Giuseppe Montalbano

Deletion of chromosome 20 in bone marrow of patients with Shwachman‐Diamond syndrome, loss of the EIF6 gene and benign prognosis

Bianchi, V., Robles, R., Alberio, L., Furlan, M. & Lammle, B. (2002) Von Willebrand factor-cleaving protease (ADAMTS13) in thrombocytopenic disorders: a severely deficient activity is specific for thrombotic thrombocytopenic purpura. Blood, 100, 710–713. Coppo, P., Bengoufa, D., Veyradier, A., Wolf, M., Bussel, A., Millot, G.A., Malot, S., Heshmati, F., Mira, J.P., Boulanger, E., Galicier, L., DureyDragon, M.A., Fremeaux-Bacchi, V., Ramakers, M., Pruna, A., Bordessoule, D., Gouilleux, V., Scrobohaci, M.L., Vernant, J.P., Moreau, D., Azoulay, E., Schlemmer, B., Guillevin, L. & Lassoued, K. (2004) Severe ADAMTS13 deficiency in adult idiopathic thrombotic microangiopathies defines a subset of patients characterized by various autoimmune manifestations, lower platelet count, and mild renal involvement. Medicine (Baltimore), 83, 233–244. Froehlich-Zahnd, R., George, J.N., Vesely, S.K., Terrell, D.R., Aboulfatova, K., Dong, J.F., Luken, B.M., Voorberg, J., Budde, U., Sulzer, I., Lammle, B. & Kremer Hovinga, J.A. (2011) Evidence for a role of anti-ADAMTS13 autoantibodies despite normal ADAMTS13 activity in recurrent thrombotic thrombocytopenic purpura. Haematologica. Epub ahead of print. doi: 10.3324/haematol.2011.051433. Hovinga, J.A., Vesely, S.K., Terrell, D.R., Lammle, B. & George, J.N. (2010) Survival and relapse in patients with thrombotic thrombocytopenic purpura. Blood, 115, 1500–1511; quiz 1662. Legendre, C.M., Babu, S., Furman, R.R., Sheerin, N.S., Cohen, D.J., Gaber, O., Eitner, F., Delmas, Y., Loirat, C., Greenbaum, L.A. & Zimmerhackl, L.B. (2010) Safety & efficacy of eculizumab in aHUS patients resistant to plasma therapy: interim analysis from a phase II trial. Journal of the American Society of Nephrology, 21, 93A. Mache, C.J., Acham-Roschitz, B., Fremeaux-Bacchi, V., Kirschfink, M., Zipfel, P.F., Roedl, S., Vester, U. & Ring, E. (2009) Complement inhibitor eculizumab in atypical hemolytic uremic syndrome. Clin J Am Soc Nephrol, 4, 1312– 1316. Remuzzi, G. (2003) Is ADAMTS-13 deficiency specific for thrombotic thrombocytopenic purpura? No. J Thromb Haemost, 1, 632– 634. Remuzzi, G., Galbusera, M., Noris, M., Canciani, M.T., Daina, E., Bresin, E., Contaretti, S., Caprioli, J., Gamba, S., Ruggenenti, P., Perico, N. & Mannucci, P.M. (2002) von Willebrand factor cleaving protease (ADAMTS13) is deficient in recurrent and familial thrombotic thrombocytopenic purpura and hemolytic uremic syndrome. Blood, 100, 778–785. Salmon, J.E., Heuser, C., Triebwasser, M., Liszewski, M.K., Kavanagh, D., Roumenina, L., Branch, D.W., Goodship, T., Fremeaux-Bacchi, V. & Atkinson, J.P. (2011) Mutations in complement regulatory proteins predispose to preeclampsia: a genetic analysis of the PROMISSE cohort. PLoS Med, 8, e1001013. Tsai, H.M. (2003) Is severe deficiency of ADAMTS-13 specific for thrombotic thrombocytopenic purpura? Yes. Journal of Thrombosis and Haemostasis, 1, 625–631.

Different loss of material in recurrent chromosome 20 interstitial deletions in Shwachman-Diamond syndrome and in myeloid neoplasms

BackgroundAn interstitial deletion of the long arms of chromosome 20, del(20)(q), is frequent in the bone marrow (BM) of patients with myelodysplastic syndromes (MDS), acute myeloid leukemia (AML), and myeloproliferative neoplasms (MPN), and it is recurrent in the BM of patients with Shwachman-Diamond syndrome (SDS), who have a 30-40% risk of developing MDS and AML.ResultsWe report the results obtained by microarray-based comparative genomic hybridization (a-CGH) in six patients with SDS, and we compare the loss of chromosome 20 material with one patient with MDS, and with data on 92 informative patients with MDS/AML/MPN and del(20)(q) collected from the literature.ConclusionsThe chromosome material lost in MDS/AML/MPN is highly variable with no identifiable common deleted regions, whereas in SDS the loss is more uniform: in 3/6 patients it was almost identical, and the breakpoints that we defined are probably common to most patients from the literature. In some SDS patients less material may be lost, due to different distal breakpoints, but the proximal breakpoint is in the same region, always leading to the loss of the EIF6 gene, an event which was related to a lower risk of MDS/AML in comparison with other patients.

High variability of genomic instability and gene expression profiling in different HeLa clones.

