NF2 (neurofibromatosis type 2)-related schwannomatosis (NF2-SWN) is a cancer predisposition syndrome characterized by the development of bilateral vestibular (VS) and spinal schwannomas. While benign, these tumors can cause substantial morbidity, and effective pharmacological treatments remain limited. Here, we demonstrate that genetic ablation of focal adhesion kinase (Fak/Ptk2) impairs tumor formation and preserves hearing in a murine model of NF2. Mechanistically, we show that Fak deletion decreases macrophage infiltration, attenuates nucleotide-binding oligomerization domain-containing protein 2-, leucine rich repeats (LRR)- and pyrin domain-containing protein 3 inflammasome activation, and suppresses the hepatocyte growth factor-MET axis. Pharmacological inhibition of FAK with single agent VS-4718 did not significantly reduce macroscopic tumor volume; however, its use in combination with the mitogen-activated protein kinase kinase (MEK) inhibitor selumetinib resulted in both a significant reduction in tumor volume and the preservation of dorsal root ganglion architecture. Our findings establish a critical role for FAK in schwannoma development and provide rationale for evaluation of combination FAK plus MEK inhibition in future clinical trials for NF2-associated SWN.

Table S13: Enrichment of binding sites for NF-kB and p53 in chromatin regions that become accessible or closed, respectively, upon TONSL overexpression. Genes names and chromatin positions are indicated.

Table S2: Gene expression differences between mature luminal and luminal progenitor cells of the breast. Data are from 15 samples, five each from women of European, African and Latina ancestry.

TCGA MPNST vs GTEX Normal Nerve DEGS

Contains additional materials and methods and supplementary figures

Table S8: Multivariable analysis for overall (N=469) and progression-free (N=453) survival NOT including HER2. Overall and Progression-Free survival data of patients with ER+ breast cancer and treated with endocrine therapy are also shown.

In vitro experimentation in MPNST cell lines

Table S1: Gene expression differences between primary and hTERT-immortalized breast epithelial cell lines.

Table S15: List of genes differentially expressed upon TONSL overexpression in primary cells with genes differentially expressed in TMD-436 upon TONSL knockdown. Fold changes in gene expression are indicated.

Table S5: Description of the patients and characteristics of their tumors (n=597)

TCGA GOBP 2023 Downregulated genes

Table of mice enrolled in drug study

Kaplan Meier Survival Curve and Tolerability

Table S11: The effects of TONSL overexpression on proliferation signature of breast tumors.

Table S9: Results of ATAC-seq of primary and TONSL-immortalized cells.

Table S14: Genes differentially expressed in TMD-436pLKO and TMD-436shTONSL cells. Fold changes in gene expression are indicated.