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E Dmitrovsky

Publications and source records attributed to E Dmitrovsky.

At least 91 records · Page 5Linked to original sources

Oncogenes and clinical oncology.

The past year has seen important advances in our understanding of the molecular biology of human cancer. We have learned more about how normal genes with critical functions in growth and development can induce cellular transformation and malignancy if mutated or overexpressed. The finding of such oncogenes in specific human cancers often portends a poor prognosis. We have learned more about tumor suppressor genes, whose loss by mutation, deletion, or translocation can lead to cancer. A series of defects involving both oncogenes and tumor suppressor genes has been shown to characterize the multistep development of a fully malignant colon cancer. We have new insights into the promotion of malignancy by the fused gene product resulting from the chromosomal abnormality diagnostic of one leukemia, chronic myelogenous leukemia. Recently, in acute promyelocytic leukemia, a characteristic chromosomal abnormality has been shown to result in a specific fusion of a nuclear receptor that activates transcription and a previously unknown gene. Most interestingly, a ligand for this rearranged receptor has been shown to be a novel effective treatment for the disease. This review summarizes many of these advances.

Animals↗

Genetic analysis of germ cell tumors: current progress and future prospects.

Cytogenetic analysis of germ cell tumors (GCTs) has identified i(12p) as a specific cytogenetic abnormality identified in more than 80% of GCTs, present in all histologies, in primary and metastatic lesions, in testicular and extragonadal presentations, and in ovarian and sex cord stromal tumors. Other nonrandom numeric and structural chromosomal abnormalities have also been identified. Oncogene studies suggest a potential role for n-ras mutations in GCT transformation. The role of loss of tumor suppressor genes and increased genomic dosage of growth promoter genes remain areas of great interest. Leukemias and differentiated malignancies that arise in the setting of GCT appear to be clonally derived from GCT cells, with evidence of karyotypic progression and acquisition of other tumor-specific cytogenetic markers. Identification of i(12p) in poorly differentiated midline carcinomas of uncertain histogenesis can assist in the diagnosis of GCT. The presence of three or more copies of 12p may predict resistance to chemotherapy and portend a higher likelihood of treatment failure. Future cytogenetic studies in GCT promise to provide insight into the biology and treatment of all solid tumors, because GCTs are a model of chemotherapeutic responsiveness, cellular differentiation, and tumor clonal evolution.

Cell Differentiation↗

The role of ifosfamide plus cisplatin-based chemotherapy as salvage therapy for patients with refractory germ cell tumors.

A prospective study of four cycles of etoposide with ifosfamide and cisplatin (VIP) chemotherapy was conducted in 42 germ cell tumor (GCT) patients who were refractory to cisplatin with etoposide/vinblastine-based therapy. Forty patients were evaluable for response. Ten patients (25%) had a complete response: seven to chemotherapy alone and an additional three patients after surgical resection of viable GCT. With a median follow-up of 15 months, four complete responders relapsed, and six patients (15%) remain in remission. Hematologic and nephrotoxicity were moderately severe. Durable complete responses with VIP as second salvage were achieved and suggests that ifosfamide adds efficacy to standard first-salvage therapy. The observed nephrotoxicity and myelotoxicity are considerations in the design of ifosfamide-cisplatin-based regimens. Hematopoietic growth factors may be useful in ameliorating myelotoxicity. The early use of ifosfamide-based chemotherapy may reduce the nephrotoxicity exacerbated by prior cisplatin. A trial of VIP as first salvage after a relapse from a complete response to platinum-based induction therapy is warranted. The modest proportion of patients who achieve a durable remission to VIP as second salvage emphasizes the need for more efficacious salvage therapy for patients who do not achieve a durable complete response.

Adolescent↗

Nonrandom chromosome abnormalities in testicular and ovarian germ cell tumor cell lines.

We report the karyotypic analysis of seven testicular and one ovarian germ cell tumor (GCT) cell lines, a number of which have previously been partially investigated. An i(12p) was found in each of the testicular GCT cell lines, while it was absent in the ovarian GCT cell line. Thus, our study extends to cell lines the observation from fresh tumor tissues that i(12p) is a highly nonrandom chromosomal abnormality of testicular GCT. Additional consistent nonrandom abnormalities in the testicular GCT cell lines included the following: del(1)(p22), del(1)(q21), i(1q), del(7)(q11.2), and del(12)(q14). The del(12)(q14) abnormality was identified in five of the cell lines investigated. This observation, together with previous detection of this marker chromosome in fresh tumor specimens by us and others, suggests that loss of genetic material on 12q may represent a primary change associated with malignant transformation of testicular germ cells. As reported in a previous study, a t(15;20)(p11;q11) translocation was identified in the ovarian GCT cell line. Interestingly, it also was seen in one testicular GCT cell line. In addition, a der(15)t(15;20)(p11;q11) marker chromosome was identified in two other testicular GCT cell lines. Thus, this reinvestigation of GCT cell lines has resolved the discrepancy regarding the occurrence of i(12p) in fresh tumors versus established cell lines and identified additional nonrandom abnormalities of potential importance to the development of GCTs.

