Olivier Spertini

Pulse treatment of human vascular endothelial cells with high doses of tumor necrosis factor and interferon-gamma results in simultaneous synergistic and reversible effects on proliferation and morphology.

Regional administration of high doses of tumor necrosis factor (TNF), interferon gamma (IFNγ) and melphalan to patients with advanced cancers of the limbs, results in rapid and specific tumor necrosis, while the normal adjacent tissues remain unaffected. The tumor vasculature is selectively destroyed by this treatment, and neovascular endothelial cells appear to be an early and specific target of TNF and IFNγ. To further understand some of the cellular events underlying these in vivo effects, we have investigated the response of human macro‐ and microvascular endothelial cells in vitro, after exposure to high doses of TNF and IFNγ (up to 40 × 103 U/ml each). TNF and IFNγ synergistically inhibited endothelial‐cell proliferation by up to 80% after 72 hr of treatment. Achievement of synergy required the simultaneous presence of both cytokines. A cytokine pulse as short as 30 min was sufficient to induce maximal growth inhibition measured after 48 hr. Both cytokines also induced progressive and dose‐dependent elongation of the endothelial‐cell morphology. The effects on endothelial‐cell proliferation and morphology were reversible upon removal of the cytokines. Moreover, replating of treated cells onto a fresh substrate immediately resulted in re‐acquisition of their normal shape. In contrast to the effect on cell proliferation, there was little or no effect on the rate of endothelial‐cell apoptosis. The presented data extend reports on the effects of TNF and IFNγ on human endothelial cells in vitro, and suggest that the in vivo disruption of the tumor vasculature caused by high doses of TNF and IFNγ is not due to a direct cytotoxic effect on endothelial cells but occurs through an indirect mechanism. Int. J. Cancer 77:592–599, 1998.

Plasminogen activators play an essential role in extracellular-matrix invasion by lymphoblastic T cells

Involvement of extravascular sites, in particular infiltration of the central nervous system, is a frequent complication of T‐lymphoblastic leukemia and contributes to leukemia‐associated morbidity. In this report, we studied the contribution of plasminogen activators to the invasive properties of 7 human T‐cell lines in a model of transmigration through an extracellular matrix. The T‐cell lines were found to express either urokinase (u‐PA) and high levels of u‐PA receptor or tissue‐type plasminogen activator (t‐PA) and low levels of u‐PA receptor. The rate of transmigration was consistently higher for u‐PA‐expressing cells than for t‐PA‐expressing cells. PA‐inhibitor type I (PAI‐1) was detected in the conditioned medium of one cell line and PAI‐2 was detected in cell extracts from 6 lines. The transmigration of 6 out of 7 cell lines was inhibited by trasylol, an inhibitor of plasmin, by an excess of exogenous PAI‐1 or PAI‐2, and by antibodies to the particular PA type expressed by the cells. Partial inhibition of transmigration by the amino‐terminal fragment of u‐PA implies that the u‐PA receptor contributes to transmigration. Thus, the transmigration of T‐leukemia cells through a barrier of extracellular matrix requires PA‐dependent proteolysis, which can be provided either by u‐PA or t‐PA. Specific inhibition of the PA system could provide a means to inhibit tissue invasion by T lymphoblastic cells. Int. J. Cancer, 70:461–466, 1997.

Therapeutic value of clofarabine in younger and middle-aged (18-65 years) adults with newly diagnosed AML

