R R Tubbs

Case Western Reserve University School of Medicine, Cleveland, Ohio, United States

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Publications (419)2221.33 Total impact

  • [Show abstract] [Hide abstract]
    ABSTRACT: We studied the global miRNA expression in diffuse large B-cell lymphoma (DLBCL; n=79), Burkitt lymphoma (BL; n= 36), primary mediastinal B-cell lymphoma (PMBL; n=12), B-cell lines (n=11), and normal subsets of naïve B-cells (N), centroblasts (CB), and peripheral blood B-cells along with their corresponding gene expression profiles (GEP). The normal B-cell subsets have well-defined miRNA signatures. The CB miRNA signature was significantly associated with germinal center B-cell (GCB)-DLBCL compared to ABC-DLBCL (p=0.002). We identified a 27-miRNA signature that included MYC targets and enabled the differentiation of BL from DLBCL, a distinction comparable with the "gold standard" GEP-defined diagnosis. Distinct miRNA signatures were identified for DLBCL subgroups, including GCB-DLBCL, activated B-cell (ABC)-DLBCL and PMBL. Interestingly, most of the unclassifiable-DLBCL by GEP showed a strong similarity to the ABC-DLBCL by miRNA expression profiling. Consistent results for BL and DLBCL subgroup classification were observed in formalin-fixed, paraffin-embedded tissue, making such tests practical for clinical use. We also identified predictive miRNA biomarker signatures in DLBCL, including high expression of miR-155 which significantly associated with R-CHOP response failure. This finding was further supported by the observation that high expression of miR-155 sensitizes cells to AKT inhibitors in vitro, suggesting a novel treatment option for resistant DLBCL. Copyright © 2014 American Society of Hematology.
    Blood 12/2014; DOI:10.1182/blood-2014-04-566778 · 9.78 Impact Factor
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    ABSTRACT: Background Detection of immunoglobulin light-chain restriction is important in the diagnosis of B-cell non-Hodgkin lymphoma (B-NHL). Flow-cytometry, commonly used to evaluate light-chain restriction, is impractical to be used in cutaneous specimens. Immunohistochemical and conventional chromogenic in situ hybridization (CISH) methods on formalin-fixed-paraffin-embedded (FFPE) tissue lack sufficient sensitivity to detect low-level light-chain expression in B-NHL without plasmacytic differentiation. Ultrasensitive bright-field mRNA-ISH (BRISH) for in situ light-chain detection in cutaneous B-NHL has been assessed.DesignKappa/lambda mRNA was detected using two-color BRISH (RNAscope 2xPlex, Advanced Cell Diagnostics) on 27 FFPE skin biopsies and excisions from patients with available B-cell PCR clonality studies: 16 clonal B-cell lesions (6 follicle center lymphoma, 5 marginal zone lymphoma, 3 large B-cell lymphoma, and 2 other) and 11 non-clonal B-cell proliferations.ResultsBRISH was successful in 15/16 clonal B-cell lesions and 11/11 non-clonal proliferations. Light-chain restriction was detected in 15/15 clonal lesions and in 1/11 non-clonal proliferations (96.1% overall concordance with clonality PCR). In 4/5 marginal zone lymphomas, light-chain restriction was detected as strong monotypic mRNA expression in a B-cell subset, consistent with plasmacytic differentiation.Conclusion Ultrasensitive BRISH can successfully detect light-chain restriction in B-NHL from FFPE skin specimens and may be a useful adjunct ancillary tool in cases not resolved by CISH or immunohistochemical methods.
