1. Volume 46, Issue 2, Pages 197-204 (February 2017)
Loss of PTEN Is Associated with Resistance to Anti-PD-1 Checkpoint Blockade Therapy in Metastatic Uterine Leiomyosarcoma 
Suzanne George, Diana Miao, George D. Demetri, Dennis Adeegbe, Scott J. Rodig, Sachet Shukla, Mikel Lipschitz, Ali Amin-Mansour, Chandrajit P. Raut, Scott L. Carter, Peter Hammerman, Gordon J. Freeman, Catherine J. Wu, Patrick A. Ott, Kwok-Kin Wong, Eliezer M. Van Allen 
Immunity 
Volume 46, Issue 2, Pages (February 2017)
DOI: /j.immuni
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2. Immunity 2017 46, 197-204DOI: (10.1016/j.immuni.2017.02.001)
Copyright © 2017 Elsevier Inc. Terms and Conditions

3. Figure 1
Histologic and Radiographic Findings in a Treatment-Naive Patient with Metastatic Uterine Leiomyosarcoma Receiving Anti-PD-1 Monotherapy
(A) Clinical course and tissue collection for immunohistochemical assessment and WES and whole-transcriptome sequencing.
(B) Computed tomography imaging of treatment-responsive tumors and the sole treatment-resistant lesion.
(C) Immunohistochemical staining of the primary and treatment-resistant tumors for PD-1, PD-L1, and PD-L2.
(D) Quantification from representative tumor sections shows a decrease in PD-1+ cell infiltration in the treatment-resistant lesion (p = 0.039, Student’s t test). Error bars represent standard error above and below the mean. ∗p < 0.05.
Immunity  , DOI: ( /j.immuni )
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4. Figure 2
Pre- and Post-treatment Exomic Features in Uterine Leiomyosarcoma in Comparison with TCGA
(A) Computational workflow for WES and whole-transcriptome analysis and neoantigen prediction.
(B) Pre-treatment and resistant tumors had similar neoantigen and mutational loads. See also Table S1.
(C) Integrated Genomics Viewer (IGV_2.3.57) (Thorvaldsdóttir et al., 2013) and lollipop plots showing the TP53 S166∗ and PTEN T131N mutations in pre-treatment and treatment-resistant tumors.
(D) Comparison of the estimated proportion of cancer cells harboring specific mutations in the pre-treatment (x axis) and resistant (y axis) biopsy samples. Shared clonal mutations are shown in the upper right (gray), mutations exclusive to the pre-treatment tumor are in the lower right (blue), and mutations exclusive to the treatment-resistant tumor are in the upper left (red). Lighter shading indicates mutations for which cancer cell fraction distributions are highly uncertain.
(E) Copy-number plots showing amplification of chromosome 8 and a chromosome 10 deletion affecting MYC and PTEN, respectively, in both tumors. See also Figure S1.
(F) Expression of genes related to JAK-STAT and immune inhibitory signaling in untreated sarcoma tumors from the TCGA by PTEN (top) and MYC (bottom) mutational and copy-number status. Boxplot whiskers extend 1.5× the interquartile range from the first and third quartiles. Data beyond the whiskers are outlying points that are plotted individually. ∗p < 0.05, ∗∗p <
Immunity  , DOI: ( /j.immuni )
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5. Figure 3
Predicted and Expressed Neoantigens in Pre-treatment and Treatment-Resistant Tumors
(A) Workflow of neoantigen analysis, including a Venn diagram showing prioritization of putatively response-associated neoantigens.
(B) Line plot showing expression of genes containing predicted neoantigens over time. Each line represents a unique neoantigen; those synthesized for in vitro testing are in green, neoantigens in MB21D2 and QKI are in dark green, and all others are in gray.
(C) IGV showing mRNA transcripts of mutations generating neoantigens in QKI and MB21D2 in the pre-treatment and treatment-resistant tumors.
See also Figure S1, Figure S2, and Table S2.
Immunity  , DOI: ( /j.immuni )
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6. Figure 4
In Vitro Validation of Patient T Cell Reactivity to Predicted Immunogenic Tumor-Specific Peptides
(A) Representative CD4+ versus CD8+ dot plot from viable CD3+ T cells after cell culture.
(B) Representative dot plots showing proportions of interferon-γ-producing CD8+ T cells after peptide incubation.
(C) Bar chart showing proportions of interferon-γ-secreting CD8+ T cells after incubation with tumor-specific mutant peptides (dark orange) or corresponding wild-type control peptides (light orange). Error bars show standard error above and below the mean. ∗p < 0.05, ∗∗p <
(D) Comparison of the reactivity of healthy donor PBMCs (light blue) and patient PBMCs (dark blue) to tumor-specific mutant peptides.
See also Experimental Procedures, Supplemental Information, and Figure S4.
Immunity  , DOI: ( /j.immuni )
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