Cell Line↗

Cytogenetic and molecular analysis of human male germ cell tumors: chromosome 12 abnormalities and gene amplification.

We report karyotypic analysis of 24 male germ cell tumors (GCTs) with clonally abnormal karyotypes biopsied from testicular and extragonadal lesions from 20 patients belonging to the histologic categories seminoma, teratoma, embryonal carcinoma, choriocarcinoma, and endodermal sinus tumor. Chromosomes 1, 7, 9, 12, 17, 21, 22, and the X chromosome were nonrandomly gained in these tumors. Nonrandom structural changes affected most frequently chromosomes 1 and 12, the latter as i(12p) and/or del(12)(q13----q22). The i(12p) was seen in 90% of tumors which included all histologic subtypes and gonadal as well as extragonadal presentation. Our present results, along with those from published data on fresh GCT biopsies, establish that i(12p) is a highly nonrandom chromosome marker of all histologic as well as anatomic presentations of GCTs. in contrast, we found del(12)(q13----q22) exclusively in nonseminomatous GCTs (NSGCTs) and mixed GCTs (MGCTs) occurring in 44% of such lesions. Because successful cytogenetic analysis of fresh tumor specimens is not always possible, we developed a method based on DNA analysis to detect i(12p) as increased copy number of 12p. In addition to the changes affecting chromosome 12 identified above, we have detected, for the first time, cytological evidence of gene amplification in the form of homogeneously staining regions (HSRs) and double minute chromosomes (dmins) in treated as well as untreated primary extragonadal and metastatic GCTs and confirmed the presence of amplified DNA in one of these tumors at the molecular level by the in-gel renaturation method. Hybridization of DNA from cultured cells from an HSR-bearing tumor with a panel of probes for genes known to be amplified or otherwise perturbed in diverse tumor systems did not identify the amplified gene, suggesting amplification of a novel gene or genes. This study comprises the largest series of GCT cytogenetics attempted so far. Notably, it includes data on a series of primary mediastinal tumors, a group which previously has not been studied in any detail.

Blotting, Southern↗

A phase II trial of carboplatin (CBDCA) in small-cell and non-small-cell lung cancer with correlation to in vitro analysis of cytotoxicity.

A Phase II trial of carboplatin (CBDCA) was performed in 33 patients with advanced lung cancer, including 15 patients with inoperable Stage III non-small-cell (NSCLC) and 18 patients with relapsed small-cell (SCLC) lung cancer. Initial dosage was 320 mg/m2 infused over 24 h; in the absence of hematologic toxicity, subsequent doses were escalated to 400 mg/m2. Patients received a median of two cycles (range 1-13 for NSCLC and 1-5 for SCLC) of therapy. There were no complete or partial responses among the NSCLC patients. Among the SCLC patients, two had a partial response. In vitro analysis of the cytotoxicity of CBDCA and its parent compound cisplatin by two different methods for 20 NSCLC cell lines suggested that equivalent tumor cell kill is achieved by the two compounds, but this occurs at a log lower concentration of cisplatin than of CBDCA. The in vitro cytotoxicity against NSCLC of CBDCA at a concentration predicted to be in the range produced by the dose employed in this Phase II study correlated well with the resulting very modest in vivo benefit. In vitro, a continuous dose-response relationship exists for CBDCA, suggesting that if higher doses could be administered safely to patients, greater clinical benefit might occur. We conclude that single agent CBDCA in the dosage and schedule administered has less than 20% activity (95% confidence intervals 0-19%) in NSCLC and an 11% response rate in SCLC (95% confidence intervals 2-34%). Despite this outcome, in vitro data in human NSCLC cell lines suggest higher dosages should perhaps be evaluated before discounting a role for CBDCA in the management of NSCLC.

Adult↗

Clinical and genetic features of human germ cell cancer.

The majority of patients with disseminated germ cell cancer can be cured. However, subsets of these patients are either refractory to initial treatment or relapse after treatment. The management of these different clinical groups is reviewed. An enhanced understanding of the basic biological features of germ cell cancer may well lead to improvements in the management of this disease. Recent advances in the biological and genetic aspects of germ cell cancer which are pertinent to the behaviour of this malignancy are summarized.