    Journal of Cutaneous Pathology 11/2014; 42(2). DOI:10.1111/cup.12415 · 1.56 Impact Factor
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    ABSTRACT: Patients with aggressive, BCL2 protein-positive (+) diffuse large B-cell lymphoma (DLBCL) often experience rapid disease progression that is refractory to standard therapy. However, there is potential for false-negative staining of BCL2 using the standard monoclonal mouse 124 antibody that hinders the identification of these high-risk DLBCL patients. Herein, we compare two alternative rabbit monoclonal antibodies (E17 and SP66) to the 124 clone in staining for BCL2 in formalin-fixed, paraffin-embedded DLBCL tissues. Overall, in two independent DLBCL cohorts E17 and SP66 detected BCL2 expression more frequently than 124. In the context of MYC expression, cases identified as BCL2 (+) with SP66 demonstrated the strongest correlation with worse OS. The 124 clone failed to detect BCL2 expression in the majority of translocation (+), amplification (+), and activated B-cell DLBCL cases in which high levels of BCL2 protein are expected. Using dual in-situ hybridization (Dual ISH) as a new tool to detect BCL2 translocation and amplification, we observed similar results as previously reported for fluorescence ISH for translocation but a higher amplification frequency, indicating that BCL2 amplification may be under-reported in DLBCL. Among the discrepant cases, phosphorylation of BCL2 at T69 and/or S70 was more common than in the concordant cases and may contribute to the 124 false-negatives, in addition to previously associated mutations within the epitope region. The accurate detection of BCL2 expression is important in the prognosis and treatment of DLBCL particularly with new anti-BCL2 therapies.
    Human pathology 10/2014; 45(10). DOI:10.1016/j.humpath.2014.06.005 · 2.81 Impact Factor
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    ABSTRACT: Epidermal growth factor receptor (EGFR) gene mutation status should be determined in all patients with advanced, non-squamous non-small cell lung carcinoma (NSCLC) to guide targeted therapy with EGFR tyrosine kinase inhibitors. EGFR mutations are commonly tested by Sanger sequencing or allele specific polymerase chain reaction (ASPCR) on formalin-fixed paraffin-embedded (FFPE) samples including cell blocks (CB) that may fail due to absence of tumor cells. The cell pellet from cytology specimens obtained at the time of endobronchial guided ultrasound fine needle aspiration (EBUS FNA) (EBUS-TBNA, transbronchial needle aspiration) represents an alternative resource for additional tissue. Here we demonstrate the utility of using the FNA cell pellet versus for the detection of EGFR mutations in NSCLC.
    Lung cancer (Amsterdam, Netherlands) 09/2014; 86(2). DOI:10.1016/j.lungcan.2014.09.003 · 3.14 Impact Factor
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    ABSTRACT: Aims PCR studies for lymphoid clonality are now widely employed, especially using Euroclonality/BIOMED-2 primers. Criteria for interpretation as a clonal result, however, have proven controversial. This study examines the frequency and clinical significance of equivocal amplification patterns and measures the interobserver reproducibility of clonality interpretations. Methods At our institution, results of each primer set are first classified as clonal, non-clonal or abnormal (equivocal peak on polyclonal background). Final results for all primer sets are then collectively reported as positive (>= 1 clonal result), negative (non-clonal results) or indeterminate (>= 1 abnormal result) for a clonal population. Results of 274 consecutive clonality cases were reviewed, and the interobserver reproducibility of individual primer set reactions and final results was determined in a subset of 30 cases. Results 44/161 (27%) B-cell and 50/163 (31%) T-cell cases contained at least one abnormal peak. Of these, 29 (64%) and 31 (62%), respectively, showed clonal results in another primer set. Interobserver reproducibility was excellent for most primer sets and for final interpretations, but only fair to good for IGK V-J and TCRB D-J1+2 primer sets. A definitive diagnosis of lymphoma was rendered in 93%, 20% and 6% of B-cell cases and 90%, 42%, and 14% of T-cell cases positive, indeterminate or negative for a clonal population, respectively. Conclusions Using a subjective approach, abnormal (equivocal) peaks are frequently observed in routine practice. However, most cases with abnormal peaks contain clonal rearrangements in other primer sets, facilitating overall interpretation of final results with excellent interobserver reproducibility.
