1 Chart of haematopoiesis (15) Platelets, erythrocytes, polymorphonuclear neutrophils, monocytes, macrophages, eosinophils, basophils, lymphocytes and plasma cells are the mature cells that are found in the peripheral blood. These all develop from the pluripotential stem cell that becomes committed to the myeloid or lymphoid lineage during the early stages of haematopoiesis. It is between these two phases of the cell that a HM can arise when the process of haematopoiesis becomes deregulated. Figure from Carr & Rodak, 2012 (15).

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... HMs however have a slow onset of symptoms and involve the proliferation of mature blood cells in the peripheral blood and bone marrow ( Figure 1.1) ...

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... LK0051 family is a family affected by HMs in five generations with a total of 21 known HM cases. This family was identified from the original 1970s study and there are seven different HM case subtypes (8,9). Three affected individuals were whole genome sequenced and two unaffected family members were whole exome sequenced (Table 1.1). This family is interesting due to the clear inheritance patterns between the uncle-nephew pair (LK0051-001 and LK0051-128) and their unaffected female relative (LK0051-159). The LK0051 family pedigree, in Figure 1.3, depicts this mode of inheritance. Bioinformatics analysis using an established next generation sequencing pipeline designed for familial analysis revealed 11,432 novel, rare variants in the LK0051 family (10). Of these variants, 1,755 were found to be in the uncle-nephew pair (LK0051-001 and LK0051-128) (shown in red in Figure 1.3). Gene-based bioinformatics tools enabled Nicholas Blackburn to prioritise 8 gene variants that were predicted to be damaging and likely to have an impact on protein function. These 8 gene variants were found in TNFSF9, MMP8, TDP2, LRP5, PEX6, SAC3D1, MYO1D and DDX6. The first five were considered to be more important based on the hypothesised effect of the mutations on gene function and the potential relevance of the genes to HM development (10). whole genome sequenced and two unaffected family members were whole exome sequenced. Five prioritised genes were found in the uncle nephew pair, which has been enlarged above (in red). There is a clear inheritance pattern between the uncle-nephew pair. Unaffected individuals have developed a HM phenotype whereas unaffected individuals have never suffered from any subtype of HM (10). ...

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... LK0051 family is a family affected by HMs in five generations with a total of 21 known HM cases. This family was identified from the original 1970s study and there are seven different HM case subtypes (8,9). Three affected individuals were whole genome sequenced and two unaffected family members were whole exome sequenced (Table 1.1). This family is interesting due to the clear inheritance patterns between the uncle-nephew pair (LK0051-001 and LK0051-128) and their unaffected female relative (LK0051-159). The LK0051 family pedigree, in Figure 1.3, depicts this mode of inheritance. Bioinformatics analysis using an established next generation sequencing pipeline designed for familial analysis revealed 11,432 novel, rare variants in the LK0051 family (10). Of these variants, 1,755 were found to be in the uncle-nephew pair (LK0051-001 and LK0051-128) (shown in red in Figure 1.3). Gene-based bioinformatics tools enabled Nicholas Blackburn to prioritise 8 gene variants that were predicted to be damaging and likely to have an impact on protein function. These 8 gene variants were found in TNFSF9, MMP8, TDP2, LRP5, PEX6, SAC3D1, MYO1D and DDX6. The first five were considered to be more important based on the hypothesised effect of the mutations on gene function and the potential relevance of the genes to HM development (10). whole genome sequenced and two unaffected family members were whole exome sequenced. Five prioritised genes were found in the uncle nephew pair, which has been enlarged above (in red). There is a clear inheritance pattern between the uncle-nephew pair. Unaffected individuals have developed a HM phenotype whereas unaffected individuals have never suffered from any subtype of HM (10). ...

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... variant in TNFSF9 (also referred to as CD137L throughout this study) was found to be present in the uncle and nephew pair of the LK0051 family (Figure 1.3). The TNFSF9 mutation was predicted to be damaging and deleterious by bioinformatics tools SIFT and PROVEAN, which predict whether an amino acid substitution affects protein function ...

