Reprogramming of somatic cells into organOids: patient-centred neurodevelopmental disease modelling from nascent induced pluripotency

Introduction

Understanding the neurodevelopmental causes and mechanisms of brain disease is challenging due to the limited access to human brain tissues, the complexity of brain anatomy, and the lack of accurate models to recapitulate human brain development.

Challenges in Current Research

The convergence between induced pluripotent stem cell and organoid technology enables the recapitulation of key features of early human neurodevelopmental disease in vitro. However, the following challenges persist:

1. Low efficiency of generating induced pluripotent stem cells (hiPSC) and organoids.
2. High cost associated with the processes.
3. Technical variability that complicates study design.

These challenges make it nearly impossible to design a powered study to associate patient genotype with organoids’ in vitro phenotypes.

Proposed Solution

I will generate technology to produce human brain organoids from selected patients in a robust, timely, and seamless process. This approach aims to capture previously undetected neurodevelopmental phenotypes.

Methodology

This will be achieved by combining reprogramming to nascent states of pluripotency with neurodevelopmental morphogenesis in three dimensions. Starting from a single nascent naive hiPSC, we will develop a process to form:

1. An epiblast cyst.
2. A neuroepithelial cyst.
3. A forebrain organoid in a continuous three-dimensional process.

Application in Fragile X Syndrome

I will use this technology to investigate genetic and epigenetic modifications in the early phase of neural development of Fragile X Syndrome (FXS). We will generate brain organoids from large cohorts of FXS patients for:

i) Profiling the whole spectrum of in vitro phenotypes associated with the variety of patients’ genetic and epigenetic modifications.
ii) Revealing unexplored developmentally regulated mechanisms.
iii) Designing a powered study to assess targeted therapeutic options in FXS.

Impact of ReprOids

ReprOids will allow the generation of human brain organoids from large cohorts of patients, capturing in vitro the whole spectrum of disease manifestations. This will have a huge impact on patient clinical management at the diagnostic, prognostic, or therapeutic level.