    Journal of Clinical Pathology 09/2014; 67(12). DOI:10.1136/jclinpath-2014-202454 · 2.55 Impact Factor
  • Nature 09/2014; · 42.35 Impact Factor
  • Zsuzsanna Varga, Raymond R Tubbs, Holger Moch
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    ABSTRACT: IntroductionHER2 status assessment became a mandatory test assay in breast cancer, giving prognostic and predictive information including eligibility for adjuvant anti-HER2 therapy. Precise and reliable assessment of HER2 status is therefore of utmost importance. In this study we analyzed breast cancer samples by a novel technology for concomitant detection of the HER2 protein and gene copy number.MethodsTissue microarrays containing 589 invasive breast cancer samples were analyzed with a double immunohistochemistry (IHC) and silver labeled in situ hybridization (SISH) assay simultaneously detecting HER2 protein and gene copy number in the same tumor cells. This bright-field assay was analyzed using scores according to the modified ASCO guidelines and the results were correlated with patient prognosis.ResultsOverall concordance rate between protein expression and the presence of gene amplification was 98%. Fifty-seven of 60 tumors (95%) with IHC score 3+, 6 of 10 tumors with IHC score 2+ (60%) and only 3 of 519 tumors (0.6%) with IHC score 0/1+ were amplified by SISH. Patients with gene amplification despite IHC score 0/1+ had a tendency for worse overall survival (p = 0.088, reaching nearly statistical significance) compared to IHC score 0/1+ without amplification. In contrast, there was no difference in overall survival in IHC score 3+/2+ tumors with and without gene amplification.ConclusionsThe novel double IHC and SISH assay for HER2 is efficient in the identification of breast cancer with discordant HER2 protein and HER2 gene status, especially for the prognostically relevant groups of HER2 protein negative tumors with HER2 amplification and HER2 protein positive tumors without HER2 amplification. Breast cancer without HER2 amplification among IHC score 2+/3+ tumors (10% in our cohort) suggests that other mechanisms than gene amplification contribute to protein overexpression in these cells.
    PLoS ONE 08/2014; 9(8):e105961. DOI:10.1371/journal.pone.0105961 · 3.53 Impact Factor
    This article is viewable in ResearchGate's enriched format
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    ABSTRACT: Cytogenetic alterations are strong outcome prognosticators in uveal melanoma (UVM). Monosomy 3 (-3) and MYC-amplification at 8q24 are commonly tested by fluorescence-in-situ-hybridization (FISH). Alternatively, microarray analysis provides whole-genome data, detecting partial chromosome loss, loss-of-heterozygosity (LOH) or abnormalities unrepresented by FISH probes. Non-fixed frozen tissue is conventionally used for microarray analysis but may not always be available. We assessed the feasibility of genomic microarray analysis for high resolution interrogation of UVM using formalin-fixed paraffin-embedded tissue (FFPET) as an alternative to frozen tissue (FZT). Enucleations from 44 patients (clinical trial NCT00952939) yielded sufficient DNA from FFPET (n=34) and/or frozen tissue (n=41) for comparative-genomic-hybridization and select single nucleotide polymorphism-analysis (CGH/SNP) on Roche®-NimbleGen® OncoChip® arrays. CEP3-FISH was performed on matched cytology ThinPrep material. CGH/SNP analysis was successful in 30/34 FFPET and 41/41 FZT samples. 26/27 (96.3%) paired FFPET/FZT samples were concordant for at least 4 of 6 major recurrent abnormalities (-3, +8q, -1p, +6p, -6q, -8p) and 25/27 (92.6%) were concordant for -3. CGH/SNP was concordant with CEP3-FISH in 27/30 (90%) FFPET and 38/41 (92.6%) FZT cases, detecting partial -3q in 2 CEP3-FISH-negative cases and whole-chromosome 3, 4 and 6 SNP-LOH in 1 case. CGH-detection of -3, +8q, -8p on FFPET and FZT showed significant correlation with the clinical outcome measures (metastasis development, time to progression, survival). UVM genotyping by CGH/SNP on FFPET is highly concordant with FZT analysis and with CEP3-FISH and is a practical method for UVM prognostication. Genome-wide coverage provides additional data with potential relevance to UVM biology, diagnosis and prognosis.