Cell Transformation, Neoplastic↗

Retinoic acid induces down-regulation of several growth factors and proto-oncogenes in a human embryonal cancer cell line.

The human teratocarcinoma cell NTERA-2 cl. D1 (NT2/D1) is a cloned embryonal cancer cell line that differentiates into a neuronal phenotype and other cellular lineages after treatment with retinoic acid (RA). We examined the regulated expression of growth factors and proto-oncogenes in NT2/D1 cells. We studied RNA levels after six days of RA treatment to assess gene expression coincident with observed morphologic differentiation. Three growth factors were markedly down-regulated following RA treatment: Hst-1/kFGF and TGF-alpha expression became undetectable by Northern analysis and bFGF expression was substantially reduced. Minimal decline was seen for c-myc, N-myc, c-fos, and c-myb. Increased expression with differentiation was seen for the human homeotic genes Hox 2.1 and Hox 2.2. Assay of RNA levels daily after one to six days of RA treatment showed that the growth factor down-regulation inversely correlated with the homeotic gene up-regulation. Nuclear run-on studies showed low transcriptional rates for these homeotic genes, Hst-1/kFGF, and TGF-alpha that did not measurably change with RA treatment. To explore whether these regulated genes in NT2/D1 play a role in human testicular cancer, we examined RNA levels in a panel of human germ cell cancer lines. Hst-1/kFGF and bFGF are commonly expressed in five of seven male germ cell cancer lines. These data show that specific proto-oncogenes and growth factors are down-regulated with RA treatment of the NT2/D1 cell and that some of these regulated genes are often expressed in human germ cell cancer lines.

Blotting, Northern↗

Isochromosome 12p in non-seminoma cell lines: karyologic amplification of c-ki-ras2 without point-mutational activation.

We examined eight human germ cell cancer lines (GCCLs) for cytogenetic abnormalities and found an isochromosome 12p, i(12p), marker in all seven male nonseminoma GCCLs, but not in the single female teratocarcinoma cell line. Southern blot analysis of these cell lines showed increased copy number for c-ki-ras2, a gene located on 12p, in all the male GCCLs. The comparison of Southern blot analysis for a restriction fragment length polymorphism (RFLP) probe localized to 12p to a probe for int-1, which maps to 12q, indicates that the increased copy number for c-ki-ras2 is primarily from the greater numbers of 12p relative to 12q. Although Northern analysis revealed enhanced mRNA expression for c-ki-ras2 in the GCCLs with an i(12p), hybridization of specific end-labelled oligonucleotides to the polymerase chain reaction products of c-ki-ras2 codons 12, 13, or 61 did not identify c-ki-ras2 mutations of these codons in these cells. Thus, c-ki-ras2 activation through point mutation is an infrequent event in GCCLs. These data further suggest that increased 12p copy number is a common event in the transformation process leading to male germ cell cancer. We conclude that determination of 12p copy number by cytogenetic analysis or Southern blotting is useful in the diagnostic evaluation of human germ cell cancer.

Blotting, Southern↗

Clinical use of differentiating agents in cancer therapy.

Conceptually, the use of cytodifferentiation agents to suppress oncogenicity may prove most effective in long-term continuous treatment of patients whose tumors display some inherent expression of differentiated characteristics. This is the case in many pre-malignant lesions. The difficulty in evaluating the therapeutic role for HMBA, the retinoids, and cytosine arabinoside is oftentimes in separating in vivo cytotoxic from cytodifferentiation effects. Only with further studies will it be determined whether these and other cytodifferentiation agents are therapeutically useful and whether they can provide additional understanding of the processes of cellular transformation and differentiation with which they interact.

Acetamides↗

Retinoic acid causes a decline in TGF-alpha expression, cloning efficiency, and tumorigenicity in a human embryonal cancer cell line.

The human teratocarcinoma NTERA-2 cl. D1 (NT2/D1) cell is a cloned multipotential embryonal cancer cell line that differentiates into a neuronal phenotype and other cellular lineages with retinoic acid (RA) treatment. Here we report that mRNA for the transforming growth factor-alpha is expressed in these RA-untreated cells and that RA-treatment results in a reduction of mRNA expression within 24 hr of treatment. In total cellular RNA, TGF-alpha mRNA is not detectable by Northern analysis at 6 days when there is increased expression of the human homeotic genes Hu-1 (Hox 2.1) and Hu-2 (Hox 2.2), known markers of RA response in NT2/D1 cells. RA treatment also causes a marked reduction in cloning efficiency and tumorigenicity of these cells. The addition of TGF-alpha or EGF (epidermal growth factor) protein to RA-untreated NT2/D1 cells augments soft agar cloning under limited fetal calf serum conditions. Blocking monoclonal antibodies directed against the EGF receptor (EGFr) can prevent this augmentation. We conclude that TGF-alpha expression inversely correlates with the state of RA-induced differentiation of this human teratocarcinoma cell and that TGF-alpha and EGF proteins are stimulatory growth factors in NT2/D1 cells under these culture conditions.

