A cerebellopontine angle mouse model for the investigation of tumor biology, hearing, and neurological function in NF2-related vestibular schwannoma

Abstract

Neurofibromatosis type II (NF2) is a disease that lacks effective therapies. NF2 is characterized by bilateral vestibular schwannomas (VSs) that cause progressive and debilitating hearing loss, leading to social isolation and increased rates of depression. A major limitation in NF2 basic and translational research is the lack of animal models that allow the full spectrum of research into the biology and molecular mechanisms of NF2 tumor progression, as well as the effects on neurological function. In this protocol, we describe how to inject schwannoma cells into the mouse brain cerebellopontine angle (CPA) region. We also describe how to apply state-of-the-art intravital imaging and hearing assessment techniques to study tumor growth and hearing loss. In addition, ataxia, angiogenesis, and tumor–stroma interaction assays can be applied, and the model can be used to test the efficacy of novel therapeutic approaches. By studying the disease from every angle, this model offers the potential to unravel the basic biological underpinnings of NF2 and to develop novel therapeutics to control this devastating disease. Our protocol can be adapted to study other diseases within the CPA, including meningiomas, lipomas, vascular malformations, hemangiomas, epidermoid cysts, cerebellar astrocytomas, and metastatic lesions. The entire surgical procedure takes ~45 min per mouse and allows for subsequent longitudinal imaging, as well as neurological and hearing assessment, for up to 2 months.

Combination therapy with mTOR kinase inhibitor and dasatinib as a novel therapeutic strategy for vestibular schwannoma

“…Second, using this model, evaluation of the neurological function is limited to rotarod assay, which only assesses motor coordination and does not reflect sensory function. Future preclinical efficacy studies using the orthotopic models [52][53][54] and NF2 genetically engineered mouse models [45][46][47][48][49][50] will complement the current study and facilitate translation to clinic. Though dasatinib has been associated with some toxicity in the clinic, we did not observe systemic toxicity (body weight loss) in mice treated with AZD2014, dasatinib, or combination therapy.…”

Current Understanding of Neurofibromatosis Type 1, 2, and Schwannomatosis

“…Recently, Chen et al described the injection of schwannoma cells into the mouse brain cerebellopontine angle region and the application intravital imaging and hearing assessment techniques to study tumor growth and hearing loss. In addition, ataxia, angiogenesis, and tumor–stroma interaction assays could be shown [ 107 ].…”

Uncoupling Traditional Functionalities of Metastasis: The Parting of Ways with Real-Time Assays

“…In a mouse model, intra-vital imaging assisted approaches elucidated the role of Rac1-dependent membrane protrusions in the maintenance of 3D spatial positions of mouse dermal fibroblasts during wound healing [140]. In yet another report, the transplantation of tumor cells into the mouse cerebellopontine angle region coupled with intra-vital imaging successfully developed a model to study the disease progression of vestibular schwannoma [141]. Separately, the intra-vital imaging of early breast carcinoma lesions identified a Her2 driven dissemination program that enables the dispersal of dormant cancer cells to distant organs, prior to the development of a primary tumor [142].…”