PMC4810029_01
Computed tomography images of perivascular epithelioid cell tumors (PEComas; left panels) at baseline and (right panels) after treatment with sirolimus. All images are chosen to show the maximal size of the lesions indicated by arrows; other lesions may be out of phase in the two studies. (A) Retroperitoneal PEComa before and after 1 year of treatment. (B) Renal PEComa before and after 9 months of treatment. (C, top) Metastatic uterine PEComa in lung before and after 6 weeks of treatment; some lesions are improved, whereas others remain stable. (C, bottom, left) Before and (right) 3 weeks after treatment with sorafenib.
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F1
Restaging studies performed after 6 weeks of treatment revealed interval reduction in size and central cavitation of most of the pulmonary nodules (Fig 1C, top). Significant cavitation of lung masses developed within 2 weeks (Fig 1C, bottom) with improvement in symptoms and obviation of the need for supplemental oxygen, although bilateral pneumothoraces also developed. TSC2, whereas expression generally was maintained in normal vessel wall vascular smooth muscle cells (Fig 2C). The specificity of the anti-TSC2 antibody was confirmed in TSC2-null and -expressing cell lines (Appendix Fig A1).
PMC4810029_02
Computed tomography images of perivascular epithelioid cell tumors (PEComas; left panels) at baseline and (right panels) after treatment with sirolimus. All images are chosen to show the maximal size of the lesions indicated by arrows; other lesions may be out of phase in the two studies. (A) Retroperitoneal PEComa before and after 1 year of treatment. (B) Renal PEComa before and after 9 months of treatment. (C, top) Metastatic uterine PEComa in lung before and after 6 weeks of treatment; some lesions are improved, whereas others remain stable. (C, bottom, left) Before and (right) 3 weeks after treatment with sorafenib.
nihms404581f1.jpg
PMC4810029_03_nihms404581f1.jpg
F1
Restaging studies performed after 6 weeks of treatment revealed interval reduction in size and central cavitation of most of the pulmonary nodules (Fig 1C, top). Significant cavitation of lung masses developed within 2 weeks (Fig 1C, bottom) with improvement in symptoms and obviation of the need for supplemental oxygen, although bilateral pneumothoraces also developed. TSC2, whereas expression generally was maintained in normal vessel wall vascular smooth muscle cells (Fig 2C). The specificity of the anti-TSC2 antibody was confirmed in TSC2-null and -expressing cell lines (Appendix Fig A1).
PMC4810029_03
Computed tomography images of perivascular epithelioid cell tumors (PEComas; left panels) at baseline and (right panels) after treatment with sirolimus. All images are chosen to show the maximal size of the lesions indicated by arrows; other lesions may be out of phase in the two studies. (A) Retroperitoneal PEComa before and after 1 year of treatment. (B) Renal PEComa before and after 9 months of treatment. (C, top) Metastatic uterine PEComa in lung before and after 6 weeks of treatment; some lesions are improved, whereas others remain stable. (C, bottom, left) Before and (right) 3 weeks after treatment with sorafenib.
nihms404581f1.jpg
PMC4810029_03_nihms404581f1.jpg
F1
Restaging studies performed after 6 weeks of treatment revealed interval reduction in size and central cavitation of most of the pulmonary nodules (Fig 1C, top). Significant cavitation of lung masses developed within 2 weeks (Fig 1C, bottom) with improvement in symptoms and obviation of the need for supplemental oxygen, although bilateral pneumothoraces also developed. TSC2, whereas expression generally was maintained in normal vessel wall vascular smooth muscle cells (Fig 2C). The specificity of the anti-TSC2 antibody was confirmed in TSC2-null and -expressing cell lines (Appendix Fig A1).
Perivascular epithelioid cell tumor (PEComa) histology and immunohistochemical stains. Formalin-fixed paraffin-embedded sections stained with (A) hematoxylin and eosin, (B) antiphospho-S6 antibody, and (C) anti-TSC2 antibody.
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F2
Hematoxylin and eosin-stained tumor sections showed sheets of epithelioid cells with granular eosinophilic cytoplasm, intimately associated with blood vessels, and with frequent mitoses, pleomorphism, and necrosis suggestive of malignant behavior (Fig 2A). S100 (not shown). Specimens from all three patients demonstrated strong, diffuse, cytoplasmic staining for phosphorylated S6 protein (Fig 2B) consistent with activation of mTORC1. TSC2, whereas expression generally was maintained in normal vessel wall vascular smooth muscle cells (Fig 2C). The specificity of the anti-TSC2 antibody was confirmed in TSC2-null and -expressing cell lines (Appendix Fig A1).
Mulitplex ligation-dependent probe amplification (MLPA) and fluorescent in situ hybridization (FISH) assays of TSC1 and TSC2 in perivascular epithelioid cell tumor (PEComa) samples. Genomic DNA from (A-C) tumor or (D) peripheral blood assessed for copy number by mulitplex ligation-dependent probe amplification (MLPA). Signals were normalized to values from control tissues. (E) FISH using (green) TSC1-specific DNA probes, (red) chromosome 16 centromeric probes, and nuclei isolated from uterine PEComa tissue.
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F3
The integrity of the TSC1 and TSC2 genomic loci was examined by MLPA (Fig 3). Genomic DNA was extracted from paraffin-embedded tumor tissue (Figs 3A, 3B, and 3C) or peripheral blood (Fig 3D) and was assessed for copy number changes by MLPA by using nine exonic probes for TSC1 and TSC2, respectively, and five control probes from other genomic locations. Marked reduction in signal from the TSC1 gene was observed in the uterine tumor specimen from patient 3 (Figs 3C to D), consistent with somatic biallelic deletion of TSC1 (Fig 3A). MLPA demonstrated no alterations in tumor samples from the retroperitoneal PEComa (patient 1; Fig 3A) or the renal PEComa (patient 2; Fig 3B), which suggests that mechanisms other than gross alteration of the TSC1 or TSC2 genomic loci were important in loss of TSC2 expression in those two tumors.