Step 8

Summary

    My experimental results supported my hypothesis.  Adding the recombinant human interleukin factors IL-3 and IL-6 had profound effects for reprogramming cell culture morphology and growth characteristics.  I achieved my purpose of investigating potential media formulations for HMC-1 cell culture growth in a controlled environment by finding one that led to ideal cell growth patterns.  Using 20 ng/ml IL-3 and 50 ng/ml IL-6 allows for ideal cell culture growth without serum.  This highly controlled culture environment also allows for the precise study of exosomal transfer.

    Important obstacles and unplanned events made completing the research project a challenging process.  A flask of the established human cell line HMC-1.1 was a fortunate gift from Dr. J Butterfield of the Mayo Clinic.  Not much later, a humidity failure in the incubator necessitated full-volume media changes to the first two flasks made in the project, salvaging only the adherent cells.  This made future efforts challenging, but enough cells survived to permit splitting of the flasks into experimental and control groups.  The duration of many of the cell culture procedures also posed a problem, for I stayed at the lab some nights until nearly 11pm.

    During my research, I developed my patent pending ideas for my conception of what the exosomal vector could become.  Though no method has described a self-sufficient intercellular DNA transfer system based on exosomes, the following combined system may serve as a blueprint for realizing this objective.  First, the T7 RNA polymerase autogene system is capable of sustaining cytosolic transcription in various eukaryotic cell lines.  Also, the RepA protein can bind to the DNA from which it is expressed and also function as part of a fusion protein.  Further, DNA has been coupled to DNA-binding fusion proteins bearing a nuclear localization sequence to target their presence in the nucleus.  Also, ceramide has been described as a major factor in targeting to exosomes, and ceramide-binding proteins has been identified.  The unification of these various systems may bring about a new exosomal vector capable of targeting localization to the nucleus for transgene delivery to a cell or to exosomes for delivery to other cells through localization sequences on fusion proteins that the genetic construct is capable of displaying. This would be somewhat similar to an existing system of genetic information transfer through exosomes and microvesicles yet would provide a new level of control over the process. Ultimately, development of such a self-iterating exosomal vector system may enable genetic modification or gene therapy of multicellular systems through its ability to spread through a multicellular system.  Accomplishing this end would revolutionize the world of genetic medicine by enabling individuals to replace or repair a gene in nearly every cell of their bodies.  This would also raise ethical questions regarding the choice to change one's DNA at will for recreational, expressive, or enhancement purposes.  With my promising research on a candidate cell line for the development of the exosomal vector, I hope to be a leading scientist involved in unlocking the potential of exosomes.

Patent Application:

(2017 update: The latest version of my Self-Iterating Exosomal Vector (SIEV) patent application was submitted as the US patent application #15/351,171. This SIEV patent application has the publication number US 20170058296 A1.)