    Cancer Genetics 08/2014; DOI:10.1016/j.cancergen.2014.08.005 · 2.42 Impact Factor
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    ABSTRACT: The goal of personalized medicine is to treat patients with a therapy predicted to be efficacious based on the molecular characteristics of the tumor, thereby sparing the patient futile or toxic therapy. Anaplastic lymphoma kinase (ALK) inhibitors are effective against ALK-positive non-small-cell lung cancer (NSCLC) tumors, but to date the only approved companion diagnostic is a break-apart fluorescence in situ hybridization (FISH) assay. Immunohistochemistry (IHC) is a clinically applicable cost-effective test that is sensitive and specific for ALK protein expression. The purpose of this study was to assemble an international team of expert pathologists to evaluate a new automated standardized ALK IHC assay. Archival NSCLC tumor specimens (n =103) previously tested for ALK rearrangement by FISH were provided by the international collaborators. These specimens were stained by IHC with the anti-ALK (D5F3) primary antibody combined with OptiView DAB IHC detection and OptiView amplification (Ventana Medical Systems, Inc., Tucson, AZ). Specimens were scored binarily as positive if strong granular cytoplasmic brown staining was present in tumor cells. IHC results were compared with the FISH results and interevaluator comparisons made. Overall for the 100 evaluable cases the ALK IHC assay was highly sensitive (90%), specific (95%), and accurate relative (93%) to the ALK FISH results. Similar results were observed using a majority score. IHC negativity was scored by seven of seven and six of seven evaluators on three and two FISH-positive cases, respectively. IHC positivity was scored on two FISH-negative cases by seven of seven readers. There was agreement among seven of seven and six of seven readers on 88% and 96% of the cases before review, respectively, and after review there was agreement among seven of seven and six of seven on 95% and 97% of the cases, respectively. On the basis of expert evaluation the ALK IHC test is sensitive, specific, and accurate, and a majority score of multiple readers does not improve these results over an individual reader's score. Excellent inter-reader agreement was observed. These data support the algorithmic use of ALK IHC in the evaluation of NSCLC.
    Journal of thoracic oncology: official publication of the International Association for the Study of Lung Cancer 05/2014; 9(5):631-8. DOI:10.1097/JTO.0000000000000115 · 4.55 Impact Factor
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    ABSTRACT: Overexpression of MET receptor tyrosine kinase and its ligand hepatocyte growth factor (HGF) and MET gene amplification have been well-documented in non-small-cell lung cancer (NSCLC). Activated MET signaling plays an important role in human cancer tumorigenesis, metastasis, and drug resistance. However, the deregulation of MET/HGF pathway in NSCLC harboring ALK gene rearrangement (ALK[+]), which is sensitive to dual ALK and MET inhibitor Crizotinib, has not been reported. We performed systematic analysis of MET/HGF expression by immunohistochemistry (IHC) and MET gene amplification by dual color, dual hapten bright field in situ hybridization in 19 ALK(+) and 73 ALK(-) NSCLC tumor tissues from those who had clinical ALK rearrangement test done at the Cleveland Clinic from August 2010 to January 2013. IHC scoring was interpreted on a standard four-tier system. The percentage of MET IHC score 0, 1+, 2+, and 3+ were 5.5%, 27.8%, 50.0%, and 16.7% in ALK(+) group, compared with 28.8%, 33.9%, 23.7%, and 13.6% in ALK(-) group, respectively. The MET high expression (IHC score 2 or 3) was significantly higher in ALK(+) group statistically (66.7% versus 37.3%, p = 0.03). HGF-high expression (IHC score 2 or 3) was 33.3% in ALK(+) and 15.8% in ALK(-) (p = 0.17). We identified eight cases in ALK(-) and one case in ALK(+) tumor who had MET gene amplification (18.4% versus 7.1%, p = 0.43) by dual color, dual hapten bright field in situ hybridization. No significant correlation between MET protein receptor expression and gene amplification was identified. Our study demonstrated for the first time that MET receptor expression, but not MET gene amplification, is significantly increased in ALK(+) NSCLC. MET gene amplification is a relatively rare event in this unique population compared with ALK(-) NSCLC.