Cell Differentiation↗

Isochromosome of the short arm of chromosome 12: clinically useful markers for male germ cell tumors.

Twenty-nine tumor specimens were obtained from 24 males with germ cell tumors. All primary sites and histologies were represented. An isochromosome of the short arm of chromosome 12 [i (12p)] was found in 20 specimens obtained from 16 patients, a 46,XY normal karyotype was present in seven specimens, and one specimen was a cytogenetic failure. The i(12p) was found in tumors from all primary sites and in all histologies, including a choriocarcinoma. The presence of three or more additional copies of 12p was associated with a statistically significant greater likelihood of treatment failure. With diagnostic and possibly prognostic importance in germ cell tumors in males, the high frequency of i(12p) in our series of studies and in those of others indicates that it probably occurs as a very early defect in the development of these tumors. Further studies of chromosome 12 in males with these tumors are warranted.

Chromosome Aberrations↗

Application of RNA-RNA tissue in situ hybridization in an analysis of a patient with leukemia.

RNA-RNA tissue in situ hybridization is a relatively new technique that detects gene expression in individual cells. In this report we compare and contrast the technique with conventional biologic analysis. We illustrate how this technique could function as a diagnostic tool by applying it to a 58-year-old man with a four-month history of lymphadenopathy and peripheral lymphocytosis. RNA-RNA tissue in situ hybridization performed on sections of one of this patient's lymph nodes and on cytospins of his peripheral blood demonstrated the presence of an apparent monoclonal population of B cells producing mu and lambda immunoglobulin (Ig) messages in the lymph node and peripheral blood as well as a T-cell population in the lymph node only. These results were corroborative and complementary to conventional DNA (Southern) and RNA (Northern) analyses. The data were consistent with the diagnosis of chronic lymphocytic leukemia (CLL). With the use of this technique, an intriguing pattern of cellular heterogeneity was observed within the mu-lambda population of cells in the lymph node. A subset of these cells appeared to express a much greater amount of immunoglobulin message and to cluster around the lymph node vessels. The combination of RNA-RNA in situ hybridization and routine histopathology has the potential for providing an additional dimension to tumor analysis.

DNA, Neoplasm↗

Cytologic transformation in cutaneous T cell lymphoma: a clinicopathologic entity associated with poor prognosis.

The clinical course of cutaneous T cell lymphoma (mycosis fungoides and Sezary syndrome) is generally indolent, but in occasional patients becomes fulminant. We found that biopsies from patients with accelerating disease can reveal cytologic transformation from previously observed small, convoluted lymphocytes to large cells that are similar to cells seen in large-cell lymphoma. The cerebriform nuclei characteristic of malignant T cells can only rarely be identified. Of 150 cutaneous T cell lymphoma patients we treated from 1976 to 1984, cytologic transformation was identified in 12 after review of peripheral blood smears and biopsies from skin, lymph nodes, and visceral sites. Patients who developed cytologic transformation were initially characterized by advanced stage (11 of 12), with lymph node effacement (seven of 11) and erythroderma (five of 12). The tumor cell DNA content after transformation was aneuploid (four of four), and the ability to form rosettes with sheep erythrocytes was retained in transformed cells (three of three). The median time from diagnosis of cutaneous T cell lymphoma to cytologic transformation was 21.5 months (range, 4 to 64), and the median survival from transformation was only 2 months (range, 0 to 19+). We conclude that cytologic transformation in cutaneous T cell lymphoma represents a distinct clinicopathologic entity, characterized by an aggressive clinical course.

Biopsy↗

Identification of three new Ig lambda-like genes in man.

Three new human lambda L chain-like Ig genes are identified by restriction enzyme and nucleotide sequence analysis. Two genes, 14.1 and 16.1, have intact J and C regions, and are potentially functional, with open reading frames. A third gene, 18.1, is a pseudogene. The evolutionary lineage of these genes compared to the known functional locus lambda C1-lambda C6 can be surmised from Southern blot and nucleotide homologies. This study demonstrates that the human lambda gene family is more complex than previously recognized.

Amino Acid Sequence↗