & Haematol, Utrecht, Netherlands.
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- Stavrou, Evi X. (författare)
- Louis Stokes Vet Adm Med Ctr, Med Serv, Sect Hematol Oncol, Cleveland, OH USA.;Case Western Reserve Univ, Sch Med, Dept Med, Hematol & Oncol Div, Cleveland, OH USA.
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- Fuchs, Tobias A. (författare)
- Univ Med Ctr Hamburg Eppendorf, Inst Clin Chem & Lab Med, Hamburg, Germany.;Neutrolis Inc, Cambridge, MA USA.
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- Renne, Thomas (författare)
- Univ Med Ctr Hamburg Eppendorf, Inst Clin Chem & Lab Med, Hamburg, Germany.;Royal Coll Surgeons Ireland, Sch Pharm & Biomol Sci, Irish Ctr Vasc Biol, Dublin, Ireland.;Johannes Gutenberg Univ Mainz, Med Ctr, Ctr Thrombosis & Hemostasis CTH, Mainz, Germany.
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Univ Med Ctr Hamburg Eppendorf, Inst Clin Chem & Lab Med, Hamburg, Germany Royal Coll Surgeons Ireland, Sch Pharm & Biomol Sci, Irish Ctr Vasc Biol, Dublin, Ireland. (creator_code:org_t)
- Frontiers Media SA, 2023
- 2023
- Engelska.
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Ingår i: Frontiers in Immunology. - : Frontiers Media SA. - 1664-3224. ; 14
- Relaterad länk:
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https://doi.org/10.3...
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https://urn.kb.se/re...
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https://doi.org/10.3...
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http://kipublication...
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Abstract
Ämnesord
Stäng
- BackgroundNeutrophil Extracellular Traps (NETs) are key mediators of immunothrombotic mechanisms and defective clearance of NETs from the circulation underlies an array of thrombotic, inflammatory, infectious, and autoimmune diseases. Efficient NET degradation depends on the combined activity of two distinct DNases, DNase1 and DNase1-like 3 (DNase1L3) that preferentially digest double-stranded DNA (dsDNA) and chromatin, respectively.MethodsHere, we engineered a dual-active DNase with combined DNase1 and DNase1L3 activities and characterized the enzyme for its NET degrading potential in vitro. Furthermore, we produced a mouse model with transgenic expression of the dual-active DNase and analyzed body fluids of these animals for DNase1 and DNase 1L3 activities. We systematically substituted 20 amino acid stretches in DNase1 that were not conserved among DNase1 and DNase1L3 with homologous DNase1L3 sequences.ResultsWe found that the ability of DNase1L3 to degrade chromatin is embedded into three discrete areas of the enzyme's core body, not the C-terminal domain as suggested by the state-of-the-art. Further, combined transfer of the aforementioned areas of DNase1L3 to DNase1 generated a dual-active DNase1 enzyme with additional chromatin degrading activity. The dual-active DNase1 mutant was superior to native DNase1 and DNase1L3 in degrading dsDNA and chromatin, respectively. Transgenic expression of the dual-active DNase1 mutant in hepatocytes of mice lacking endogenous DNases revealed that the engineered enzyme was stable in the circulation, released into serum and filtered to the bile but not into the urine.ConclusionTherefore, the dual-active DNase1 mutant is a promising tool for neutralization of DNA and NETs with potential therapeutic applications for interference with thromboinflammatory disease states.
Ämnesord
- NATURVETENSKAP -- Biologi -- Biokemi och molekylärbiologi (hsv//swe)
- NATURAL SCIENCES -- Biological Sciences -- Biochemistry and Molecular Biology (hsv//eng)
Nyckelord
- neutrophil extracellular traps (NETs)
- NETosis
- NET degradation
- DNase1
- DNase1-like 3
- thromboinflammation
- protein engineering
- recombinant proteins
Publikations- och innehållstyp
- ref (ämneskategori)
- art (ämneskategori)
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Englert, Hanna
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Goebel, Josephin ...
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Khong, Danika
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Omidi, Maryam
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Wolska, Nina
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Konrath, Sandra
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visa fler...
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Frye, Maike
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Mailer, Reiner K ...
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Beerens, Manu
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Gerwers, Julian ...
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Preston, Roger J ...
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Odeberg, Jacob, ...
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Butler, Lynn M.
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Maas, Coen
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Stavrou, Evi X.
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Fuchs, Tobias A.
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Renne, Thomas
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- NATURVETENSKAP
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NATURVETENSKAP
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och Biologi
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och Biokemi och mole ...
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Frontiers in Imm ...
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Kungliga Tekniska Högskolan
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Karolinska Institutet