Abstract:
MMP13 is a primary catabolic factor involved in cartilage degradation through its ability to cleave type II collagen. Transcriptionally, MMP13 is regulated by 2 main elements; proximal promoter and distal enhancers. The aim of the study is to identify transcriptional elements that regulate the MMP13 gene in order to control the substantial rise in MMP13 expression observed in Osteoarthritis.
Methods:
Possible enhancers have been determined by using the Encyclopaedia of DNA Elements (ENCODE), based on histone modifications (Limb H3K4ME1 and Limb H3k27AC), Evolutionarily conserved sequences, fibroblast and muscle peaks, ChIP peaks for RUNX2, and also based on published data on vitamin D elements. Each possible enhancer sequence has been ligated to the silent HSP68 proximal promoter in a vector that expresses the LacZ gene. Each of these constructs has been tested in transgenic embryos at E15.5 days.
Results:
We have identified several active enhancers in distal and intronic sequences. Expressions in skeletal elements were detected in the 5th Intron, proximal promoter, and the distal enhancers at -19.4, and -21.5kb). In addition, expression was also seen in other cell types such as developing skin, tendon, and fibroblasts in other tissues. However, the sequence overlapping the highest peak of RunX2 at -29kb did not show any significant expression.
Conclusion:
Our results showed that the MMP13 is regulated at the level of transcription in at least three different regions, some of which coincide with RunX2 peak activity. The main distal enhancer is around -19 to22kb where expression goes beyond skeletal elements and includes other cell types. Intronic 5’ sequence reporter to respond to1l-1also show exclusively skeletal expression.
Biography:
Dr. Sara Ibrahim AlSalhi is a distinguished expert in the field of bioengineering, with a significant focus on musculoskeletal and aging sciences. She currently holds a PhD in Bioengineering from the Department of Musculoskeletal and Ageing Science at the Institute of Life Course and Medical Sciences, part of the Faculty of Health and Life Sciences. Her research interests span several critical areas, including bioengineering, bioinformatics, biochemistry, and molecular biology, with a specialized emphasis on the remodeling of bone and cartilage in musculoskeletal systems.
Dr. AlSalhi's academic journey is marked by her deep commitment to advancing biomedical sciences. She earned her Master of Science (Engineering) in Biomedical Engineering from the School of Engineering at the University of Liverpool, United Kingdom. During her MSc, she conducted pioneering research on developing nanoparticle sensor technology for real-time in vitro applications, specifically studying human mesenchymal stem cells for bone and cartilage growth within a 3D hydrogel system. This work underscored her expertise in biomedical engineering and her innovative approach to integrating nanotechnology with cellular biology.
Her research portfolio is diverse and multidisciplinary, incorporating advanced techniques in bioinformatics, such as Encode UCSC, Integrative Genomics Viewer (IGV), CIIIDER, and JASPAR2022. Dr. AlSalhi is also proficient in cutting-edge methodologies, including CRISPR gene editing, transgenic mice models, cell culturing, and histology, all of which she applies to study the molecular and genetic underpinnings of musculoskeletal aging and disease, particularly osteoarthritis.
Dr. AlSalhi is an active participant in the scientific community, contributing her expertise at numerous prestigious conferences and events. She presented at the "Cutting Edge Osteoarthritis" conference held at St Hilda’s College, University of Oxford, in July 2022, and the British Society for Matrix Biology in September 2022 in Liverpool. Her work was also showcased at the Faculty of Health & Life Sciences PGR Poster Day at Crypt Hall, Liverpool Metropolitan Cathedral, in November 2022.
Further demonstrating her leadership in the field, Dr. AlSalhi participated in the 2nd and 3rd Editions of the International Conference on Tissue Engineering and Regenerative Medicine, held in September 2022 and August 2023, respectively. Additionally, she was involved in the European Calcified Tissue Society (ECTS) 2023 Congress and the Bone Research Society (BRS) Committee meeting in Liverpool in April 2023.
Dr. AlSalhi's work continues to push the boundaries of our understanding of musculoskeletal systems, with a focus on uncovering new therapeutic strategies for age-related diseases. Her research not only advances the field but also holds significant potential for improving the quality of life for individuals suffering from musculoskeletal conditions.
