(provided by the applicant): Coagulation factors V (FV) and factor VIII (FVIII) are both secreted glycoproteins that share pivotal roles in both hemostasis and thrombosis. Although many secreted proteins (referred to as cargo) are believed to require transport receptors for efficient ER-to-Golgi transport, only a limited number of such receptors have been described, mostly in yeast. Evidence for the existence of mammalian cargo receptors came unexpectedly from studies of the human genetic disorder combined deficiency of FV and FVIII, which identified mutations in LMAN1 and MCFD2 as the cause of the disorder. Combined deficiency of FV and FVIII is a rare bleeding disorder characterized by coordinate reduction of both FV and FVIII to the range of 5-30% of normal. LMAN1 and MCFD2 form a Ca2+-dependent protein complex in the ER-Golgi intermediate compartment that interacts with FV and FVIII, suggesting that the LMAN1-MCFD2 complex is a cargo receptor required for efficient transport of FV and FVIII from the ER to the Golgi. Mice deficient in LMAN1 model the human disorder but also exhibit a strain-specific perinatal lethality that is not explained by the reduced FV/FVIII levels, indicating additional functions of LMAN1. Using LMAN1 and MCFD2 deficient mice, proposed research will elucidate structure-function relationship of the LMAN1-MCFD2 receptor complex, and its requirement for the secretion of FV and FVIII and other potential cargo proteins in vivo. Proposed studies will not only answer fundamental questions regarding the mechanism of LMAN1-MCFD2 receptor-mediated secretion of FV and FVIII, but will also provide fundamental new insights into general mechanism of mammalian ER-to-Golgi protein transport. Genetic deficiency of FVIII results in hemophilia A, which affects ~1 in 5000 males. On the other hand, a gain-of-function mutation in FV (FV Leiden) and increased FVIII activity are major risk factors for venous thrombosis, which affects ~1:1,000 individuals in the US per year. The findings will have practical importance for improving FVIII expression and may expedite the eventual goal of somatic cell gene therapy for hemophilia A, as well as new approaches to limiting FV and FVIII production in prothrombotic states.

Public Health Relevance

The project will study a transport receptor that when absent, cause a human genetic disorder with low levels of blood coagulation factors V and VIII. The goal is to understand how this transport receptor controls two different blood coagulation factors and hopefully use the knowledge to benefit people with hemophilia A and venous thrombosis.

Agency
National Institute of Health (NIH)
Institute
National Heart, Lung, and Blood Institute (NHLBI)
Type
Research Project (R01)
Project #
5R01HL094505-04
Application #
8197406
Study Section
Hemostasis and Thrombosis Study Section (HT)
Program Officer
Link, Rebecca P
Project Start
2008-12-26
Project End
2013-11-30
Budget Start
2011-12-01
Budget End
2012-11-30
Support Year
4
Fiscal Year
2012
Total Cost
$353,250
Indirect Cost
$128,250
Name
Cleveland Clinic Lerner
Department
Other Basic Sciences
Type
Schools of Medicine
DUNS #
135781701
City
Cleveland
State
OH
Country
United States
Zip Code
44195
Khoriaty, Rami; Vasievich, Matthew P; Jones, Morgan et al. (2014) Absence of a red blood cell phenotype in mice with hematopoietic deficiency of SEC23B. Mol Cell Biol 34:3721-34
Baines, Andrea C; Adams, Elizabeth J; Zhang, Bin et al. (2013) Disruption of the Sec24d gene results in early embryonic lethality in the mouse. PLoS One 8:e61114
Zheng, Chunlei; Zhang, Bin (2013) Combined deficiency of coagulation factors V and VIII: an update. Semin Thromb Hemost 39:613-20
Patel, A J; Liu, H-H; Lager, R A et al. (2013) Successful percutaneous coronary intervention in a patient with combined deficiency of FV and FVIII due to novel compound heterozygous mutations in LMAN1. Haemophilia 19:607-10
Zheng, Chunlei; Page, Richard C; Das, Vaijayanti et al. (2013) Structural characterization of carbohydrate binding by LMAN1 protein provides new insight into the endoplasmic reticulum export of factors V (FV) and VIII (FVIII). J Biol Chem 288:20499-509
Zhang, Bin; Zheng, Chunlei; Zhu, Min et al. (2011) Mice deficient in LMAN1 exhibit FV and FVIII deficiencies and liver accumulation of ýý1-antitrypsin. Blood 118:3384-91
Zheng, Chunlei; Liu, Hui-hui; Zhou, Jiahai et al. (2010) EF-hand domains of MCFD2 mediate interactions with both LMAN1 and coagulation factor V or VIII. Blood 115:1081-7
Zhang, Bin (2009) Recent developments in the understanding of the combined deficiency of FV and FVIII. Br J Haematol 145:15-23