| MycoSynVac

Impact on Synthetic Biology

Impact on synthetic biology

The expected impacts on synthetic biology are related to our work on:

Developing a Whole-cell mode

Impact on reinforcing cooperation of industry with academia

The work that should be carried out during the project requires a strong collaboration between industrial and academic leaders. This will lead to valuable strategic links that will aid translational research and its industrial development.

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Environmental and social impacts

An effective vaccine for livestock will increase animal welfare, decrease disease management expenses and reduce the environmental footprint of food production.

Mycoplasma infection in animals cause pain and can lead...

Subprojects

Optimization of Large-Scale Production

A hallmark of the synthetic biology is the model-driven development of circuits, chassis and processes. We will link and embed the dynamic model of central metabolism into a whole-cell constraint-based framework. By incorporating the dynamic metabolic model, we will be able to study the effects of synthetic gene circuits designed to increase growth of M. pneumoniae. Furthermore, we will use the model to identify and test new medium components that could enhance growth rate. The model will be experimentally validated in dedicated bioreactor growth experiments under defined sets of conditions. Measurement of well-chose fluxes will allow capturing the main metabolic features. We will use the genome-scale metabolic models developed to algorithmically generate minimal media “candidate” formulations. We will deploy a series of constraint-based analysis methods to systematically query and ascertain possible flux distributions and genetic construction that can possibly lead to improved growth rates without loss of the other desired functions. The ranked configurations will be assessed under a number of criteria for fitness, feasibility, gain, etc. for subsequent implementation.

Chassis Engineering

The main objective of MycoSynVac is to rationally design a non-virulent M. pneumoniae to obtain a universal chassis optimized to grow in a serum-free medium in bioreactors. We have identified main virulence and pathogenicity factors in M. pneumoniae. We will identify other putative pathogenic and virulence factors by comparing the essentiality of all M. pneumoniae genes with the sequence variability of 22 clinical isolates of M. pneumoniae. These genes will be validated by cell infection assays. M. pneumoniae has an 8-hour doubling time similar to that of other species used for vaccination. However, there are other Mycoplasma species that divide faster. Decreasing the doubling time will significantly improve the industrial efficiency per fermentor. We will follow three approaches to increase growth rate.

Vaccine Design

We will modify existing methods of genome transplantation used for Mycoplasma species. The major challenge in the transplantation procedure is to avoid the destruction of the incoming genome by the specific endonucleases expressed in the recipient cell. Although in MycoSynVac we will work only with strains whose genome has been sequenced, we will develop a pipeline that could be used for any new strain or target species. Adhesins play a crucial role in the primary steps employed by Mycoplasmas while interacting with their host eukaryotic cells using specific mammalian membrane receptors. The physical association of Mycoplasmas with the host cell surface is the basis for the development and persistence of disease, as well as for triggering an immune response. We will do a genome comparison analysis of available Mycoplasma species to identify all putative adhesin genes, and then select those from two target Mycoplasma species. We will replace the main M. pneumoniae adhesins by the counterparts from the three species and test the adhesion and infection properties in in vitro cell culture and/or tracheal assays. Using the genome engineering tools, we will clone and surface-express the selected chimaeric proteins and adjuvants in the chassis. We will then check by western and immunofluorescence if these are recognized by the serum of infected animals.

Exploitation

We will analyze how to bring the product to market after efficacy of the vaccine vector has been shown in studies. We will follow a strict plan regarding steps and dependencies including RACI (Responsible, Accountable, Consulted, Informed) and SIPOC (Suppliers, Inputs, Process, Output and Customers) of the whole process, going from idea/lead to product. There are quite a number of unmet needs from a Mycoplasma vaccine perspective. The first is to have an effective vaccine against mastitis. Specifically, these should be against M. bovis, the most common cause of Mycoplasma mastitis and estimated to be responsible for about 50% or more of the cases of mastitis caused by mycoplasma, as well as against M. bovigenitalium, M. canadense, M. californicum and M. alkalescens, which also affect cattle. Another gap is an efficacious vaccine against contagious agalactia, caused by M. agalactiae, a disease of sheep and goats that is characterized by mastitis, arthritis and keratoconjunctivitis and with devastating effect on the Mediterranean sheep and goat dairy industry. In addition to vaccines, the chassis generated here could have other uses, such as for instance to generate a delivery system for therapeutical applications (e.g. in cell reprogramming, cell therapy, for antibiotics, etc.).

Responsible Research and Innovation

In MycoSynVac we aim to carry out the research and technical development in a responsible manner, following the concept of RRI (Responsible Research and Innovation). To do so we have two partner institutions on board who focus particularly on potential bioethical issues, organize a series of focus groups and Science Cafes with members of the public, hold interviews with relevant stakeholders, produce adequate science communication material (short documentary film, a science game, high school teaching package etc) and prepare material for later risk assessment of the intended products. The feedback we got from our numerous public dialogue events have been presented and discussed within the project consortium to raise awareness within the scientific community about how MycoSynvac is seen by citizens and consumers.