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... mature blood cell is produced in different numbers by the body (18). The rate of blood cell production is dependent on the destruction of blood cells in response to changing body conditions and a number of regulatory factors (19). Figure 1.1 shows that all red and white blood cells originate from the pluripotent haematopoietic stem cell, which is also known as the haemocytoblast. This stem cell can become committed to either the myeloid or lymphoid lineage depending on cytokine involvement (15). Disturbances of this haematopoietic tree can lead to malignant transformations, such as HMs ...

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... lymphoma (BL) all comprise NHL (16,17). Acute is the term used to describe the fast onset of disease and is usually associated with the proliferation of blast cells (Figure ...

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... occur when the body's bone marrow becomes defective and the blood cells multiply out of control (15). The normal haematopoietic process thus becomes deregulated and there is clonal expansion of a single cell (21). This causes inappropriate proliferation of the cell and at times the cells may be released from the bone marrow immaturely. These are termed blast cells, and are often seen in acute leukaemias (19). For example myeloblasts are observed in AML and a number of other acute leukaemias involving the myeloid lineage. Inappropriate proliferation can occur in any of the stem, progenitor, precursor or peripheral cells, which are illustrated in Figure 1. 1 (15). Mutations in stem, progenitor and early precursor cells are termed acute leukaemias and mutations in the late precursor and peripheral cells are more chronic and dormant. It is by a somatic mutation in one allele of an otherwise normal proto- oncogene that causes it to become defective and differ from its precursor. This mutation is a genetic alteration that is passed to its progeny during inappropriate proliferation, which leads to the development of cancer (19). ...

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... Guinea, NHL had a prevalence of 3.76 per 100,000 people whereas in Australia it had the highest five-year prevalence per 100,000 people, with 67.48. This is probably due to the limited health resources in Guinea; this is not to say that Guinea doesn't have a prevalence of NHL as high as Australia, however it has not be been recorded. It is interesting to note that Australia and the USA have one of the highest five year prevalence for multiple myeloma (MM), leukaemia, NHL and HL (Figure 1.2) (22). However, not one HM falls in to the top five most diagnosed cancers in the USA, which could be explained by an overall higher prevalence of cancer in general. Thus it is very noticeable that the developed countries of the world, including Australia, have a higher prevalence of HMs compared to the developing world, which is supported by GLOBOCAN's estimates of HM incidence and mortality worldwide (Figure ...

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... Guinea, NHL had a prevalence of 3.76 per 100,000 people whereas in Australia it had the highest five-year prevalence per 100,000 people, with 67.48. This is probably due to the limited health resources in Guinea; this is not to say that Guinea doesn't have a prevalence of NHL as high as Australia, however it has not be been recorded. It is interesting to note that Australia and the USA have one of the highest five year prevalence for multiple myeloma (MM), leukaemia, NHL and HL (Figure 1.2) (22). However, not one HM falls in to the top five most diagnosed cancers in the USA, which could be explained by an overall higher prevalence of cancer in general. Thus it is very noticeable that the developed countries of the world, including Australia, have a higher prevalence of HMs compared to the developing world, which is supported by GLOBOCAN's estimates of HM incidence and mortality worldwide (Figure ...

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... The variant (rs138686754) was prioritised in this family due to the uncle-nephew pair carrying the mutation (LK0051-001 and LK0051-128). The mode of inheritance via the carrier sister/mother is explained by the family pedigree in Figure 1.3 previously. The MMP8 mutation was predicted to be neutral and tolerated by bioinformatics tools, SIFT and PROVEAN and due to its role in bone marrow formation may be predisposing these individuals to HM development ...

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... of the Anti-TNFSF9 antibodies (Abcam, Atlas and Santa Cruz) determined there to be similar CD137L expression in one of the cell lines at baseline (Figure 4.11). Therefore any of the antibodies could be used for further analysis, as there were no discrepancies between the three when measuring CD137L ...

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... measurement of CD137L determined there to be no difference between the TNFSF9 mutated and non-mutated cell lines (Figure 4.12). This shows that the TNFSF9 mutation does not effect TNFSF9 (CD137L) expression by flow cytometry analysis. B lymphocyte cells were incubated with Fc or CD137: Fc as described in Section 4.2.10 and there was found to be no effect (Figure 4.13). Mutation and non-mutation carriers all had similar CD137L expression levels when incubated with Fc and CD137: ...