    Journal of thoracic oncology: official publication of the International Association for the Study of Lung Cancer 05/2014; 9(5):646-53. DOI:10.1097/JTO.0000000000000145 · 4.55 Impact Factor
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    ABSTRACT: The clinicopathologic findings in Burkitt lymphoma (BL) and diffuse large B-cell lymphoma (DLBCL) may show significant overlap, and MYC abnormalities, found in all BLs, also occur in a subset of DLBCL. The 2008 World Health Organization classification introduced the category of "B-cell lymphoma, unclassifiable, with features intermediate between DLBCL and BL" (BCLU) in recognition of this overlap, but the clinical significance of BCLU (ie, "high-grade") morphology and the relationship between BCLU morphology and MYC abnormalities remains unclear. In this study, we identified 260 cases of non-Burkitt, diffuse aggressive B-cell lymphomas from SWOG S9704, a phase 3 randomized study of standard immunochemotherapy versus autologous stem cell transplantation. Of these, 31 cases (12%) showed BCLU morphology, and 229 (88%) showed typical DLBCL morphology. Of 198, 27 (14%) were positive for MYC by immunohistochemistry. BCLU morphology was associated with an increased incidence of MYC expression but otherwise was not associated with distinct clinicopathologic features or significantly decreased survival. MYC-positive cases were morphologically and phenotypically heterogenous and were associated with poor progression-free and overall survival in multivariate analysis. These findings confirm that BCLU does not represent a distinct clinicopathologic entity and demonstrate that BCLU morphology alone does not significantly impact survival compared with typical DLBCL. In contrast, MYC protein expression is a poor prognostic factor that may be associated with either BCLU or DLBCL morphology, and MYC immunohistochemistry is suggested for routine prognostic evaluation (Clinicaltrials.gov identifier: NCT00004031).
    The American journal of surgical pathology 04/2014; 38(4):494-501. DOI:10.1097/PAS.0000000000000147 · 4.59 Impact Factor
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    ABSTRACT: Oncogenic anaplastic lymphoma kinase (ALK) gene rearrangements in non-small-cell lung carcinomas (NSCLC) provide the basis for targeted therapy with crizotinib and other specific ALK inhibitors. Treatment eligibility is conventionally determined by the Food and Drug Administration-approved companion diagnostic fluorescence in situ hybridization (FISH) assay on paraffin-embedded tissue (PET). On limited samples such as fine needle aspiration-derived cytoblocks, FISH for ALK is often uninformative. FISH performed on liquid-based ThinPrep slides (ThinPrep-FISH) may represent a robust alternative. Two hundred thirty cytology samples from 217 patients with advanced NSCLC, including a consecutive series of 179 specimens, were used to generate matched ThinPrep slides and paraffin cytoblocks. The same ThinPrep slides used for cytologic diagnosis were assessed by standard ALK break-apart two-color probe FISH, after etching of tumor areas. Ultrasensitive ALK immunohistochemistry (IHC) on corresponding cytoblocks [D5F3 antibody, OptiView signal amplification] served as the reference data set. ThinPrep-FISH ALK signals were robust in 228 of 230 cases and not compromised by nuclear truncation inherent in paraffin-embedded tissue-FISH; only two samples displayed no signals. Nine of 178 informative cases (5%) in the consecutive series and 18 of 228 informative cases (7.8%) overall were ALK rearranged by ThinPrep-FISH. In 154 informative matched ThinPrep-FISH and cytoblock-IHC samples, 152 were concordant (10, 6.5% ALK status positive; 142, 92.2% ALK status negative), and two (1.3%) were ThinPrep-FISH positive but IHC negative (sensitivity 100%, specificity 98.6%, overall agreement 98.7%). Detection of ALK gene rearrangements in liquid cytology ThinPrep slides derived from patients with NSCLC can be confidently used for clinical ALK molecular testing.
