Abstract Context.—Clotted blood and bone marrow specimens account for a large proportion of failed cytogenetic studies. There are no published protocols describing salvage of clotted specimens such that conventional chromosome or fluorescence in situ hybridization (FISH) studies can be performed. Objective.—To evaluate the utility of thrombolytic drugs on clotted blood samples to yield intact cells suitable for cytogenetic analysis. Design.—Five commercially available thrombolytic drugs (alteplase, urokinase, streptokinase, tenecteplase, reteplase) were evaluated in a series of blinded experiments to identify the best drug for lysing clots to produce samples suitable for chromosome and FISH studies. After the selection of alteplase as the drug yielding the most promising results, a comparative study between alteplase (0.75 mg/ml) and a commercially available anticlotting reagent (ACR) was performed. For each sample, mitotic index, chromosome length, and quality of slides prepared for conventional chromosome and FISH analyses were evaluated. Results.—Alteplase-treated samples produced a higher mitotic index than those treated with ACR while showing equivalent quality in conventional chromosome and FISH studies. We have demonstrated the utility of treating clotted blood samples with alteplase before cell culture to yield cells suitable for cytogenetic analysis. Since clinical implementation, this technique has been applied to more than 250 bone marrow samples, with a 93% success rate. Conclusions.—We believe the routine use of alteplase on clotted blood and bone marrow specimens should become standard for cytogenetics laboratories and may have similar utility in salvaging clotted specimens for other clinical assays requiring intact cells for analysis.
Monosomal karyotype (MK) has recently been associated with poor prognosis in acute myeloid leukemia (AML), myelodysplastic syndromes (MDS), and primary myelofibrosis (PMF). The objectives of the current study were to describe the prevalence and spectrum of autosomal monosomies in an unselected cohort of patients with known or suspected hematologic malignancies. Bone marrow cytogenetic studies (24,262) were performed at our institution between 1989 and 2009. An abnormal karyotype was demonstrated in 6,565 cases (~27%); of these, 1,365 (~21%) included autosomal monosomies that occurred as sole (n = 133; ~10%), part of two (n = 82; ~6%) or more (n = 1,150; ~84%) anomalies. All 22 autosomes were involved, but monosomy 7 was by far the most frequent, constituting ~80% of all isolated monosomies and the highest fraction of those with two or more abnormalities. Other recurrent sole monosomies included chromosomes 20 (~11%) and 21 (~4%). Monosomy 13 (~10%), 20 (~8%), 18 (~7%), 17 (~6%), 21 (~5%), 5 (~5%), and 12 (~4%) were also recurrent in the setting of ≥2 abnormalities. Bone marrow histology and clinical information were reviewed in all cases with isolated monosomy; associated clinical phenotypes were MDS (n = 60; 52 were -7), AML (n = 32; 31 were -7), myeloproliferative neoplasms (n = 16; 10 were -7), chronic myelomonocytic leukemia (CMML; n = 10; 9 were -7) and other nonmyeloid malignancies (n = 15; 4 were -7). Sole monosomy 20 (n = 14; six MDS, five MPN, and three nonmyeloid) was not seen in AML or CMML. Sole monosomy 21 was more frequent in nonmyeloid as opposed to myeloid cases.
