RNA binding proteins regulate immune responses
Lead Research Organisation:
University of Cambridge
Department Name: Plant Sciences
Abstract
Theme: Bioscience for Health
The germinal centre (GC) within lymph nodes is a site of intense proliferation by lymphocytes following infection. Upon entry into the GC B lymphocytes acquire an apoptosis sensitive phenotype, undergo immunoglobulin class switching, somatic mutation and selection for high affinity antibody. When B cells exit the GC they may differentiate to become antibody secreting plasma cells or memory B cells, which are typically long-lived cells. At present the molecular mechanisms which regulate this lineage choice are the subject of intensive investigation because of their importance for vaccination and also when these mechanisms are corrupted B cells can undergo transformation to lymphoma and myeloma.
The molecular regulation of proliferation and differentiation requires dynamic coordination of signal transduction pathways with transcriptional and post-transcriptional control of RNA. Our work and that of others shows that this principle underpins the biology of activated B cells in the GC response.
A principal mechanism for regulating RNA abundance and function is through its interaction with specific proteins. The student will test the role of sequence specific RNA binding proteins in B cell activation and the GC response by using a combination of mouse genetics and molecular biology including high throughput sequencing and bioinformatics.
We have evidence that the RNA binding proteins are essential in the GC response. We propose they mediate the rapid removal of mRNA encoding lineage specific transcripts promoting class switch recombination, differentiation into the GC cell and facilitating somatic hyper-mutation. In this study we propose to test the hypothesis that specific RNA binding proteins mediate this function in B cells. We will specifically mutate the genes in B lymphocytes in vivo using a conditional gene targeting approach. The development and function of B cells will then be measured in both naïve and immunized mice. Conditional targeting will be achieved using mb1Cre (B cell specific) and AIDCre (expressed only in activated B cells).
The biology of RNA binding proteins is fast emerging as a major layer of control in all biological processes, but the field lags considerably behind the field of transcription factors. Thus, there is an excellent opportunity for novel discovery relevant to basic bioscience. There is also the potential to discover new processes that contribute to the development of lymphoma and myeloma.
This work will involve training in the use of state of the art conditional gene expression systems in vivo using mouse models; cell biology: molecular biology and bioinformatics.
The germinal centre (GC) within lymph nodes is a site of intense proliferation by lymphocytes following infection. Upon entry into the GC B lymphocytes acquire an apoptosis sensitive phenotype, undergo immunoglobulin class switching, somatic mutation and selection for high affinity antibody. When B cells exit the GC they may differentiate to become antibody secreting plasma cells or memory B cells, which are typically long-lived cells. At present the molecular mechanisms which regulate this lineage choice are the subject of intensive investigation because of their importance for vaccination and also when these mechanisms are corrupted B cells can undergo transformation to lymphoma and myeloma.
The molecular regulation of proliferation and differentiation requires dynamic coordination of signal transduction pathways with transcriptional and post-transcriptional control of RNA. Our work and that of others shows that this principle underpins the biology of activated B cells in the GC response.
A principal mechanism for regulating RNA abundance and function is through its interaction with specific proteins. The student will test the role of sequence specific RNA binding proteins in B cell activation and the GC response by using a combination of mouse genetics and molecular biology including high throughput sequencing and bioinformatics.
We have evidence that the RNA binding proteins are essential in the GC response. We propose they mediate the rapid removal of mRNA encoding lineage specific transcripts promoting class switch recombination, differentiation into the GC cell and facilitating somatic hyper-mutation. In this study we propose to test the hypothesis that specific RNA binding proteins mediate this function in B cells. We will specifically mutate the genes in B lymphocytes in vivo using a conditional gene targeting approach. The development and function of B cells will then be measured in both naïve and immunized mice. Conditional targeting will be achieved using mb1Cre (B cell specific) and AIDCre (expressed only in activated B cells).
The biology of RNA binding proteins is fast emerging as a major layer of control in all biological processes, but the field lags considerably behind the field of transcription factors. Thus, there is an excellent opportunity for novel discovery relevant to basic bioscience. There is also the potential to discover new processes that contribute to the development of lymphoma and myeloma.
This work will involve training in the use of state of the art conditional gene expression systems in vivo using mouse models; cell biology: molecular biology and bioinformatics.
Publications
Description | Proteins which can bind RNA are vital for the mounting of an adaptive immune response by regulating their development and cell killing capacity. |
Exploitation Route | Gain a further understanding as to how post-transciptional regulation of gene expression is a vital component of T lymphocyte function, and that when it is disrupted cells of the immune system fail to mount appropriate immune responses. |
Sectors | Healthcare Pharmaceuticals and Medical Biotechnology |
Title | In vitro cell killing assay |
Description | A method to determine the frequency of target cell killing in vitro using transgenic CD8 T cells and flow cytometry as a means of detection/ |
Type Of Material | Model of mechanisms or symptoms - in vitro |
Year Produced | 2019 |
Provided To Others? | No |
Impact | Using this method i was able to functionally characterise cells which i was using as an in vitro model for CD8 T cell differentiation and development. |
Description | Collaboration with Peter D. Katsikis MD, PhD and the department of immunology at the Erasmus MC |
Organisation | Erasmus MC |
Country | Netherlands |
Sector | Hospitals |
PI Contribution | Supplying materials for in vivo assays of tumour infiltration, and presenting my own findings and exchange of ideas. |
Collaborator Contribution | Performing experiments and exchange of ideas. |
Impact | Partnership is ongoing and experiments are still to be conducted. |
Start Year | 2019 |
Description | Schools visits |
Form Of Engagement Activity | Participation in an open day or visit at my research institution |
Part Of Official Scheme? | No |
Geographic Reach | Regional |
Primary Audience | Schools |
Results and Impact | Performing simple experiments to demonstrate the work which is done by our lab, demonstrations were done with students alone (Key stage 3, GCSE and A-level) and with teachers. Teachers have been in contact since requesting further details on experiments to continue dialogue in the classrooms. |
Year(s) Of Engagement Activity | 2018,2019,2020 |
URL | https://www.babraham.ac.uk/news/2019/11/schools-day-26th-february-2020 |