    Journal of thoracic oncology: official publication of the International Association for the Study of Lung Cancer 04/2014; 9(4):464-8. DOI:10.1097/JTO.0000000000000104 · 4.55 Impact Factor
  • Cancer Research 03/2014; 73(24 Supplement):P1-02-04-P1-02-04. DOI:10.1158/0008-5472.SABCS13-P1-02-04 · 9.28 Impact Factor
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    ABSTRACT: Peripheral T-cell lymphoma (PTCL) encompasses a heterogeneous group of neoplasms with generally poor clinical-outcome. Currently 50% of PTCLs are not-classifiable (PTCL-NOS).Gene-expression profiles on 372 PTCLs were analyzed and findings were validated by immunohistochemistry and mutation analysis. Robust molecular classifiers and oncogenic pathways that reflect the pathobiology of tumor cells and their microenvironment were identified for major PTCL-entities, including angioimmunoblastic T-cell lymphoma (AITL; n=114), anaplastic lymphoma kinase (ALK)-positive (n=31) and ALK-negative anaplastic large cell lymphoma (n=48), adult T-cell leukemia/lymphoma (n=14) and extranodal NK/T-cell lymphoma (ENKTL). ENKTL were further separated into NK-cell (n=23) and γδT-cell lymphomas (n=21). We re-classified 37% of morphologically-diagnosed PTCL-NOS cases into other specific subtypes by molecular signatures. Pathologic re-examination, immunohistochemistry and IDH2 mutation analysis supported the validity of re-classification. The remaining PTCL-NOS cases (n=121) were classified into two major molecular subgroups, characterized by high expression of either GATA3 (33%;40/121) or TBX21 (49%;59/121) and corresponding target genes. GATA3-subgroup was significantly associated with poor overall-survival (p=0.01), and also revealed distinct enriched oncogenic pathways. However, high expression of cytotoxic gene-signature within TBX21-subgroup showed poor clinical-outcome (p=0.05). In AITL, high expression of pan B-cell signatures correlated with favorable-outcome (p=0.01), whereas high monocytic-signature was associated with inferior-survival (p=0.017). Combined prognostic score was predictive of AITL survival in independent cohort (p=0.004), suggesting role of tumor microenvironment.
    Blood 03/2014; 123(19). DOI:10.1182/blood-2013-11-536359 · 9.78 Impact Factor
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    ABSTRACT: Constitutive activation of NF-κB is a hallmark of the activated B cell-like (ABC) subtype of diffuse large B cell lymphoma (DLBCL), owing to upstream signals from the B cell receptor (BCR) and MyD88 pathways. The linear polyubiquitin chain assembly complex (LUBAC) attaches linear polyubiquitin chains to IκB kinase γ, a necessary event in some pathways that engage NF-κB. Two germ line polymorphisms affecting the LUBAC subunit RNF31 are rare among healthy individuals (~1%) but enriched in ABC DLBCL (7.8%). These polymorphisms alter RNF31 α helices that mediate binding to the LUBAC subunit RBCK1, thereby increasing RNF31-RBCK1 association, LUBAC enzymatic activity, and NF-κB engagement. In the BCR pathway, LUBAC associates with the CARD11/MALT1/BCL10 adapter complex and is required for ABC DLBCL viability. A stapled RNF31 α-helical peptide based on the ABC DLBCL-associated Q622L polymorphism inhibited RFN31-RBCK1 binding, decreased NF-κB and killed ABC DLBCL cells, credentialing this protein-protein interface as a therapeutic target.
    Cancer Discovery 02/2014; 4(4). DOI:10.1158/2159-8290.CD-13-0915 · 15.93 Impact Factor
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    ABSTRACT: The assignment of diffuse large B-cell lymphoma into cell-of-origin (COO) groups is becoming increasingly important with the emergence of novel therapies that have selective biological activity in germinal center B-cell-like (GCB) or activated B-cell-like (ABC) groups. The LLMPP's Lymph2Cx assay is a parsimonious digital gene-expression (NanoString) based test for COO assignment in formalin-fixed paraffin-embedded tissue (FFPET) routinely produced in standard diagnostic processes. The 20-gene assay was trained using 51 FFPET biopsies; the locked assay was then validated using an independent cohort of 68 FFPET biopsies. Comparisons were made with COO assignment using the original COO model on matched frozen tissue. In the validation cohort the assay was accurate, with only one case with definitive COO being incorrectly assigned, and robust, with >95% concordance of COO assignment between 2 independent laboratories. These qualities, along with the rapid turn-around-time, make Lymph2Cx attractive for implementation in clinical trials and, ultimately, patient management.
