The Role of Thrombopoietin Signalling in JAK2V617F-positive Myeloproliferative Neoplasms

Thrombopoietin (TPO) is the primary regulator of megakaryocyte development, regulating proliferation and differentiation in addition to the number of circulating platelets through binding to and stimulation of the cell surface receptor MPL. Activating mutations in MPL constitutively stimulate downstream signalling pathways, leading to aberrant haematopoiesis and contribute to development of myeloproliferative neoplasms (MPNs). Several studies have mapped the tyrosine residues within the cytoplasmic domain of MPL that mediate these cellular signals; however, secondary signalling pathways are incompletely understood. Additionally, the identification of the JAK2V617F mutation has profoundly increased our understanding of MPNs and although a role has been implicated in vitro, the in vivo role of MPL in JAK2V617F-positive MPNs has yet to be determined. In this thesis, a novel signalling pathway for the negative regulation of TPO signalling was identified whereby MPLY591 is phosphorylated resulting in association of SYK which negatively regulates TPO-mediated ERK1/2 signalling. Additionally, genetic manipulation of an in vivo JAK2V617F-positive MPN mouse model led to the identification of MPL as an essential molecular component for development of JAK2V617F-postive MPNs. In the absence or reduction of MPL, the disease fails to develop. However, removal of the cytokine, TPO, was unable to prevent the disease from developing. These findings provide novel insights not only into regulation of TPO-signalling but also the role of TPO and MPL in JAK2V617F-positive MPN disease pathogenesis. Identification of the role of MPL in MPN pathogenesis, as well as insights into additional regulatory pathways, contributes to our understanding of normal and pathological TPO signalling. These new insights also provide a basis for development of novel therapeutics for the treatment of MPNs and other diseases resulting from aberrant of TPO signalling

Being Thai: an exploration of the hybridity of Thai identity through contemporary art practice

This practice-led PhD research focuses on what I propose to be the inseparable relationship between my visual art practice and my understanding of social identity within the Thai context. In the process, this research contributes to a body of imagery unique to Thai cultural experience and adds to a representation of contributions on its understanding, within a national and global context. Specifically, for this research I examine the effect of geographical and political environments on what I identify as the plurality and hybridity of Thai identities. This research is undertaken through visual investigations into the historical framework within Thailand itself and individual experiences of identity experienced within Thailand and viewed from a distance

The thrombopoietin receptor : revisiting the master regulator of platelet production

Thrombopoietin (TPO) and its receptor, MPL, are the primary regulators of platelet production and critical for hematopoietic stem cell (HSC) maintenance. Since TPO was first cloned in 1994, the physiological and pathological roles of TPO and MPL have been well characterized, culminating in the first MPL agonists being approved for the treatment of chronic immune thrombocytopenia in 2008. Dysregulation of the TPO-MPL signaling axis contributes to the pathogenesis of hematological disorders: decreased expression or function results in severe thrombocytopenia progressing to bone marrow failure, while hyperactivation of MPL signaling, either by mutations in the receptor or associated Janus kinase 2 (JAK2), results in pathological myeloproliferation. Despite its importance, it was only recently that the long-running debate over the mechanism by which TPO binding activates MPL has been resolved. This review will cover key aspects of TPO and MPL structure and function and their importance in receptor activation, discuss how these are altered in hematological disorders and consider how a greater understanding could lead to the development of better-targeted and more efficacious therapies

Hepcidin, Serum Iron, and Transferrin Saturation in Full-Term and Premature Infants during the First Month of Life: A State-of-the-Art Review of Existing Evidence in Humans.

