Search

A 36‐year‐old patient with a disorder characterized by severe mental retardation, behavioral problems, dysmorphic face, “muscular build,” and hand/foot anomalies, is reported. Following a diagnosis of de novo pericentric inversion of chromosome 8 based on standard cytogenetic analysis, a subsequent 75 kb array‐CGH investigation disclosed a deletion spanning for about 13.7 Mb in the 2q31.2q32.3 region. Whole painting of chromosome 8 established the intrachromosomal nature of the rearrangement and FISH analysis with locus‐specific probes confirmed the deletion on the long arm of chromosome 2. The deleted region, clinical outcome, and medical history in this patient are mainly superimposable to those reported in a published 8‐year‐old boy, suggesting that this genomic segment is prone to rearrangements and its hemizygosity gives rise to a clinically recognizable syndrome. The role of some genes mapping in the deleted region and related with distinct disorders is discussed. Interestingly, deletion of MSTNgene, a negative regulator of muscle growth, was associated in our patient with a “muscular build,” a feature which could be regarded as a handle for clinical recognition of this syndrome. © 2009 Wiley‐Liss, Inc.

Purpose: We evaluated the experiences of 19 Italian laboratories concerning 241 small supernumerary marker chromosomes (sSMCs) with the aim of answering questions arising from their origin from any chromosome, their variable size and genetic content, and their impact on the carrier's phenotype.Methods: Conventional protocols were used to set up the cultures and chromosome preparations. Both commercial and homemade probes were used for the fluorescent in situ hybridization analyses.Results: A total of 113 of the 241 sSMCs were detected antenatally, and 128 were detected postnatally. There were 52 inherited and 172 de novo cases. Abnormal phenotype was present in 137 cases (57%), 38 of which were antenatally diagnosed. A mosaic condition was observed in 87 cases (36%). In terms of morphology, monocentric and dicentric bisatellited marker chromosomes were the most common, followed by monocentric rings and short-arm isochromosomes. The chromosomes generating the sSMCs were acrocentric in 132 cases (69%) and non-acrocentric chromosomes in 60 cases (31%); a neocentromere was hypothesized in three cases involving chromosomes 6, 8, and 15.Conclusion: The presented and published data still do not allow any definite conclusions to be drawn concerning karyotype–phenotype correlations. Only concerted efforts to characterize molecularly the sSMCs associated or not with a clinical phenotype can yield results suitable for addressing karyotype–phenotype correlations in support of genetic counseling.

We evaluated the experiences of 19 Italian laboratories concerning 241 small supernumerary marker chromosomes (sSMCs) with the aim of answering questions arising from their origin from any chromosome, their variable size and genetic content, and their impact on the carrier's phenotype.

Therapy of solid and hematologic tumors with alkylating agents appears to increase the frequency of acute non-lymphocytic leukemia (ANLL), as indicated by the cases reported in the literature. The carcinogenetic mechanism of alkylating agents seems related to their ability to damage DNA, and this is supported by the findings of multiple cytogenetic abnormalities in these patients. We report a case of ANLL secondary to therapy with melphalan, which was utilized on an adjuvant basis for breast cancer. ANLL developed 24 months after chemotherapy was discontinued. Results of the cytogenetic analysis in our patient showed multiple rearrangements and marker chromosomes. Among these was a large metacentric chromosome, identified in 6 of 8 karyotypes, in the size range of group A, which probably resulted from a translocation t(7;14) (7qter→7p11::14p11-→14qter). The natural history of the underlying disease and of the ANLL in our patient and data from chromosomal analysis seem to confirm the hypothesis that alkylating agents are potentially leukemogenic in man, probably through genetic damage. This possibility should be considered when such cytotoxic drugs are used in an adjuvant setting.

