PTM of ceramide synthase 2 enzyme leads to tumor progression

March 5, 2021

Tumor Progression in Breast Cancer

Interview with Trishna Pani, Kajal Rajput, and Animesh Kar

Trishna Pani, Kajal Rajput, and Animesh Kar are joint first authors of the research article titled “Alternative splicing of ceramide synthase 2 alters levels of specific ceramides and modulates cancer cell proliferation and migration in Luminal B breast cancer subtype”, published in Cell Death & Disease (2021).

About the Researchers:

Trishna Pani completed her post-graduation in biotechnology from Vellore Institute of Technology and is currently pursuing her Ph.D. from Amity University, Manesar under Dr. Ujjaini Dasgupta and co-supervision of Dr. Avinash Bajaj (RCB, Faridabad). Her research focuses on post-transcriptional modifications of sphingolipid genes in different breast cancer subtypes to develop better diagnostic markers and therapies.
Kajal Rajput graduated and completed her post-graduation in biotechnology from Maharishi Dayanand University (MDU), Rohtak. She is pursuing a Ph.D. from Amity University, Gurgaon under Dr. Ujjaini Dasgupta. Her goal is to identify sphingolipid biomarkers in breast cancer subtypes and develop therapeutic strategies to target them.
Animesh Kar holds an undergraduate degree in Biomedical Science from the University of Delhi and a Master’s in Biophysics from AIIMS, Delhi. He is pursuing a Ph.D. under Dr. Avinash Bajaj at RCB, Faridabad. His research aims to understand and modulate molecular signatures in cancer using engineered nanotherapeutics.

How would you explain your paper’s key results to the non-scientific community?

Cell signaling is vital for coordinating cell behavior. Receptor-based signaling pathways control essential cellular functions, and any disruption can lead to diseases such as cancer. Among various cancers, breast cancer is the most common in females and is classified into subtypes: Luminal A, Luminal B, HER2+, and Triple Negative—each with unique gene expression and clinical behavior.

Sphingolipids are bioactive lipids that regulate many cellular processes. Alterations in sphingolipid metabolism are implicated in cancer. Our previous work showed distinct sphingolipid profiles for each breast cancer subtype.

In this study, we analyzed RNA sequencing data from the TCGA BRCA cohort and found that the enzyme Ceramide Synthase 2 (CERS2) undergoes alternative splicing (exon 8 skipping) specifically in Luminal B breast cancer. CERS2 synthesizes very-long-chain ceramides, which inhibit tumor cell proliferation and metastasis. The skipped exon (exon 8) encodes the Lag1P domain, critical for substrate recognition.

Loss of exon 8 alters the enzyme’s product profile, reducing protective ceramides and promoting cancer cell proliferation and migration. We validated this in Indian Luminal B tumor samples and their derived cell lines.

What are the possible consequences of these findings for your research area?

Our research highlights the role of sphingolipid metabolism in the tumor microenvironment. Given the significant role of CERS2, we plan to study its expression in a larger patient cohort. Positive findings may help develop this splicing event into a diagnostic marker or even a therapeutic target.

[…] we have interesting results related to CERS2, it is very important to see its role in a large patient cohort.

What was the exciting moment (eureka moment) during your research?

Trishna Pani: My most exciting moment was detecting both isoforms of CERS2 via western blotting in Indian tumor samples—and seeing higher abundance of the spliced version in tumors versus controls.
Kajal Rajput: The eureka moment came when the phenotypic effects of overexpressing the spliced isoform in cancer cell lines matched our hypothesis.
Animesh Kar: For me, it was confirming the existence of the spliced transcript via qRT-PCR, with band sizes exactly as predicted.

What do you hope to do next?

We aim to investigate the mechanistic basis of this post-transcriptional event in CERS2 further and conduct in vivo validation using mouse models to study the functional impact of the spliced isoform.

Where do you seek scientific inspiration?

Trishna Pani: My inspiration stems from childhood interest in science. I am grateful for my mentors—Dr. Ujjaini Dasgupta and Dr. Avinash Bajaj—who have guided and inspired me with their dedication and passion.
Kajal Rajput: Despite medical advances, cancer remains a challenge. My supervisors inspire me to contribute to solving such complex problems through impactful research.
Animesh Kar: I’m fascinated by discovering new things and solving scientific mysteries. I draw inspiration from Neil deGrasse Tyson and both my Ph.D. supervisors.

How do you intend to help Indian science improve?

While India is making significant strides in research, cancer remains a pressing health issue. As researchers, we aim to contribute to the development of safer and targeted therapies using our understanding of cancer biology and molecular mechanisms.