The HeLa cell line is one of the most popular cell lines in biomedical research, despite its well-known chromosomal instability. We compared the genomic and transcriptomic profiles of 4 different HeLa batches and showed that the gain and loss of genomic material varies widely between batches, drastically affecting basal gene expression. Moreover, different pathways were activated in response to a hypoxic stimulus. Our study emphasizes the large genomic and transcriptomic variability among different batches, to the point that the same experiment performed with different batches can lead to distinct conclusions and irreproducible results. The HeLa cell line is thought to be a unique cell line but it is clear that substantial differences between the primary tumour and the human genome exist and that an indeterminate number of HeLa cell lines may exist, each with a unique genomic profile.

Cytogenetic monitoring in Shwachman-Diamond syndrome: a note on clonal progression and a practical warning.

We analyzed the results of periodic chromosome analyses performed on bone marrow of 22 patients with Shwachman-Diamond syndrome (SDS), 8 directly observed and 14 from the literature, selected because of changes in the cytogenetic picture during the course of the disease. This study points out some features of the cytogenetic evolution in SDS relevant for prognostic evaluation but never noted in the literature. In particular, the lack of any clonal progression and the frequent appearance of independent clones with chromosomal changes different from the one initially discovered, with possible severe prognostic implications, are reported.

Improving the definition of the structure of the isochromosome i(7)(q10) in Shwachman-Diamond Syndrome

Harada, M., Sumida, I., Hanada, M. & Niho, Y. (1990) Central nervous system involvement in adult T-cell leukemia/lymphoma. Cancer, 65, 327–332. Umehara, F., Izumo, S., Ronquillo, A.T., Matsumuro, K., Sato, E. & Osame, M. (1994) Cytokine expression in the spinal cord lesions in HTLV-I-associated myelopathy. Journal of Neuropathology and Experimental Neurology, 53, 72–77. Wu, J.M., Georgy, M.F., Burroughs, F.H., Weir, E.G., Rosenthal, D.L. & Ali, S.Z. (2009) Lymphoma, leukemia, and pleiocytosis in cerebrospinal fluid: is accurate cytopathologic diagnosis possible based on morphology alone? Diagnostic Cytopathology, 37, 820– 824.

Chromosome anomalies in bone marrow as primary cause of aplastic or hypoplastic conditions and peripheral cytopenia: disorders due to secondary impairment of RUNX1 and MPL genes

BackgroundChromosome changes in the bone marrow (BM) of patients with persistent cytopenia are often considered diagnostic for a myelodysplastic syndrome (MDS). Comprehensive cytogenetic evaluations may give evidence of the real pathogenetic role of these changes in cases with cytopenia without morphological signs of MDS.ResultsChromosome anomalies were found in the BM of three patients, without any morphological evidence of MDS: 1) an acquired complex rearrangement of chromosome 21 in a boy with severe aplastic anaemia (SAA); the rearrangement caused the loss of exons 2–8 of the RUNX1 gene with subsequent hypoexpression. 2) a constitutional complex rearrangement of chromosome 21 in a girl with congenital thrombocytopenia; the rearrangement led to RUNX1 disruption and hypoexpression. 3) an acquired paracentric inversion of chromosome 1, in which two regions at the breakpoints were shown to be lost, in a boy with aplastic anaemia; the MPL gene, localized in chromosome 1 short arms was not mutated neither disrupted, but its expression was severely reduced: we postulate that the aplastic anaemia was due to position effects acting both in cis and in trans, and causing Congenital Amegakaryocytic Thrombocytopenia (CAMT).ConclusionsA clonal anomaly in BM does not imply per se a diagnosis of MDS: a subgroup of BM hypoplastic disorders is directly due to chromosome structural anomalies with effects on specific genes, as was the case of RUNX1 and MPL in the patients here reported with diagnosis of SAA, thrombocytopenia, and CAMT. The anomaly may be either acquired or constitutional, and it may act by deletion/disruption of the gene, or by position effects. Full cytogenetic investigations, including a-CGH, should always be part of the diagnostic evaluation of patients with BM aplasia/hypoplasia and peripheral cytopenias.

Bone marrow failure may be caused by chromosome anomalies exerting effects on RUNX1T1 gene

BackgroundThe majority of the cases of bone marrow failure syndromes/aplastic anaemias (BMFS/AA) are non-hereditary and considered idiopathic (80–85%). The peripheral blood picture is variable, with anaemia, neutropenia and/or thrombocytopenia, and the patients with idiopathic BMFS/AA may have a risk of transformation into a myelodysplastic syndrome (MDS) and/or an acute myeloid leukaemia (AML), as ascertained for all inherited BMFS. We already reported four patients with different forms of BMFS/AA with chromosome anomalies as primary etiologic event: the chromosome changes exerted an effect on specific genes, namely RUNX1, MPL, and FLI1, leading to the disease.ResultsWe report two further patients with non-hereditary BM failure, with diagnosis of severe aplastic anaemia and pancytopenia caused by two different constitutional structural anomalies involving chromosome 8, and possibly leading to the disorder due to effects on the RUNX1T1 gene, which was hypo-expressed and hyper-expressed, respectively, in the two patients. The chromosome change was unbalanced in one patient, and balanced in the other one.ConclusionsWe analyzed the sequence of events in the pathogenesis of the disease in the two patients, including a number of non-haematological signs present in the one with the unbalanced anomaly. We demonstrated that in these two patients the primary event causing BMFS/AA was the constitutional chromosome anomaly. If we take into account the cohort of 219 patients with a similar diagnosis in whom we made cytogenetic studies in the years 2003–2017, we conclude that cytogenetic investigations were instrumental to reach a diagnosis in 52 of them. We postulate that a chromosome change is the primary cause of BMFS/AA in a not negligible proportion of cases, as it was ascertained in 6 of these patients.