Clofarabine has demonstrated antileukemic activity in acute myeloid leukemia (AML) but has yet to be critically evaluated in younger adults in the frontline with standard chemotherapy. We compared 2 induction regimens in newly diagnosed patients ages 18 to 65 with acute myeloid leukemia (AML)/high-risk myelodysplastic syndromes, that is, idarubicine-cytarabine (cycle I) and amsacrine-cytarabine (cycle II) without or with clofarabine (10 mg/m2 on days 1-5 of each of both cycles). Consolidation involved chemotherapy with or without hematopoietic stem cell transplantation. Event-free survival (EFS, primary endpoint) and other clinical endpoints and toxicities were assessed. We randomized 402 and 393 evaluable patients to the control or clofarabine induction treatment arms. Complete remission rates (89%) did not differ but were attained faster with clofarabine (66% vs 75% after cycle I). Clofarabine added grades 3 to 4 toxicities and delayed hematological recovery. At a median follow-up of 36 months, the study reveals no differences in overall survival and EFS between the control (EFS, 35% ± 3 [standard error] at 4 years) and clofarabine treatments (38% ± 3) but a markedly reduced relapse rate (44% ± 3 vs 35% ± 3) in favor of clofarabine and an increased death probability in remission (15% ± 2 vs 22% ± 3). In the subgroup analyses, clofarabine improved overall survival and EFS for European Leukemia Net (ELN) 2010 intermediate I prognostic risk AML (EFS, 26% ± 4 vs 40% ± 5 at 4 years; Cox P = .002) and for the intermediate risk genotype NPM1 wild-type/FLT3 without internal-tandem duplications (EFS, 18% ± 5 vs 40% ± 7; Cox P < .001). Clofarabine improves survival in subsets of intermediate-risk AML only. HOVON-102 study is registered at Netherlands Trial Registry #NTR2187.

Chemotherapy is successful in sporadic late onset nemaline myopathy (SLONM) with monoclonal gammopathy.

Sporadic late onset nemaline myopathy (SLONM), described in 1966, is a rare proximal myopathy that is thought to be ‘‘degenerative,’’ although it is sometimes associated with human immunodeficiency virus (HIV) infection or monoclonal gammopathy. Diagnosis is based on muscle biopsy. The reported prognosis for SLOMN in gammopathy-associated forms (SLONM-MG) was found to be unfavorable despite immunotherapy, with a mortality of 80%. We report here a new SLONM-MG patient who was treated successfully with chemotherapy. A previously healthy 54-year-old man presented with a 2-year history of fatigue and progressive painless difficulty in walking and climbing stairs. He displayed proximal atrophy and weakness in scapular and pelvic girdles and in truncal muscles. He could not raise his head nor sit and stand. Needle electromyography was compatible with a myopathy in proximal muscles with no spontaneous activity at rest. Deltoid muscle biopsy showed scattered atrophic fibers. Trichome stain showed small fuschinophilic corpuscles (Fig. 1a), which were found to be nemaline rods on electron microscopy (Fig. 1b). Western blot analysis for common muscular dystrophies was normal. Work-up revealed an IgG monoclonal gammopathy (11 g/L) without signs of multiple myeloma or HIV infection. SLONM-MG was diagnosed. In the following weeks, the patient became restricted to a wheelchair and developed dyspnea. We decided to pursue aggressive therapy, as described during a meeting at the Myology Institute of Paris (2007). The patient underwent a single course of high-dose chemotherapy (melphalan 200 mg/m) and subsequent autologous peripheral blood stem-cell transplantation (PBSCT). Improvement, as documented by clinical scores (Fig. 1c), began 1 month after the graft. One year later, he could walk and stand alone, and was able to raise his arms overhead, was autonomous in daily activities, and the monoclonal protein was undetectable (Fig. 1d). This is the third SLONM-MG patient who returned to almost normal muscle function after treatment with a regimen of high-dose chemotherapy and PBSCT. Until recently, immunotherapy seemed disappointing; plasma exchange, prednisone associated with cyclophosphamide, or azathioprine, and rituximab were ineffective. The cause of SLONM is unknown, but given its frequent association with HIV infection or gammopathy, immune dysregulation has been suspected. A direct relationship between gammopathy and rod formation could not be established experimentally. Nevertheless, the present case, together with the first 2 cases, demonstrates a clear temporal correlation between presence of gammopathy and muscle impairment, probably secondary to an abnormal interaction between the circulating immunoglobulins and the sarcomeric proteins of the muscle

Dermatomyositis, lobar panniculitis and inflammatory myopathy with abundant macrophages

Dermatomyositis (DM) is a rare treatable muscle disorder with a reported favorable outcome in most patients. A localized skin/muscle involvement in a DM patient raises questions of definition and causes such as lymphoma, or relapse. We describe here a young treated DM patient who presented a focal biopsy-proven destructive myositis and dermatitis restricted to the left thigh 15 months after the onset of a treated dermatomyositis. There was evidence of subcutaneous lobular panniculitis, somewhat resembling cytophagic histocytic panniculitis associated with a focal inflammatory myopathy with abundant macrophages that destroyed the sartorius muscle. Mild signs of hemophagocytosis and T-CD3 lymphocytosis were present in the bone marrow, but no monoclonal T-lymphocyte expansion was observed, as searched by autoradiography of the totality of TcR Vgamma families. The patient improved with prednisone and azathioprine. We conclude that this focal complication suggests a continuum between dermatomyositis, CHP, and IMAM, the three syndromes where T-cell activation plays an important role.