    Blood 01/2014; DOI:10.1182/blood-2013-11-536433 · 9.78 Impact Factor
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    ABSTRACT: The immune response has been implicated in the control of uveal melanoma progression. Epigenetic mechanisms mediated by specific microRNAs (miRs) regulate immune responses. Blood was drawn from six patients with uveal melanoma followed from diagnosis, at which time there was no clinical or radiographic evidence of metastasis, until metastasis manifested. Circulating T cell, natural killer (NK), natural killer T (NKT), and myeloid suppressor cell populations were assessed by flow cytometry. CD3(+), CD15(+), and CD56(+) cells were isolated using immunomagnetic beads. Plasma and cellular levels of immune regulatory miRs were determined by quantitative polymerase chain reaction assays. The development of metastasis was associated with decreases in circulating CD3(-)CD56(dim) NK cells and CD8(+) and double-negative CD3(+)CD56(+) NKT cells. ICOS(+)CD4(+)FoxP3(+) T regulatory cells and CD11b(+)CD14(-)CD15(+) myeloid suppressor cells increased. Plasma levels of miR-20a, 125b, 146a, 155, 181a, and 223 were higher in the study patients at diagnosis compared to controls. Plasma levels of miR-20a, 125b, 146a, 155, and 223 increased, and miR-181a decreased when metastasis manifested. Alterations in immune regulatory miRs were also observed in CD3(+), CD15(+), and CD56(+) cell populations. The development of metastasis in uveal melanoma is associated with changes in immune effector and regulatory cells consistent with lessening tumor immune surveillance. These changes are associated with changes in plasma and cellular levels of immune regulatory miRs. The results may help guide uveal melanoma immunotherapy and biomarker development.
    Molecular Immunology 12/2013; 58(2):182-186. DOI:10.1016/j.molimm.2013.11.018 · 3.00 Impact Factor
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    ABSTRACT: MYC amplification occurs in post-radiation and chronic lymphedema-associated secondary angiosarcoma and some primary angiosarcomas. In this study, we tested the ability of automated dual-color dual-hapten in-situ hybridization (DISH) staining to discriminate secondary angiosarcoma from radiation-associated atypical vascular lesions (AVL), and to correlate with fluorescence (FISH) for MYC amplification. Cases of secondary angiosarcoma, including 11 biopsies and 3 excisions from 11 patients, and 5 AVL biopsies from 5 patients, were examined by FISH and DISH. DISH staining was performed using the Dual Color Open Probe software on a Ventana Benchmark XT automated slide stainer. Metallic black silver (MYC) and reference CHR8 red signals were qualitatively and semi-quantitatively enumerated for tumor nuclei. Small and large clusters of silver signals were recorded as 6 or 12 signals respectively. MYC amplification was defined as MYC/CHR8 ratio >2.0. Where tissue was available for both DISH and FISH, all secondary angiosarcoma cases demonstrated MYC amplification (11/11 = 100%) by both DISH and FISH. All AVL were negative for MYC amplification by both techniques (0/5 = 0%). In the current cohort, use of DISH identified all MYC amplified cases, and distinguished secondary angiosarcoma from AVL. DISH staining may be useful in distinguishing secondary angiosarcoma from AVL in challenging cases.
