Genetic studies into rare diseases and cancer using next generation sequencing technologies

Rare genetic diseases (RGDs) and rare sporadic cancers are often considered as two separate groups of diseases. Nevertheless, both groups share the same burden: their rarity and the challenges in diagnosis and treatment, thus affecting the wellbeing of many patients and their families around the world. Although next generation sequencing (NGS) technologies have revolutionised the genetic landscape of RGDs and cancers, many patients with these diseases are still without a definitive molecular diagnosis. In this thesis, NGS was conducted on congenital hypothyroidism (CHT) families (an example of an RGD) and three rare bone cancers, aiming to expand the understanding of the genetic and pathogenic mechanisms of these diseases. To identify known or novel disease-causing genes, WES was conducted on four families with CHT. In one family, a homozygous candidate variant in SIX2 was identified, and subsequent functional characterisation experiments and family segregation analyses were performed. After more family members were included, the SIX2 variant did not segregate with the disease in the family and, therefore, was classified as unlikely disease causing. WES and RNA sequencing (RNA-Seq) were conducted on three rare bone tumours: undifferentiated pleomorphic sarcoma of bone (UPSb), adamantinoma and osteofibrous dysplasia (OFD)-like adamantinoma. In UPSb tumours, 31 genes were recurrently mutated, including TP53 in 4/14 samples (29%), and chromatin remodelling genes (ATRX, H3F3A, DOT1L) in 5/14 samples (36%). In addition, two previously reported gene fusions (CLTC-VMP1 and FARP1-STK24) were identified in these tumours. In adamantinoma tumours, KMT2D, a histone methyltransferase, was recurrently mutated in 2/8 adamantinomas (25%). In addition, a cancer predisposing germline fusion (KANSL1-ARL17A) was identified in 4/6 adamantinoma (66.7%) and in 3/4 OFD-like adamantinoma (75%) tumours. This thesis is a practical example demonstrating how rare diseases and cancers can be investigated using the same high-throughput techniques. Moreover, the three bone tumour studies represent the first comprehensive WES and RNA-Seq analyses conducted on these tumours, revealing novel molecular insights that can be translated into clinical practices to enhance the diagnosis, prognosis and the outcomes of patients with these diseases.