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Updated: May 1, 2026
Generation and Expansion of Primary, Malignant Pleural Mesothelioma Tumor Lines
Published on: April 21, 2022
Expression profiling stratifies mesothelioma tumors and signifies deregulation of spindle checkpoint pathway and
M B Suraokar1, M I Nunez2, L Diao3
1Department of Translational Molecular Pathology.
Background:
Malignant pleural mesothelioma (MPM) is a lethal neoplasm exhibiting resistance to most treatment regimens and requires effective therapeutic options. Though an effective strategy in many cancer, targeted therapy is relatively unexplored in MPM because the therapeutically important oncogenic pathways and networks in MPM are largely unknown.
Materials And Methods:
We carried out gene expression microarray profiling of 53 surgically resected MPMs tumors along with paired normal tissue. We also carried out whole transcriptomic sequence (RNA-seq) analysis on eight tumor specimens. Taqman-based quantitative Reverse-transcription polymerase chain reaction (qRT-PCR), western analysis and immunohistochemistry (IHC) analysis of mitotic arrest deficient-like 1 (MAD2L1) was carried out on tissue specimens. Cell viability assays of MPM cell lines were carried out to assess sensitivity to specific small molecule inhibitors.
Results:
Bioinformatics analysis of the microarray data followed by pathway analysis revealed that the mitotic spindle assembly checkpoint (MSAC) pathway was most significantly altered in MPM tumors with upregulation of 18 component genes, including MAD2L1 gene. We validated the microarray data for MAD2L1 expression using quantitative qRT-PCR and western blot analysis on tissue lysates. Additionally, we analyzed expression of the MAD2L1 protein by IHC using an independent tissue microarray set of 80 MPM tissue samples. Robust clustering of gene expression data revealed three novel subgroups of tumors, with unique expression profiles, and showed differential expression of MSAC pathway genes. Network analysis of the microarray data showed the cytoskeleton/spindle microtubules network was the second-most significantly affected network. We also demonstrate that a nontaxane small molecule inhibitor, epothilone B, targeting the microtubules have great efficacy in decreasing viability of 14 MPM cell lines.
Conclusions:
Overall, our findings show that MPM tumors have significant deregulation of the MSAC pathway and the microtubule network, it can be classified into three novel molecular subgroups of potential therapeutic importance and epothilone B is a promising therapeutic agent for MPM.
Insights
Malignant pleural mesothelioma (MPM) shows altered mitotic spindle assembly checkpoint and microtubule networks. Epothilone B effectively reduced MPM cell viability, indicating its therapeutic potential.
Area of Science:
- Oncology
- Molecular Biology
- Bioinformatics
Background:
- Malignant pleural mesothelioma (MPM) is a lethal cancer with limited treatment options.
- Targeted therapy for MPM is underdeveloped due to unknown oncogenic pathways.
Purpose of the Study:
- To identify therapeutically relevant molecular pathways and networks in MPM.
- To explore novel molecular subgroups within MPM.
- To evaluate the efficacy of epothilone B in MPM.
Main Methods:
- Gene expression microarray and RNA-seq on MPM tumors and normal tissues.
- Quantitative RT-PCR, Western blot, and IHC for MAD2L1 expression.
- Cell viability assays with small molecule inhibitors.
Main Results:
- The mitotic spindle assembly checkpoint (MSAC) pathway and microtubule network are significantly altered in MPM.
- MAD2L1 gene was upregulated in MPM tumors.
- Three novel MPM molecular subgroups with distinct expression profiles were identified.
- Epothilone B demonstrated significant efficacy in reducing MPM cell line viability.
Conclusions:
- MPM exhibits significant deregulation of the MSAC pathway and microtubule network.
- MPM can be classified into three novel molecular subgroups with therapeutic implications.
- Epothilone B is a promising therapeutic agent for MPM.
