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

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Contents

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.
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