Matt Gethers/20.380 HIV Project/Technical Paper/Matt Design

=Matt Design=

Stem Cell Collection

 * How to collect/purify RBC progenitors?
 * Are there easy kit protocols to do this?
 * What are concentrations of progenitors in blood?
 * How many cells do you need to engineer?
 * How do you store blood and cells during the engineering?
 * Will samples be engineered in the field or sent back to lab facilities?
 * Keeping samples for later use/creation of a library?
 * Need storage facilities and permission from patients.
 * Could these collected progenitors be infected?
 * Possible to cure them?

Progenitor Engineering

 * Need to express CD4+ and Chemokine co-receptors on surface
 * Maybe express library of receptors, or create library of blood cells with different receptors.
 * What will genetic construct look like? Separate operons for everything? Same operon? Polycistronic? Post-transcriptional/translational regulation to further refine time expression profile?
 * What's vector?
 * How transfecting?
 * Need to express RNAses
 * Post-transcriptional/translational regulation to refine expression profile?
 * Protection of progenitor from infection through siRNA.

Reintroduction of Progenitors to Patient

 * Paper to demonstrate that reintroduction of ex vivo cells works.
 * How many cells need to be introduced to the patient in order to establish necessary presence?
 * How to deliver?
 * Injection? Into where?
 * Keep samples of the engineered cells in labs?
 * Need infrastructure and permission from patients.
 * Expand and differentiate cells ex vivo to test before introducing to patient?
 * Need to test every batch of cells before giving to patient.
 * Need FDA blanket approval?

Differentiation in vivo

 * When/how will progenitors be induced to differentiate?
 * Expression profile of genes
 * Modeling of transcription specifically with respect to RNAse expression
 * Need alternate regulation mechanisms to refine time expression profile?
 * Differentiation kinetics, resultant population of progenitors, sink-RBCs.

Therapeutic Action

 * Virions bind RBCs, get caught in trap.
 * What ratio of RBCs to T-cells/virions is necessary for viral ablation?
 * Model birth, death, infection of RBCs, T-cells, HIV.
 * Is appropriate metric surface area and receptor density?
 * Concerns about selecting for anti-trap HIV?
 * Possible to have RBCs evolve with virus through some sort of progenitor sensor system?
 * Drug supersensitivity?

IP

 * Dying, take it away.