Transfusion independence and survival in patients with acute myeloid leukemia treated with 5-azacytidine

Reanalysis of the CALGB[1][1] and AZA001[2][2] studies in advanced myelodysplastic syndrome (MDS) suggests that 5-azacytidine (AZA) is effective for acute myeloblastic leukemia (AML) with less than 30% bone marrow blasts. Most AML patients are elderly (>65 years old) and unfit for intensive

Using Digital RNA Counting and Flow Cytometry to Compare mRNA with Protein Expression in Acute Leukemias

Background The diagnosis of malignant hematologic diseases has become increasingly complex during the last decade. It is based on the interpretation of results from different laboratory analyses, which range from microscopy to gene expression profiling. Recently, a method for the analysis of RNA phenotypes has been developed, the nCounter technology (Nanostring® Technologies), which allows for simultaneous quantification of hundreds of RNA molecules in biological samples. We evaluated this technique in a Swiss multi-center study on eighty-six samples from acute leukemia patients. Methods mRNA and protein profiles were established for normal peripheral blood and bone marrow samples. Signal intensities of the various tested antigens with surface expression were similar to those found in previously performed Affymetrix microarray analyses. Acute leukemia samples were analyzed for a set of twenty-two validated antigens and the Pearson Correlation Coefficient for nCounter and flow cytometry results was calculated. Results Highly significant values between 0.40 and 0.97 were found for the twenty-two antigens tested. A second correlation analysis performed on a per sample basis resulted in concordant results between flow cytometry and nCounter in 44–100% of the antigens tested (mean = 76%), depending on the number of blasts present in a sample, the homogeneity of the blast population, and the type of leukemia (AML or ALL). Conclusions The nCounter technology allows for fast and easy depiction of a mRNA profile from hematologic samples. This technology has the potential to become a valuable tool for the diagnosis of acute leukemias, in addition to multi-color flow cytometry.

Eosinophilic perimyositis as the presenting feature of a monoclonal T-cell expansion

A 51‐year‐old physically active man was investigated for exertional myalgias and muscle stiffness. On examination he had mild proximal muscle weakness of the upper extremities and retraction of the digit flexors. Blood eosinophilia was present, but serum creatine kinase (CK) levels and an electromyographic study were normal. A skin–fascia–muscle biopsy of the calf revealed a macrophagic and CD4+T‐cell infiltration of the perimysium, and a T‐cell expansion was observed in blood, bone marrow, and muscle. A diagnosis of eosinophilic perimyositis was made, and prednisone and azathioprine were administrated with a good clinical response. This case highlights the differential diagnosis of blood eosinophilia with muscle disorders, and underscores that eosinophilic perimyositis may be the expression of a T‐cell monoclonal expansion. Although the pathogenesis behind the T‐cell expansion is unclear but probably inflammatory, we suggest regular follow‐up to allow early treatment of any T‐cell lymphoproliferative malignancy that may develop. Muscle Nerve, 2005

Evolving characteristics and outcome of secondary acute promyelocytic leukemia (APL): A prospective analysis by the French‐Belgian‐Swiss APL group

Reports of patients with secondary acute promyelocytic leukemia (APL) have increased in recent years, particularly for those who received treatment with mitoxantrone, and retrospective studies have suggested that their characteristics and outcomes were similar to those of patients with de novo APL.

Pentraxin-3 polymorphisms and invasive mold infections in acute leukemia patients with intensive chemotherapy

Invasive mold infections are a major concern in oncohematologic patients, with incidence and mortality rates ranging between 15–30%.[1][1] While mold-active prophylaxis is recommended during induction chemotherapy for acute leukemia, its universal use has been challenged based on the large number