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Basic Science and Pathogenesis.

Thais Rafael Guimarães1, Jung Eun Park1, Catrina Spruce2

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We optimized a protocol to convert marmoset fibroblasts into induced neurons (iNs), creating a valuable model for Alzheimer's Disease (AD) research. This new method enables studying AD pathogenesis and testing therapies in vitro.

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Area of Science:

  • Neuroscience
  • Primate Models
  • Cellular Reprogramming

Background:

  • Alzheimer's Disease (AD) research is limited by models that don't fully replicate human aging and neuropathology.
  • Common marmosets (Callithrix jacchus) offer a valuable non-human primate (NHP) model due to similarities in aging, genetics, and behavior.
  • Direct conversion of fibroblasts to induced neurons (iNs) bypasses pluripotent stages, preserving age-associated phenotypes crucial for AD modeling.

Purpose of the Study:

  • To develop and optimize a protocol for directly converting marmoset fibroblasts into induced neurons (iNs).
  • To establish a reliable in vitro model for studying Alzheimer's Disease (AD) pathogenesis and normal brain aging.
  • To enable high-throughput drug screening and toxicology assessment for AD interventions.

Main Methods:

  • Direct reprogramming of marmoset fibroblasts into induced neurons (iNs).
  • Characterization of marmoset-derived iNs using cellular assays, biochemical and imaging techniques, and RNA sequencing (RNAseq).
  • Comparative analysis with human fibroblast to iN conversion to validate the protocol.

Main Results:

  • Standard human iN conversion protocols were not directly applicable to marmoset fibroblasts, showing species-specific differences.
  • RNA sequencing revealed key differences guiding protocol modifications, including adjustments to conversion length, media composition, and supplementation.
  • An optimized, high-efficiency protocol was established, yielding marmoset iNs with preserved survival, enhanced maturity, and synaptic function.

Conclusions:

  • A novel, robust protocol for marmoset iN conversion was successfully developed.
  • This platform facilitates minimally invasive cellular mechanism studies and advances AD research.
  • The marmoset iN model supports high-throughput drug screening and toxicological assessments for AD therapeutics.