Glycosaminoglycans (GAGs), normal components of the extracellular matrix (ECM), and the glycosidases, that degrade them, play a key role in the bone remodelling process. The effects of interleukin-1α (IL-1α) on GAG metabolism in normal and otosclerotic human bone cells as well as its capacity to modulate IL-1α, IL-1β and IL-6 secretion in both populations was analyzed. The amount of radiolabeled GAGs was lower in otosclerotic than in normal bone cells. IL-1α reduced newly synthesized cellular and extracellular GAGs in normal cells, but only those of the cellular compartment in otosclerotic bone cells. It depressed heparan sulphate (HS) more in normal cells and chondroitin sulphate (CS) more in otosclerotic bone cells. The HA/total sulphated GAG ratio was shifted in favour of the latter in otosclerotic cells, whereas the opposite effect was seen after IL-1α treatment. There was little difference in the β-D-glucuronidase levels of the normal and pathological cells, while β-N-acetyl-D-glucosaminidase was significantly increased in otosclerotic bone cells. As the activity of neither enzyme was modified by treatment with IL-1α, the cytokine seems to exert its influences on GAG synthesis rather than on the degradation process. IL-1α, IL-1β and IL-6 secretion was markedly higher in otosclerotic cells. IL-1α modulated the secretion of each interleukin differently, thus resulting in a cytokine cascade that may act in autocrine/paracrine manner on target cells. The authors suggest that changes in the cytokine network may have a specific, yet still unknown, role during normal and pathological osteogenesis.

The cytogenetic follow-up of a case of refractory anemia with excess of blasts (RAEB) that rapidly evolved to acute myeloblastic leukemia (Ml-FAB type) is described. Bone marrow analysis at presentation revealed two chromosomally abnormal clones that shared an interstitial deletion of the long arm of chromosome 5 (5q−) and a terminal deletion of the short arm of chromosome 12 (12p-), but that differed from one another in the localization of a very similar segment of chromosome 17 (i.e. 17q11−12qter) on two clearly distinct karyotypic sites: 2q37 and 17q25. Fourteen percent of the metaphases examined bore the 2q+ marker and 38% the 17q+ marker; the remaining cells had a normal karyotype. A second study carried out 4 months later, at onset of the acute phase, revealed that the clone with normal karyotype had almost completely disappeared and that there had been an inversion in the ratio of the two abnormal cell populations. In the final study, made 1 month before death, the cells with t(2;17) had totally effaced the other clone.

cDNA clones encoding zinc finger motifs were isolated by screening human placenta and T-cell (Peer) cDNA libraries with zinc finger (ZNF) consensus sequences. Unique cDNA clones were mapped in the human genome by rodent-human somatic cell hybrid analysis and in some cases in situ chromosomal hybridization. ZNF 80 mapped to 3p12-3qter, ZNF 7 was previously mapped to 8q24 and is here shown by in situ hybridization and use of appropriate hybrids to map telomeric to the MYC locus. ZNF 79 mapped to 9q34 centromeric to the ABL gene and between a constitutional chromosomal translocation on the centromeric side and the CML specific ABL translocation on the telomeric side. ZNF77 mapped to 19p while ZNF 78L1 (pT3) mapped to 19q. Chromosome 19 carries many ZNF loci and other genes with zinc finger encoding motifs; the pT3 clone additionally detected a locus designated ZNF 78L2, which mapped to chromosome region 1p, most likely in the region 1p32 where the MYCL and JUN loci map.

The finger motif is a tandemly repeated DNA-binding domain recently identified in the primary structure of several eukaryotic transcriptional regulatory proteins. It has been proposed that some members of the finger-gene family are implicated in both normal cell proliferation and differentiation. We isolated several human finger genes by means of hybridization with a finger motif-containing DNA probe. One of these finger genes, HF.10, is expressed at low levels in a variety of human tissues and is down-regulated during the in vitro terminal differentiation of human leukemic myeloid cell lines. By in situ hybridization experiments and analysis of interspecific somatic cell hybrids we mapped the HF.10 gene to 3p21–22, a chromosome region frequently involved in karyotypic rearrangements associated with lung and renal cancer.

