TARGESON, INC. — Department of Health and Human Services SBIR Phase I: NIAID

Description

DESCRIPTION provided by applicant Products for transfection have long been a staple of biomedical research and many are now finding potential applications in clinical medicine In particular chemical type and electroporation products are widely used owing to their ease of use relative to viral transfection Despite the commercial success in this segment efficient transfection of some cell types remains difficult Many of the cell types that are generally resistant to non viral transfection are immune cells lymphocytes macrophages which places a bottleneck in the development of cellular therapy and basic science In this proposal we seek to validate a unique non viral mechanism of transfection with particular emphasis on cells that existing products do poorly with primary B and T cells Our technology is based on sonoporation in which transient poration is mediated by the interaction of ultrasound energy and compressible microbubble reagents Despite some recent progress demonstrating feasibility of sonoporation for treating difficult cells significant work remains to be done in orer to make this technology broadly useful We have identified performance milestones pertaining to efficiency and purity that are necessary for a viable commercial product We have developed a set of tools that enable high throughput evaluation of multiple procedural parameters concentration acoustic intensity pulse length repetition frequency microbubble diameter treatment duration in a well plate based assay and describe an optimization path designed to find treatment conditions under which high transfection efficiency andgt and cell viability andgt are achieved We plan to develop a product set consisting of a sonoporation device and cell type specific microbubble reagents that enables isolation of desired cells from spleen or blood and subsequent transfection using a rapid and robust procedure We will first evaluate delivery efficiency and viability in a panel of cultured lymphocyte cell lines with the goal of reducing the parameter space before testing on primary cells We will then evaluate transfection efficiency for model RNA and DNA payloads in primary mouse T and B cells We will complete our proof of concept evaluation by investigating the potential for functional alterations in primar cells transfected using our products Success in the aims described here will result in data that enables us to begin prototyping a customer ready sonoporation device and to begin commercialization discussions with potential distribution partners Downstream applications in clinical cell therapy are anticipated to be the topic for a phase II proposal PUBLIC HEALTH RELEVANCE Several techniques for transferring nucleic acid transfection into cells are commercially available although routinely achieving high efficiency transfection without significant cell death is difficult for many cell types This is particularly true for primry lymphocytes B and T cells obtained from humans and mice which have broad relevance in oncology immunology and cardiovascular disease This deficiency slows progress in basic science and hinders the development of clinical cell and gene therapy products The project proposed here will explore the feasibility a novel mechanism of transfection that may overcome the inherent limitations of existing transfection technologies We propose to rigorously evaluate an ultrasound based method known as sonoporation in conjunction with a microparticle reagent that can both isolate desired cells from a patient sample and mediate the transfection The proposed product is a sonoporation device and a line of cell type specific microparticle reagents designed for transfecting difficult cell types Successful completion of this project will result in technology able to mediate high efficiency and low cost gene transfer for use in biomedical research with potential for translation to clinical use