    Journal of Cutaneous Pathology 12/2013; 41(3). DOI:10.1111/cup.12278 · 1.56 Impact Factor
  • Journal of thoracic oncology: official publication of the International Association for the Study of Lung Cancer 12/2013; 8(12):e109-11. DOI:10.1097/01.JTO.0000437502.82078.9b · 4.55 Impact Factor
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    ABSTRACT: We previously reported that constitutive STAT3 activation is a prominent feature of the activated B-cell subtype of diffuse large B-cell lymphomas (ABC-DLBCL). In this study, we investigated whether STAT3 activation can risk stratify patients with DLBCL. By an immunohistochemical method, we investigated phosphotyrosine STAT3 (PY-STAT3) expression from 185 patients with DLBCL treated with R-CHOP (rituximab plus cyclophosphamide, doxorubicin, vincristine, and prednisone). Cell line-based siRNA experiments were also performed to generate an 11-gene, PY-STAT3 activation signature, which was used to study a previously published cohort of 222 patients with DLBCL. The STAT3 activation status determined by these two methods and by STAT3 mRNA levels were then correlated with survival. PY-STAT3 was detected in 37% of DLBCL and enriched in ABC-DLBCL cases (P = .03). PY-STAT3 positivity significantly correlated with poor overall survival (OS; P = .01) and event-free survival (EFS; P = .006). Similar observations were made for high levels of STAT3 mRNA. In multivariable analysis, PY-STAT3 status (P = .02), International Prognostic Index (P = .02), and BCL2 expression (P = .046) were independent prognosticators of OS in this cohort. Among the cell-of-origin subgroups, PY-STAT3 was associated with poor EFS among non-germinal center B-cell DLBCL cases only (P = .027). Similarly, the 11-gene STAT3 activation signature correlated with poor survival in the entire DLBCL cohort (OS, P < .001; EFS, P < .001) as well as the ABC-DLBCL subgroup (OS, P = .029; EFS, P = .025). STAT3 activation correlated with poor survival in patients with DLBCL treated with R-CHOP, especially those with tumors of the ABC-DLBCL subtype.
    Journal of Clinical Oncology 11/2013; 31(36). DOI:10.1200/JCO.2013.52.8414 · 17.88 Impact Factor

Publication Stats

12k Citations
2,221.33 Total Impact Points


  • 2007–2014
    • Case Western Reserve University School of Medicine
      Cleveland, Ohio, United States
    • Ventana Systems, Inc.
      Harvard, Massachusetts, United States
    • Medical University of South Carolina
      • Department of Pathology and Laboratory Medicine (College of Medicine)
      Charleston, South Carolina, United States
    • Roswell Park Cancer Institute
      Buffalo, New York, United States
  • 1985–2014
    • Cleveland Clinic Laboratories
      Cleveland, Ohio, United States
  • 1979–2014
    • Cleveland Clinic
      • • Department of Molecular Pathology
      • • Department of Pathobiology
      • • Department of Urology
      Cleveland, Ohio, United States
  • 2013
    • Houston Methodist Hospital
      Houston, Texas, United States
    • University of Novi Sad
      • Faculty of Medicine
      Varadinum Petri, Autonomna Pokrajina Vojvodina, Serbia
  • 1998–2011
    • Lerner Research Institute
      Cleveland, Ohio, United States
    • University of Salzburg
      Salzburg, Salzburg, Austria
  • 2010
    • National Institutes of Health
      • Branch of Metabolism
      Bethesda, MD, United States
  • 2009
    • University of Southampton
      Southampton, England, United Kingdom
  • 2008
    • Emory University
      • Department of Pathology and Laboratory Medicine
      Atlanta, Georgia, United States
  • 2005–2008
    • Case Western Reserve University
      Cleveland, Ohio, United States
  • 2006
    • Tel Aviv University
      Tell Afif, Tel Aviv, Israel
    • St. Jude Medical
      Little Canada, Minnesota, United States
  • 2001
    • Michigan Institute of Urology
      Detroit, Michigan, United States
  • 2000
    • Albany Medical College
      Albany, New York, United States
  • 1994
    • Moffitt Cancer Center
      Tampa, Florida, United States
  • 1980
    • William Beaumont Army Medical Center
      El Paso, Texas, United States