KLF4 overexpression protects against complement-mediated endothelial injury in transplant-associated thrombotic microangiopathy

Transplant-associated thrombotic microangiopathy (TA-TMA) is a severe complication of hematopoietic stem cell transplantation, marked by excessive complement activation, endothelial injury, and microangiopathy. Although complement blockade benefits some patients, effective prophylactic and therapeu...

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Main Authors: Shuhui Jiang, Jiaqian Qi, Tingting Pan, Zhenzhen Yao, Siyi Lu, Yifei Han, Depei Wu, Yue Han
Format: Article
Language:English
Published: Ferrata Storti Foundation 2025-08-01
Series:Haematologica
Online Access:https://haematologica.org/article/view/12207
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author Shuhui Jiang
Jiaqian Qi
Tingting Pan
Zhenzhen Yao
Siyi Lu
Yifei Han
Depei Wu
Yue Han
author_facet Shuhui Jiang
Jiaqian Qi
Tingting Pan
Zhenzhen Yao
Siyi Lu
Yifei Han
Depei Wu
Yue Han
author_sort Shuhui Jiang
collection DOAJ
description Transplant-associated thrombotic microangiopathy (TA-TMA) is a severe complication of hematopoietic stem cell transplantation, marked by excessive complement activation, endothelial injury, and microangiopathy. Although complement blockade benefits some patients, effective prophylactic and therapeutic strategies remain scarce. Therefore, plasma samples from 20 TA-TMA patients and 1:1 matched control patients (matched by age, sex, underlying diagnosis, HLA compatibility, graft source, and donor–recipient ABO blood type) were measured for krüppel-like factor 4 (KLF4), complement proteins and endothelial injury markers. The mechanism was investigated both in vitro and in vivo. In this study, plasma analysis revealed that TA-TMA patients exhibit notably lower KLF4 levels compared to matched controls, as well as elevated endothelial injury markers. In vitro, increased KLF4 expression in human umbilical vein endothelial cells significantly reduced complement deposition and mitigated endothelial damage induced by TA-TMA plasma. Furthermore, KLF4 overexpression notably decreased apoptosis and preserved endothelial barrier integrity. In a mouse model of TA-TMA triggered by dimethyloxalylglycine, upregulation of KLF4 alleviated anemia, thrombocytopenia, and renal complement deposition, while diminishing endothelial inflammatory and thrombotic markers. Intriguingly, pravastatin treatment produced similar improvements. Mechanistic analyses using CUT&Tag, RNA sequencing, luciferase assays, and quantitative real-time PCR revealed that KLF4 binds to the CD46 promoter, enhancing its transcription and thus restraining complement activation in endothelial cells. These results identify KLF4 as a key negative regulator of complement-mediated endothelial injury in TA-TMA. This conclusion is supported by CD46 knockdown abolishing KLF4-mediated benefits, highlighting the therapeutic potential of targeting KLF4 or its downstream effectors, including CD46.
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spelling doaj-art-d9dad013fb914c44a9c3e4bbd1a48ec32025-08-20T02:58:23ZengFerrata Storti FoundationHaematologica0390-60781592-87212025-08-01999110.3324/haematol.2025.287676KLF4 overexpression protects against complement-mediated endothelial injury in transplant-associated thrombotic microangiopathyShuhui Jiang0Jiaqian Qi1Tingting Pan2Zhenzhen Yao3Siyi Lu4Yifei Han5Depei Wu6Yue Han7National Clinical Research Center for Hematologic Diseases, Jiangsu Institute of Hematology, The First Affiliated Hospital of Soochow University, Suzhou, China; Institute of Blood and Marrow Transplantation, Collaborative Innovation Center of Hematology, Soochow University, Suzhou, China; Key Laboratory of Thrombosis and Hemostasis of Ministry of Heath, SuzhouNational Clinical Research Center for Hematologic Diseases, Jiangsu Institute of Hematology, The First Affiliated Hospital of Soochow University, Suzhou, China; Institute of Blood and Marrow Transplantation, Collaborative Innovation Center of Hematology, Soochow University, Suzhou, China; Key Laboratory of Thrombosis and Hemostasis of Ministry of Heath, SuzhouNational Clinical Research Center for Hematologic Diseases, Jiangsu Institute of Hematology, The First Affiliated Hospital of Soochow University, Suzhou, China; Institute of Blood and Marrow Transplantation, Collaborative Innovation Center of Hematology, Soochow University, Suzhou, China; Key Laboratory of Thrombosis and Hemostasis of Ministry of Heath, SuzhouNational Clinical Research Center for Hematologic Diseases, Jiangsu Institute of Hematology, The First Affiliated Hospital of Soochow University, Suzhou, China; Institute of Blood and Marrow Transplantation, Collaborative Innovation Center of Hematology, Soochow University, Suzhou, China; Key Laboratory of Thrombosis and Hemostasis