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Updated: Nov 19, 2025
Subpial Adeno-associated Virus 9 AAV9 Vector Delivery in Adult Mice
Published on: July 13, 2017
Gene therapy for tuberous sclerosis complex type 2 in a mouse model by delivery of AAV9 encoding a condensed form of
Pike-See Cheah1,2, Shilpa Prabhakar1, David Yellen1
1Molecular Neurogenetics Unit, Department of Neurology and Center for Molecular Imaging Research, Department of Radiology, Massachusetts General Hospital, and Program in Neuroscience, Harvard Medical School, Boston, MA, USA.
Abstract:
Tuberous sclerosis complex (TSC) results from loss of a tumor suppressor gene - TSC1 or TSC2, encoding hamartin and tuberin, respectively. These proteins formed a complex to inhibit mTORC1-mediated cell growth and proliferation. Loss of either protein leads to overgrowth lesions in many vital organs. Gene therapy was evaluated in a mouse model of TSC2 using an adeno-associated virus (AAV) vector carrying the complementary for a "condensed" form of human tuberin (cTuberin). Functionality of cTuberin was verified in culture. A mouse model of TSC2 was generated by AAV-Cre recombinase disruption of Tsc2-floxed alleles at birth, leading to a shortened lifespan (mean 58 days) and brain pathology consistent with TSC. When these mice were injected intravenously on day 21 with AAV9-cTuberin, the mean survival was extended to 462 days with reduction in brain pathology. This demonstrates the potential of treating life-threatening TSC2 lesions with a single intravenous injection of AAV9-cTuberin.
Insights
Gene therapy using adeno-associated virus (AAV) carrying condensed tuberin (cTuberin) shows promise for Tuberous Sclerosis Complex (TSC). A single AAV9-cTuberin injection significantly extended survival and reduced brain pathology in a TSC2 mouse model.
Area of Science:
- Genetics
- Molecular Biology
- Gene Therapy
Background:
- Tuberous Sclerosis Complex (TSC) is a genetic disorder caused by mutations in tumor suppressor genes TSC1 or TSC2.
- Loss of TSC1 (hamartin) or TSC2 (tuberin) proteins disrupts mTORC1 signaling, leading to uncontrolled cell growth and organ lesion formation.
- Current treatments for TSC focus on managing symptoms, with limited options for addressing the underlying genetic cause.
Purpose of the Study:
- To evaluate the efficacy of gene therapy using an adeno-associated virus (AAV) vector delivering a functional form of tuberin (cTuberin) in a mouse model of TSC2.
- To assess the impact of AAV9-cTuberin gene therapy on survival rates and brain pathology in TSC2-deficient mice.
Main Methods:
- A mouse model of TSC2 was established by disrupting the Tsc2 gene using AAV-Cre recombinase, resulting in reduced lifespan and neurological abnormalities.
- Adeno-associated virus serotype 9 (AAV9) carrying a condensed form of human tuberin (AAV9-cTuberin) was administered intravenously to affected mice at 21 days of age.
- Survival and brain pathology were monitored and compared between treated and untreated TSC2 mouse models.
Main Results:
- Mice treated with AAV9-cTuberin exhibited a significant increase in mean survival, extending from 58 days to 462 days.
- Intravenous AAV9-cTuberin gene therapy led to a notable reduction in brain pathology characteristic of TSC.
- In vitro studies confirmed the functionality of the delivered cTuberin protein.
Conclusions:
- A single intravenous injection of AAV9-cTuberin gene therapy holds significant potential for treating TSC2-related lesions.
- This gene therapy approach offers a promising strategy for extending survival and mitigating the severe neurological consequences of Tuberous Sclerosis Complex.
