CoPIs: Jörg Jores and Vishvanath Nene [International Livestock Research Institute (ILRI), Nairobi, Kenya]; Carole Lartigue, Alain Blanchard, and Pascal Sirand-Pugnet [National Institute for Agronomy Research (INRA), Bordeaux, France]
Senior Personnel: John Glass (JCVI)
Contagious bovine pleuropneumonia (CBPP), caused by Mycoplasma mycoides subspecies mycoides (Mmm), is an economically very important livestock disease within Africa that limits the availability of protein sources for nutrition and restricts trade. The current control relies on a live vaccine that has shortcomings, such as, limited efficacy and severe side effects. International efforts to create a rational vaccine to protect against CBPP have been thwarted because of an almost complete lack of genetic tools for this species. As a result, it has been difficult to apply the tools of modern biology to this pressing African need. In this project, the International Livestock Research Institute (ILRI), the National Institute for Agronomical Research (INRA) and the J. Craig Venter Institute (JCVI) join forces to explore the use of new synthetic biology technologies to genetically manipulate Mmm genomes to identify virulence traits with the ultimate goal of creating strains that can be developed as live vaccine derivatives. These JCVI synthetic biology approaches are a transformational advance in the ability to design and engineer novel bacteria with extraordinary properties to serve human needs. The JCVI work was done using a closely related species to Mmm, Mycoplasma mycoides subsp. capri (Mmc). During the course of this study, JCVI will continue its work with Mmc, INRA will develop the synthetic biology tools for Mmm as well as the pneumonia-causing strain of Mmc and ILRI will implement the tools and test mutants using in vitro and in vivo assays. While the long term perspective is to use key synthetic biology technologies to produce a safe and efficient Mmm vaccine strain by rationally modifying its genome, the current research is meant to lay the foundation for the development of a modern CBPP vaccine. Several approaches will be pursued. First, the pan genome of the mycoides cluster will be characterized to identify target virulence genes for subsequent mutagenesis. Second, the JCVI synthetic biology tools will be adapted to Mmm at INRA and the synthetic biology technology will be transferred to ILRI in Africa. Third, a caprine model for pulmonary mycoplasma infections will be established with the available tools using the closely related pathogen Mmc. Lastly, the mycoplasma toolbox will be expanded using Mmc as a model in order to enhance the capacity to produce modern Mmm vaccines. The consequences of success will be dramatic and pervasive as not only a novel approach to making vaccines against infectious diseases caused by mycoides cluster members will have been demonstrated but newly developed technical methods will have been transferred to a major research institution in Africa that is capable of sustaining and improving upon what JCVI and INRA seek to accomplish.
This research project will have a major impact on livestock farming in Africa where CBPP is a significant problem. The methods used here will enable the development of vaccine candidates that could not be created previously for this disease. This study also has the potential to revolutionize the area of mycoplasma biology because the methods discussed here will be applied to other members of mycoides cluster and may be applied to mycoplasma pathogens of humans, animals and plants for which there are no tractable genetic methods. An important component involves training of postdoctoral fellows, graduate students as well as undergraduate students as summer interns in the foundations of molecular research. The students will gain valuable insight to cutting-edge research areas such as the new discipline of synthetic biology. The research will also help capacity building of a research institute in a developing country and provide scientific training to an underrepresented segment of society. Information from this study will in the long term benefit the livestock-dependent people by enabling them to increase their protein source from cattle and by reducing restrictions on the trade of cattle and bovine products. The collaborating institutions recognize the need to make the biological resources available to the scientific community. Genome sequencing data of the mycoides cluster will be deposited as rapidly as possible in publically accessible databases such as Genbank (www.ncbi.nlm.nih.gov/genbank/). All biological resources developed under this project will be made available to outside researchers through ILRI and INRA who have agreed to serve as repositories for the strains and to review all requests for materials upon receipt.