of Ministry of Heath, SuzhouNational Clinical Research Center for Hematologic Diseases, Jiangsu Institute of Hematology, The First Affiliated Hospital of Soochow University, Suzhou, China; Institute of Blood and Marrow Transplantation, Collaborative Innovation Center of Hematology, Soochow University, Suzhou, China; Key Laboratory of Thrombosis and Hemostasis of Ministry of Heath, SuzhouNational Clinical Research Center for Hematologic Diseases, Jiangsu Institute of Hematology, The First Affiliated Hospital of Soochow University, Suzhou, China; Institute of Blood and Marrow Transplantation, Collaborative Innovation Center of Hematology, Soochow University, Suzhou, China; Key Laboratory of Thrombosis and Hemostasis of Ministry of Heath, SuzhouNational Clinical Research Center for Hematologic Diseases, Jiangsu Institute of Hematology, The First Affiliated Hospital of Soochow University, Suzhou, China; Institute of Blood and Marrow Transplantation, Collaborative Innovation Center of Hematology, Soochow University, Suzhou, China; Key Laboratory of Thrombosis and Hemostasis of Ministry of Heath, Suzhou, China; State Key Laboratory of Radiation Medicine and Protection, Soochow University, SuzhouNational Clinical Research Center for Hematologic Diseases, Jiangsu Institute of Hematology, The First Affiliated Hospital of Soochow University, Suzhou, China; Institute of Blood and Marrow Transplantation, Collaborative Innovation Center of Hematology, Soochow University, Suzhou, China; Key Laboratory of Thrombosis and Hemostasis of Ministry of Heath, Suzhou, China; State Key Laboratory of Radiation Medicine and Protection, Soochow University, Suzhou Transplant-associated thrombotic microangiopathy (TA-TMA) is a severe complication of hematopoietic stem cell transplantation, marked by excessive complement activation, endothelial injury, and microangiopathy. Although complement blockade benefits some patients, effective prophylactic and therapeutic strategies remain scarce. Therefore, plasma samples from 20 TA-TMA patients and 1:1 matched control patients (matched by age, sex, underlying diagnosis, HLA compatibility, graft source, and donor–recipient ABO blood type) were measured for krüppel-like factor 4 (KLF4), complement proteins and endothelial injury markers. The mechanism was investigated both in vitro and in vivo. In this study, plasma analysis revealed that TA-TMA patients exhibit notably lower KLF4 levels compared to matched controls, as well as elevated endothelial injury markers. In vitro, increased KLF4 expression in human umbilical vein endothelial cells significantly reduced complement deposition and mitigated endothelial damage induced by TA-TMA plasma. Furthermore, KLF4 overexpression notably decreased apoptosis and preserved endothelial barrier integrity. In a mouse model of TA-TMA triggered by dimethyloxalylglycine, upregulation of KLF4 alleviated anemia, thrombocytopenia, and renal complement deposition, while diminishing endothelial inflammatory and thrombotic markers. Intriguingly, pravastatin treatment produced similar improvements. Mechanistic analyses using CUT&Tag, RNA sequencing, luciferase assays, and quantitative real-time PCR revealed that KLF4 binds to the CD46 promoter, enhancing its transcription and thus restraining complement activation in endothelial cells. These results identify KLF4 as a key negative regulator of complement-mediated endothelial injury in TA-TMA. This conclusion is supported by CD46 knockdown abolishing KLF4-mediated benefits, highlighting the therapeutic potential of targeting KLF4 or its downstream effectors, including CD46. https://haematologica.org/article/view/12207
spellingShingle Shuhui Jiang
Jiaqian Qi
Tingting Pan
Zhenzhen Yao
Siyi Lu
Yifei Han
Depei Wu
Yue Han
KLF4 overexpression protects against complement-mediated endothelial injury in transplant-associated thrombotic microangiopathy
Haematologica
title KLF4 overexpression protects against complement-mediated endothelial injury in transplant-associated thrombotic microangiopathy
title_full KLF4 overexpression protects against complement-mediated endothelial injury in transplant-associated thrombotic microangiopathy
title_fullStr KLF4 overexpression protects against complement-mediated endothelial injury in transplant-associated thrombotic microangiopathy
title_full_unstemmed KLF4 overexpression protects against complement-mediated endothelial injury in transplant-associated thrombotic microangiopathy
title_short KLF4 overexpression protects against complement-mediated endothelial injury in transplant-associated thrombotic microangiopathy
title_sort klf4 overexpression protects against complement mediated endothelial injury in transplant associated thrombotic microangiopathy
url https://haematologica.org/article/view/12207
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