Abstract Background: Cutaneous T-cell neoplasms display markedly heterogeneous clinical behavior despite often similar pathologic features. For example, transformed mycosis fungoides (tMF) has a poor prognosis and requires intensive therapy whereas lymphomatoid papulosis (LyP) is an indolent lymphoproliferative disorder with spontaneously resolving lesions. These entities have overlapping histologic features and no molecular tools currently exist to aid in subclassification. Methods: We identified 11 T-cell neoplasms with histologic appearances distinct from previously described cutaneous T-cell neoplasms but with features overlapping those of both tMF and LyP. The phenotypic and genetic characteristics of the tumors were studied using immunohistochemistry and fluorescence in situ hybridization, respectively. Clinical data on presentation, treatment, evolution of the lesions, and follow-up were obtained. Results: All patients were older adults (mean age, 75 yr; range, 67-88 yr; 9M:2F). Clinical presentations were similar, with papulonodular skin lesions (0.3-1.0 cm) restricted to a single body area in all but two patients. Histologically there was an extensive atypical lymphoid infiltrate in the epidermis and a dermal tumor containing large transformed cells with numerous mitotic figures and apoptotic bodies. All tumors had a T-cell phenotype and expressed CD30 in both dermal and epidermal components. No case expressed ALK. Most cases had a high proliferative rate (>80%) by Ki67 staining. FISH analysis demonstrated rearrangements of the DUSP22-IRF4 locus on 6p25.3 in all cases. No patient had evidence of systemic disease on imaging studies. In 4 untreated patients, papules began to regress spontaneously a few weeks after presentation, with complete resolution within a month or two. Seven patients were treated with local excision (with or without cryotherapy) or radiotherapy. All non-excised lesions ultimately regressed. All patients were alive at last follow-up (median, 21 mo; range, 7-150 mo). Cutaneous recurrences occurred in 5 patients. At last follow-up only 1 patient had active lesions, which were confined to the presenting site. The remaining 10 patients had been disease-free from 2 to 32 months. Conclusions: Despite pathologic features mimicking an aggressive lymphoma, the benign clinical course observed in all 11 patients suggests these cases represent a newly recognized subtype of LyP. These neoplasms are characterized by 6p25.3 rearrangements, representing the first recurrent genetic abnormality identified in LyP. In contrast, we previously have shown absence of these rearrangements in tMF. Distinction of LyP from tMF is critical in this older patient population unlikely to tolerate intensive therapy. Thus, detection of 6p25.3 rearrangements represents a new tool for molecular classification of cutaneous T-cell neoplasms. Citation Format: Andrew L. Feldman, Laszlo J. Karai, Marshall E. Kadin, Eric D. Hsi, Jason C. Sluzevich, Rhett P. Ketterling, Ryan A. Knudson. Discovery of a previously undescribed cutaneous T-cell neoplasm with chromosomal rearrangements of 6p25.3. [abstract]. In: Proceedings of the 104th Annual Meeting of the American Association for Cancer Research; 2013 Apr 6-10; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2013;73(8 Suppl):Abstract nr 1208. doi:10.1158/1538-7445.AM2013-1208
Abstract The t(2;14)(p16;q32) has been reported previously in only 12 cases of chronic lymphocytic leukemia/small lymphocytic lymphoma (CLL/SLL). The clinicopathologic features have been incompletely described. We describe 6 new cases of CLL/SLL with t(2;14)(p16;q32). All had marrow involvement, 4 had absolute lymphocytosis, 4 had lymphadenopathy, and 3 had hepatosplenomegaly. All showed atypical lymphocyte morphologic features with plasmacytoid differentiation and irregular nuclei; 3 had increased prolymphocytes. Flow cytometry demonstrated typical immunophenotypes in 5 and an atypical immunophenotype in 1. All expressed ZAP70; 5 assessed showed unmutated IgVH genes. Karyotyping identified t(2;14)(p16;q32) as the sole abnormality in 1, primary abnormality in 2, and part of a complex karyotype in 3. Fluorescence in situ hybridization analysis revealed BCL11A/IgH rearrangement in all. After chemotherapy, 3 patients died of disease and 3 were alive with disease (median follow-up, 80 months). We conclude that CLL/SLL with t(2;14) (p16;q32) and BCL11A/IgH rearrangement is characterized by atypical morphologic features and unmutated IgVH genes.
Surveys that evaluated antihypertensive drug-prescribing patterns were mailed to 150 family physicians and 150 primary care internists. The initial mailing was followed by a telephone follow-up and a second mailing. Forty-seven percent of family physicians and 41.9% of the internists who were still in practice returned the questionnaire. When asked about their choice of antihypertensive drug therapy for specific patients (based upon age, race, sex, and coexisting disease), the responses of the two specialties were similar. The only statistically significant difference was observed in the responses for a 58-year-old obese white woman with diabetes and renal impairment. In this example, the family physicians were more likely than internists to recommend an angiotensin converting enzyme inhibitor or a beta-blocker (P = .036). This study demonstrates that the majority of physicians individualized initial therapy for hypertension to the specific patient rather than strictly following a stepped-care approach with diuretics or beta-blockers as initial therapy.
