Reduction of sporadic and neurofibromatosis type 2–associated vestibular schwannoma growth in vitro and in vivo after treatment with the c-Jun N-terminal kinase inhibitor AS602801

Abstract

OBJECTIVE Vestibular schwannomas (VSs) are benign nerve sheath tumors that result from mutation in the tumor suppressor gene NF2, with functional loss of the protein merlin. The authors have previously shown that c-Jun N-terminal kinase (JNK) is constitutively active in human VS cells and plays a central role in their survival by suppressing accumulation of mitochondrial superoxides, implicating JNK inhibitors as a potential systemic treatment for VS. Thus, the authors hypothesized that the adenosine 5ʹ-triphosphate–competitive JNK inhibitor AS602801 would demonstrate antitumor activity in multiple VS models. METHODS Treatment with AS602801 was tested in primary human VS cultures, human VS xenografts, and a genetic mouse model of schwannoma (Postn-Cre;Nf2flox/flox). Primary human VS cell cultures were established from freshly obtained surgical tumor specimens; treatment group media was enriched with AS602801. VS xenograft tumors were established in male athymic nude mice from freshly collected human tumor. Four weeks postimplantation, a pretreatment MRI scan was obtained, followed by 65 days of AS602801 (n = 18) or vehicle control (n = 19) treatment. Posttreatment MRI scans were used to measure final tumor volume. Tumors were then harvested. Finally, Postn-Cre;Nf2flox/flox mice were treated with AS602801 (n = 10) or a vehicle (n = 13) for 65 days. Posttreatment auditory brainstem responses were obtained. Dorsal root ganglia from Postn-Cre;Nf2flox/flox mice were then harvested. In all models, schwannoma identity was confirmed with anti-S100 staining, cell proliferation was measured with the EdU assay, and cell death was measured with terminal deoxynucleotidyl transferase–mediated dUTP nick-end labeling staining. All protocols were approved by the local institutional review board and Institutional Animal Care and Use Committees. RESULTS Treatment with AS602801 decreased cell proliferation and increased apoptosis in primary human VS cultures. The systemic administration of AS602801 in mice with human VS xenografts reduced tumor volume and cell proliferation. Last, the AS602801-treated Postn-Cre;Nf2flox/flox mice demonstrated decreased cell proliferation in glial cells in the dorsal root ganglia. However, AS602801 did not significantly delay hearing loss in Postn-Cre;Nf2flox/flox mice up to 3 months posttreatment. CONCLUSIONS The data suggest that JNK inhibition with AS602801 suppresses growth of sporadic and neurofibromatosis type 2–associated VSs. As such, AS602801 is a potential systemic therapy for VS and warrants further investigation.

Schwannomatosis: a Realm Reborn: year one

“…Apart from Bevacizumab, no additional effective treatment options for patients with SWN have emerged: a recent Phase 0 trial of Everolimus, a mTor inhibitor, in patients with meningioma demonstrated only partial inhibition of the pathway in treated tumors [36]; a phase II trial for Icotinib, an oral epidermal growth factor receptor tyrosine kinase inhibitor, in 10 NF2 -SWN patients reported only one case of radiographic response [37]. Preclinical studies have shown interesting results on the effect of Brigatinib in established NF2- deficient xenograft meningiomas and a genetically engineered murine model of spontaneous NF2 schwannomas (results of a human clinical trial are pending, NCT04374305) [38] and of the c-Jun N-terminal-kinase (JNK) inhibitor in primary human vestibular schwannoma cultures and xenografts, and a genetic mouse model of schwannoma [39]. Promising results of gene replacement therapies in NF2 -SWN have also recently been published, using direct intratumoral injection of an adeno-associated virus vector expressing merlin in a novel human schwannoma model in nude mice, leading to regression of tumors over a 10-week period [40].…”