Periodic Reporting for period 2 - GPSorganoids (Generation of Position Specific organoids to study human neuromuscular system development and disease)
Reporting period: 2023-07-01 to 2024-12-31
The proposed project focuses on using human pluripotent stem cells (hPSCs) to generate advanced 3D organoid models to study the development and disease of the human neuromuscular system. The project aims to achieve three major objectives:
1. Generation of Position Specific (GPS) Organoids: Creating precise 3D models of human trunk development by instructing neuromesodermal progenitors (NMPs) to generate position-specific neuromuscular organoids (NMOs). This will enable the study of spinal cord neurons with precise positional identities and their coordinated development with corresponding muscles.
2. Study of Neuromuscular Diseases using NMOs: Utilizing the developed NMOs to model and study early and late-onset neuromuscular diseases, such as spinal muscular atrophy (SMA) and amyotrophic lateral sclerosis (ALS), to understand disease progression and identify potential drug targets.
3. Reconstruction of Human Corticospinal Tracts: Creating a 3D human in vitro model to study the formation and disruption of corticospinal tracts, which are essential for voluntary movements and often affected in neurological disorders, particularly ALS.
The project addresses the challenge of accurately modeling human neuromuscular development and diseases in vitro, particularly the lack of reliable models that incorporate the interactions between neural and muscle cells. Previous models have often focused on neuronal components in isolation, missing crucial cell types involved in the development of neuromuscular diseases. This project aims to overcome these limitations by developing comprehensive neuromuscular organoids that mirror human trunk development and pathology.
Importance for Society:
Understanding the mechanisms underlying neuromuscular development and diseases is critical for developing effective therapies and drugs. By generating 3D organoids that mimic the human neuromuscular system, this project provides a platform to study disease progression, identify potential drug candidates, and ultimately improve treatments for debilitating neuromuscular disorders, benefiting individuals and society as a whole.
1. Generation of Position Specific (GPS) Organoids: Creating precise 3D models of human trunk development by instructing neuromesodermal progenitors (NMPs) to generate position-specific neuromuscular organoids (NMOs). This will enable the study of spinal cord neurons with precise positional identities and their coordinated development with corresponding muscles.
2. Study of Neuromuscular Diseases using NMOs: Utilizing the developed NMOs to model and study early and late-onset neuromuscular diseases, such as spinal muscular atrophy (SMA) and amyotrophic lateral sclerosis (ALS), to understand disease progression and identify potential drug targets.
3. Reconstruction of Human Corticospinal Tracts: Creating a 3D human in vitro model to study the formation and disruption of corticospinal tracts, which are essential for voluntary movements and often affected in neurological disorders, particularly ALS.
The project addresses the challenge of accurately modeling human neuromuscular development and diseases in vitro, particularly the lack of reliable models that incorporate the interactions between neural and muscle cells. Previous models have often focused on neuronal components in isolation, missing crucial cell types involved in the development of neuromuscular diseases. This project aims to overcome these limitations by developing comprehensive neuromuscular organoids that mirror human trunk development and pathology.
Importance for Society:
Understanding the mechanisms underlying neuromuscular development and diseases is critical for developing effective therapies and drugs. By generating 3D organoids that mimic the human neuromuscular system, this project provides a platform to study disease progression, identify potential drug candidates, and ultimately improve treatments for debilitating neuromuscular disorders, benefiting individuals and society as a whole.
Generation of Position-Specific Neuromuscular Models:
We have successfully generated neuromuscular organoids that correspond to anterior spinal cord levels. These organoids are currently being characterized in a manuscript that is in preparation. Additionally, we have established a sophisticated, self-organizing 2D neuromuscular junction model that corresponds to the brachial spinal cord levels. This work has been recently published by Urzi et al. in Nature Communications (2023).
Advancements in Neuromuscular Disease Modeling:
Significant progress has been made in elucidating the mechanisms underlying neuromuscular diseases. Our data from this reporting period demonstrate the potential of neuromuscular organoids (NMOs) as effective models for studying the pathology observed in human diseases. Specifically, we have utilized induced pluripotent stem cell (iPSC) lines derived from patients with Spinal Muscular Atrophy (SMA) and Amyotrophic Lateral Sclerosis (ALS) to generate NMOs. The spinal muscular atrophy organoids exhibit early phenotypic characteristics that faithfully recapitulate the disease pathology.
Technology Transfer:
- We have filed a patent for the method describing the generation of the 2D self-organizing neuromuscular junction model from induced pluripotent stem cells.
- We have received an ERC Proof of Concept grant to automate the production and miniaturization of neuromuscular organoids. This grant will advance our ability to scale up the technology for broader research and potential clinical applications.
We have successfully generated neuromuscular organoids that correspond to anterior spinal cord levels. These organoids are currently being characterized in a manuscript that is in preparation. Additionally, we have established a sophisticated, self-organizing 2D neuromuscular junction model that corresponds to the brachial spinal cord levels. This work has been recently published by Urzi et al. in Nature Communications (2023).
Advancements in Neuromuscular Disease Modeling:
Significant progress has been made in elucidating the mechanisms underlying neuromuscular diseases. Our data from this reporting period demonstrate the potential of neuromuscular organoids (NMOs) as effective models for studying the pathology observed in human diseases. Specifically, we have utilized induced pluripotent stem cell (iPSC) lines derived from patients with Spinal Muscular Atrophy (SMA) and Amyotrophic Lateral Sclerosis (ALS) to generate NMOs. The spinal muscular atrophy organoids exhibit early phenotypic characteristics that faithfully recapitulate the disease pathology.
Technology Transfer:
- We have filed a patent for the method describing the generation of the 2D self-organizing neuromuscular junction model from induced pluripotent stem cells.
- We have received an ERC Proof of Concept grant to automate the production and miniaturization of neuromuscular organoids. This grant will advance our ability to scale up the technology for broader research and potential clinical applications.
This project is focused on making significant advancements beyond the current state of the art in neuromuscular research.
The anticipated outcomes will substantially deepen our understanding of neuromuscular development and the underlying mechanisms of neuromuscular diseases, enhancing our ability to develop targeted drug therapies.
The anticipated outcomes will substantially deepen our understanding of neuromuscular development and the underlying mechanisms of neuromuscular diseases, enhancing our ability to develop targeted drug therapies.