We describe a unique patient-derived xenograft (PDX) and cell culture model of succinate dehydrogenase-deficient gastrointestinal stromal tumor (SDH-deficient GIST), a rare mesenchymal tumor that can occur in association with paragangliomas in hereditary and non-hereditary syndromes. This model is potentially important for what it might reveal specifically pertinent to this rare tumor type and, more broadly, to other types of SDH-deficient tumors. The primary tumor and xenografts show a very high proliferative fraction, and distinctive morphology characterized by tiny cells with marked autophagic activity. It is likely that these characteristics resulted from the combination of the germline SDHB mutation and a somatic KRAS G12D mutation. The most broadly relevant findings to date concern oxygen and oxidative stress. In paragangliomas harboring SDHx mutations, both hypoxic signaling and oxidative stress are putative drivers of tumor growth. However, there are no models for SDH-deficient paragangliomas. This related model is the first from a SDHB-mutated human tumor that can be experimentally manipulated to study mechanisms of oxygen effects and novel treatment strategies. Our data suggest that tumor growth and survival require a balance between protective effects of hypoxic signaling vs deleterious effects of oxidative stress. While reduced oxygen concentration promotes tumor cell survival, a further survival benefit is achieved with antioxidants. This suggests potential use of drugs that increase oxidative stress as novel therapies. In addition, autophagy, which has not been reported as a major finding in any type of SDH-deficient tumor, is a potential target of agents that might trigger autophagic cell death.
Supplementary Figure.1 Histologic sections of the primary gastric tumor .A. Relatively well differentiated area of tumor showing loosely cohesive epithelioid cells and a minor component of spindle cells (at center). B. Poorly differentiated area of tumor showing numerous rounded tumor cells in a myxoid matrix. There is a gradient of cell size and appearance, ranging from epithelioid cells with conspicuous cytoplasm as in panel A to tiny round cells, some smaller than 10 um, with almost undetectable cytoplasm. Bar = 50 um
Supplementary Figure. 2 High magnification of epithelioid tumor cells showing marked pleomorphism and numerous mitoses (arrows). Bar = 50 um
Supplementary Figure. 3 Abdominal tumor deposit, consisting almost entirely of small rounded cells (A) and prominent pools of myxoid material (B). Bar = 50 um.
Supplementary Figure.4 DOG1 expression in the primary tumor. Staining is less overall than for KIT (cf. Fig. 1), and is absent in most of the small cells. Bar = 50 um
Supplementary Figure 5 representative 13C NMR spectra of Ian GIST (Panel A) compared to control SDH-intact GIST-T1 (Panel B) xenografts.. Select resonance multiplets are labeled with the corresponding molecule and carbon using IUPAC numbering.
Supplementary Table 1. Antibodies for Immunohistochemistry and Immunoblots
Supplementary Table 2. Medium Supplements
Supplementary Table 3 Complete list of RNA-Sequencing analysis of gene expression changed by ≥ tenfold between xenograft and cells cultured in 10%O2