MAP-2:CD55 chimeric construct effectively modulates complement activation
© 2023 The Authors. The FASEB Journal published by Wiley Periodicals LLC on behalf of Federation of American Societies for Experimental Biology..
The complement system is a complex, tightly regulated protein cascade involved in pathogen defense and the pathogenesis of several diseases. Thus, the development of complement modulators has risen as a potential treatment for complement-driven inflammatory pathologies. The enzymatically inactive MAP-2 has been reported to inhibit the lectin pathway by competing with its homologous serine protease MASP-2. The membrane-bound complement inhibitor CD55 acts on the C3/C5 convertase level. Here, we fused MAP-2 to the four N-terminal domains of CD55 generating a targeted chimeric inhibitor to modulate complement activation at two different levels of the complement cascade. Its biological properties were compared in vitro with the parent molecules. While MAP-2 and CD55 alone showed a minor inhibition of the three complement pathways when co-incubated with serum (IC50MAP-2+CD55 1-4 = 60.98, 36.10, and 97.01 nM on the classical, lectin, and alternative pathways, respectively), MAP-2:CD551-4 demonstrated a potent inhibitory activity (IC50MAP-2:CD55 1-4 = 2.94, 1.76, and 12.86 nM, respectively). This inhibitory activity was substantially enhanced when pre-complexes were formed with the lectin pathway recognition molecule mannose-binding lectin (IC50MAP-2:CD55 1-4 = 0.14 nM). MAP-2:CD551-4 was also effective at protecting sensitized sheep erythrocytes in a classical hemolytic assay (CH50 = 13.35 nM). Finally, the chimeric inhibitor reduced neutrophil activation in full blood after stimulation with Aspergillus fumigatus conidia, as well as phagocytosis of conidia by isolated activated neutrophils. Our results demonstrate that MAP-2:CD551-4 is a potent complement inhibitor reinforcing the idea that engineered fusion proteins are a promising design strategy for identifying and developing drug candidates to treat complement-mediated diseases.
| Medienart: |
E-Artikel |
|---|
| Erscheinungsjahr: |
2023 |
|---|---|
| Erschienen: |
2023 |
| Enthalten in: |
Zur Gesamtaufnahme - volume:37 |
|---|---|
| Enthalten in: |
FASEB journal : official publication of the Federation of American Societies for Experimental Biology - 37(2023), 11 vom: 12. Nov., Seite e23256 |
| Sprache: |
Englisch |
|---|
| Beteiligte Personen: |
González-Del-Barrio, Lydia [VerfasserIn] |
|---|
| Links: |
|---|
| Anmerkungen: |
Date Completed 27.10.2023 Date Revised 12.11.2023 published: Print Citation Status MEDLINE |
|---|
| doi: |
10.1096/fj.202300571R |
|---|
| funding: |
|
|---|---|
| Förderinstitution / Projekttitel: |
|
| PPN (Katalog-ID): |
NLM363167285 |
|---|
| LEADER | 01000naa a22002652 4500 | ||
|---|---|---|---|
| 001 | NLM363167285 | ||
| 003 | DE-627 | ||
| 005 | 20231226092756.0 | ||
| 007 | cr uuu---uuuuu | ||
| 008 | 231226s2023 xx |||||o 00| ||eng c | ||
| 024 | 7 | |a 10.1096/fj.202300571R |2 doi | |
| 028 | 5 | 2 | |a pubmed24n1210.xml |
| 035 | |a (DE-627)NLM363167285 | ||
| 035 | |a (NLM)37823685 | ||
| 040 | |a DE-627 |b ger |c DE-627 |e rakwb | ||
| 041 | |a eng | ||
| 100 | 1 | |a González-Del-Barrio, Lydia |e verfasserin |4 aut | |
| 245 | 1 | 0 | |a MAP-2:CD55 chimeric construct effectively modulates complement activation |
| 264 | 1 | |c 2023 | |
| 336 | |a Text |b txt |2 rdacontent | ||
| 337 | |a ƒaComputermedien |b c |2 rdamedia | ||
| 338 | |a ƒa Online-Ressource |b cr |2 rdacarrier | ||
| 500 | |a Date Completed 27.10.2023 | ||
| 500 | |a Date Revised 12.11.2023 | ||
| 500 | |a published: Print | ||
