Investigation of cartilage oligomeric matrix protein (COMP) role in metastasis and T-cells of the tumor microenvironment
(2023) MOBN03 20231Degree Projects in Molecular Biology
- Popular Abstract
- Cartilage oligomeric matrix protein (COMP) as an EMT regulator
Cartilage oligomeric matrix protein (COMP) is an extracellular matrix glycoprotein made up of 5 identical monomers. COMP has been recently implicated in several cancer types like breast, prostate, colon and periampullary adenocarcinoma. A high expression of COMP has been associated with metastasis and the mechanisms behind this association were investigated in this project.
The effect of PMEPA1 (an EMT related gene) on cancer cells, was assessed by a migration assay after silencing PMEPA1 in COMP or mock expressing breast cancer cells. In addition, a western blot analysis where cancer cells were treated with TGF-β1 was performed to assess if the interaction of COMP and... (More) - Cartilage oligomeric matrix protein (COMP) as an EMT regulator
Cartilage oligomeric matrix protein (COMP) is an extracellular matrix glycoprotein made up of 5 identical monomers. COMP has been recently implicated in several cancer types like breast, prostate, colon and periampullary adenocarcinoma. A high expression of COMP has been associated with metastasis and the mechanisms behind this association were investigated in this project.
The effect of PMEPA1 (an EMT related gene) on cancer cells, was assessed by a migration assay after silencing PMEPA1 in COMP or mock expressing breast cancer cells. In addition, a western blot analysis where cancer cells were treated with TGF-β1 was performed to assess if the interaction of COMP and PMEPA1 (previously demonstrated) affects the EMT via the TGF-β/Smad pathway. Then, EMT markers (vimentin MMP9, slug and E-cadherin) were analysed between COMP and mock expressing tumor tissues ex vivo and cancer cells in vitro. Finally, the activation status of CD8+ T-cells in the presence of COMP was assessed in a co-culture (cancer cells + CD8+ T-cells) assay.
The migration rate of cancer cells was noticeably reduced upon silencing PMEPA1 compared to control cells, there was no significant difference in the number of migrated cells between COMP and mock after silencing PMEPA1. This result indicates that PMEPA1 might be enhancing the migration of cancer cells and that COMP-PMEPA1 interaction could have a combined effect on the migration of breast cancer cells. Interestingly, the interaction between COMP was previously reported. COMP expressing cells had significantly lower levels of phosphorylated Smad 2 and 3, and this could indicate that the interaction of COMP and PMEPA1 might be inducing EMT via the TGF-β/Smad pathway. The TGF-β/Smad pathway regulates EMT among other functions. On the other hand, PMEPA1 modulates the TGF-β/Smad pathway. Mesenchymal markers (vimentin, MMP9 and slug) were upregulated in COMP more than in mock expressing tumor tissues and cancer cells. This data could indicate that COMP is mediating metastasis through the induction of an EMT phenotype since vimentin offers mechanical strength to cells and MMP9 degrades the ECM while slug upregulates mesenchymal markers and supresses epithelial markers. Lastly, there was no significant difference in the necrotic and apoptotic cells between COMP and mock cancer cells, and the effect of COMP on cytotoxic T-cells is not clear from this data.
In conclusion, EMT is essential in metastasis to allow cancer cells to gain migratory and invasive characteristics. Hence according to our data, the role of COMP in metastasis could be to enhance the EMT phenotype by upregulating mesenchymal markers. Additionally, PMEPA1 could be enhancing the migration rate of breast cancer cells and, COMP-PMEPA1 interaction might be inducing the EMT via the TGF-β/Smad pathway. Lastly, more assays are still needed to analyse the effect of COMP on cytotoxic T-cells in the TME. The obtained data can be of use in the development of novel cancer biomarkers to help lessen the burden of cancer in the world.
Master’s Degree Project in Molecular Biology 60 credits 2023
Department of Biology, Lund University
Advisor: Anna Blom
Department of Translational Medicine, Division of Medical protein Chemistry, Lund University (Less)
Please use this url to cite or link to this publication:
http://lup.lub.lu.se/student-papers/record/9143393
- author
- Nduku, Yvonne Thokozile
- supervisor
- organization
- course
- MOBN03 20231
- year
- 2023
- type
- H2 - Master's Degree (Two Years)
- subject
- language
- English
- id
- 9143393
- date added to LUP
- 2024-01-03 10:42:06
- date last changed
- 2024-01-03 10:42:06
@misc{9143393, author = {{Nduku, Yvonne Thokozile}}, language = {{eng}}, note = {{Student Paper}}, title = {{Investigation of cartilage oligomeric matrix protein (COMP) role in metastasis and T-cells of the tumor microenvironment}}, year = {{2023}}, }