Neonates regulate iron at birth and in early postnatal life. We reviewed literature from PubMed and Ovid Medline containing data on umbilical cord and venous blood concentrations of hepcidin and iron, and transferrin saturation (TSAT), in human neonates from 0 to 1 mo of age. Data from 59 studies were used to create reference ranges for hepcidin, iron, and TSAT for full-term-birth (FTB) neonates over the first month of life. In FTB neonates, venous hepcidin increases 100% over the first month of life (to reach 61.1 ng/mL; 95% CI: 20.1, 102.0 ng/mL) compared with umbilical cord blood (29.7 ng/mL; 95% CI: 21.1, 38.3 ng/mL). Cord blood has a high concentration of serum iron (28.4 μmol/L; 95% CI: 26.0, 31.1 μmol/L) and levels of TSAT (51.7%; 95% CI: 46.5%, 56.9%). After a short-lived immediate postnatal hypoferremia, iron and TSAT rebounded to approximately half the levels in the cord by the end of the first month. There were insufficient data to formulate reference ranges for preterm neonates

Regulation of the Iron Homeostatic Hormone Hepcidin

Iron is required for many biological processes but is also toxic in excess; thus, body iron balance is maintained through sophisticated regulatory mechanisms. The lack of a regulated iron excretory mechanism means that body iron balance is controlled at the level of absorption from the diet. Iron absorption is regulated by the hepatic peptide hormone hepcidin. Hepcidin also controls iron release from cells that recycle or store iron, thus regulating plasma iron concentrations. Hepcidin exerts its effects through its receptor, the cellular iron exporter ferroportin. Important regulators of hepcidin, and therefore of systemic iron homeostasis, include plasma iron concentrations, body iron stores, infection and inflammation, and erythropoiesis. Disturbances in the regulation of hepcidin contribute to the pathogenesis of many iron disorders: hepcidin deficiency causes iron overload in hereditary hemochromatosis and nontransfused β-thalassemia, whereas overproduction of hepcidin is associated with iron-restricted anemias seen in patients with chronic kidney disease, chronic inflammatory diseases, some cancers, and inherited iron-refractory iron deficiency anemia. This review summarizes our current understanding of the molecular mechanisms and signaling pathways involved in the control of hepcidin synthesis in the liver, a principal determinant of plasma hepcidin concentrations

Placental iron transport: The mechanism and regulatory circuits

As the interface between the fetal and maternal circulation, the placenta facilitates both nutrient and waste exchange for the developing fetus. Iron is essential for healthy pregnancy, and transport of iron across the placenta is required for fetal growth and development. Perturbation of this transfer can lead to adverse pregnancy outcomes. Despite its importance, our understanding of how a large amount of iron is transported across placental membranes, how this process is regulated, and which iron transporter proteins function in different placental cells remains rudimentary. Mechanistic studies in mouse models, including placenta-specific deletion or overexpression of iron-related proteins will be essential to make progress. This review summarizes our current understanding about iron transport across the syncytiotrophoblast under physiological conditions and identifies areas for further investigation

Erythroferrone contributes to iron mobilization for embryo erythropoiesis in iron-deficient mouse pregnancies.

Erythroferrone (ERFE) is an erythroblast-secreted regulator of iron metabolism. The production of ERFE increases during stress erythropoiesis, leading to decreased hepcidin expression and mobilization of iron. Pregnancy requires a substantial increase in iron availability to sustain maternal erythropoietic expansion and fetal development and is commonly affected by iron deficiency. To define the role of ERFE during iron-replete or iron-deficient pregnancy, we utilized mouse models expressing a range of ERFE levels: transgenic (TG) mice overexpressing ERFE, wild-type (WT), and ERFE knockout (KO) mice. We altered maternal iron status using diets with low or standard iron content and performed the analysis at E18.5. Iron deficiency increased maternal ERFE in WT pregnancy. Comparing different maternal genotypes, ERFE TG dams had lower hepcidin relative to their liver iron load but similar hematological parameters to WT dams on either diet. In ERFE KO dams, most hematologic and iron parameters were comparable to WT, but mean corpuscular volume (MCV) was decreased under both iron conditions. Similar to dams, TG embryos had lower hepcidin on both diets, but their hematologic parameters did not differ from those of WT embryos. ERFE KO embryos had lower MCV than WT embryos on both diets. The effect was exacerbated under iron-deficient conditions where ERFE KO embryos had higher hepcidin, lower Hb and Hct, and lower brain iron concentration compared to WT embryos, indicative of iron restriction. Thus, under iron-deficient conditions, maternal and embryo ERFE facilitate iron mobilization for embryonic erythropoiesis