| 500 | |a Citation Status MEDLINE | ||
| 520 | |a © 2023 The Authors. The FASEB Journal published by Wiley Periodicals LLC on behalf of Federation of American Societies for Experimental Biology. | ||
| 520 | |a The complement system is a complex, tightly regulated protein cascade involved in pathogen defense and the pathogenesis of several diseases. Thus, the development of complement modulators has risen as a potential treatment for complement-driven inflammatory pathologies. The enzymatically inactive MAP-2 has been reported to inhibit the lectin pathway by competing with its homologous serine protease MASP-2. The membrane-bound complement inhibitor CD55 acts on the C3/C5 convertase level. Here, we fused MAP-2 to the four N-terminal domains of CD55 generating a targeted chimeric inhibitor to modulate complement activation at two different levels of the complement cascade. Its biological properties were compared in vitro with the parent molecules. While MAP-2 and CD55 alone showed a minor inhibition of the three complement pathways when co-incubated with serum (IC50MAP-2+CD55 1-4 = 60.98, 36.10, and 97.01 nM on the classical, lectin, and alternative pathways, respectively), MAP-2:CD551-4 demonstrated a potent inhibitory activity (IC50MAP-2:CD55 1-4 = 2.94, 1.76, and 12.86 nM, respectively). This inhibitory activity was substantially enhanced when pre-complexes were formed with the lectin pathway recognition molecule mannose-binding lectin (IC50MAP-2:CD55 1-4 = 0.14 nM). MAP-2:CD551-4 was also effective at protecting sensitized sheep erythrocytes in a classical hemolytic assay (CH50 = 13.35 nM). Finally, the chimeric inhibitor reduced neutrophil activation in full blood after stimulation with Aspergillus fumigatus conidia, as well as phagocytosis of conidia by isolated activated neutrophils. Our results demonstrate that MAP-2:CD551-4 is a potent complement inhibitor reinforcing the idea that engineered fusion proteins are a promising design strategy for identifying and developing drug candidates to treat complement-mediated diseases | ||
| 650 | 4 | |a Journal Article | |
| 650 | 4 | |a Research Support, Non-U.S. Gov't | |
| 650 | 4 | |a CD55 | |
| 650 | 4 | |a MAP-2 | |
| 650 | 4 | |a complement inhibition | |
| 650 | 4 | |a complement regulation | |
| 650 | 4 | |a lectin pathway | |
| 650 | 7 | |a Complement System Proteins |2 NLM | |
| 650 | 7 | |a 9007-36-7 |2 NLM | |
| 650 | 7 | |a CD55 Antigens |2 NLM | |
| 650 | 7 | |a Lectins |2 NLM | |
| 650 | 7 | |a Transcription Factors |2 NLM | |
| 650 | 7 | |a Complement Inactivating Agents |2 NLM | |
| 650 | 7 | |a Mannose-Binding Protein-Associated Serine Proteases |2 NLM | |
| 650 | 7 | |a EC 3.4.21.- |2 NLM | |
| 700 | 1 | |a Pérez-Alós, Laura |e verfasserin |4 aut | |
| 700 | 1 | |a Cyranka, Leon |e verfasserin |4 aut | |
| 700 | 1 | |a Rosbjerg, Anne |e verfasserin |4 aut | |
| 700 | 1 | |a Nagy, Simon |e verfasserin |4 aut | |
| 700 | 1 | |a Prohászka, Zoltán |e verfasserin |4 aut | |
| 700 | 1 | |a Garred, Peter |e verfasserin |4 aut | |
| 700 | 1 | |a Bayarri-Olmos, Rafael |e verfasserin |4 aut | |
| 773 | 0 | 8 | |i Enthalten in |t FASEB journal : official publication of the Federation of American Societies for Experimental Biology |d 1989 |g 37(2023), 11 vom: 12. Nov., Seite e23256 |w (DE-627)NLM01261730X |x 1530-6860 |7 nnns |
| 773 | 1 | 8 | |g volume:37 |g year:2023 |g number:11 |g day:12 |g month:11 |g pages:e23256 |
| 856 | 4 | 0 | |u http://dx.doi.org/10.1096/fj.202300571R |3 Volltext |
| 912 | |a GBV_USEFLAG_A | ||
| 912 | |a GBV_NLM | ||
| 951 | |a AR | ||
| 952 | |d 37 |j 2023 |e 11 |b 12 |c 11 |h e23256 | ||