The authors treated a total of 82 patients with Ph-positive chronic myelogenous leukemia (CML) with recombinant interferon alpha-2b (IFN α-2b). Sixty-five patients in chronic phase (CP), 28 of whom were untreated and 37 pretreated, and nine patients in accelerated phase (AP), were started on IFN three times a week. Patients in CP were randomized to receive 2 or 5 x 106IU/m , while patients in AP were all given the dose of 5 x 106IU/m2, in addition to concomitant chemotherapy. Patients in CP who were unresponsive to the lower dose were crossed to the higher dose. Of 63 evaluable patients in CP, 43 (68%) responded, 29 (46%) achieved complete hematologic response (CHR), and 14 (22%) achieved partial hematologic response (PHR). The response rate appeared to be significantly influenced by the IFN dose in pretreated patients. Of the nine patients in AP, two attained PHR and one CHR. More recently, eight previously untreated CP cases were submitted to daily IFN administration at doses from 2 to 5 x 106IU/m2. This daily schedule was also applied to patients who had obtained, with the intermittent treatment, a PHR persisting unmodified for six months (nine patients) or an unstable CHR (five patients). Seven of the eight previously untreated patients, and five of the nine PHR patients crossed to daily IFN reached CHR. In the total series of previously untreated patients, the response rate proved to be significantly influenced by the initial risk status. Cytogenetic improvement was seen in 37 of 53 responders (70%) treated for more than 3 months, the median of Ph‘-positive cells declining from 100% to 65% (range 0% to 95%). Complete suppression of Ph’ chromosome was observed in one case. The cytogenetic response was persistent for over 6 months in 21 patients, but the lowest value of Ph' positivity was usually unstable. At a median follow-up of 56 weeks, 23 of 36 (64%) CHR patients remain in continued disease control with IFN. A blastic transformation (BT) occurred in seven of 21 unresponsive patients and in one of the 36 CHR patients. The authors' data confirm that IFN a-2b, especially at daily doses, is effective in inducing clinical and cytogenetic response in a good proportion of patients with CML in the benign phase. Longer follow-ups will define the exact influence of this agent on the natural course of the disease. © 1988 by Grune & Stratton, Inc.

No Abstract.

No abstract

The occurrence of additional chromosomal abnormalities (ACAs) in Philadelphia chromosome (Ph’) positive cells, a phenomenon termed clonal evolution (CE), reflects an increased genetic instability of the leukemic population which progressively acquires a highly malignant phenotype. Few data are available regarding the therapeutic effects exerted by imatinib mesylate on Ph’ cells bearing ACAs. Herein, we report the activity of imatinib mesylate employed as single agent or in combination with recombinant interferon (rIFN)a2 in inducing cytogenetic and molecular responses in 14 CML patients (F:M=5:9, median age: 58 years) with CE already present at diagnosis (5) or occurring later during the course of the disease (9) as the sole sign of accelerated phase (AP) (12) or associated with other AP features (2). Overall, the analysis of ACA rate showed the presence of i(Ph’) in 4, +8 in 3, -Y in 3, variant translocation in 2 (t(9;19;22) and t(3;9;22)), -17 in 1, 17p- in 1, 3p- in 1, 22q- in 1, and additional translocations involving chromosomes other than 9 and 22 in 3 other instances. Of note, the two cases with variant translocation showed at diagnosis the classical t(9;22). The t(9;19;22) or t(3;9;22), which was documented later in the course of the disease was therefore a proven second event. Imatinib was given at 400mg po/daily in all, but 2 patients with signs of AP in addition to CE who received a daily dose of 600 mg. rIFNa2 was given at a daily dose of 1 to 3x106UI, according to patient’s tolerance. An overall cytogenetic remission rate of 64% (9/14) was documented in response to imatinib mesylate employed as single agent (6/10) or in combination with rIFNa2 (3/4). Suppression of the ACA population was documented within an interval of 3–14 months from the beginning of imatinib treatment and it was timely coincident with the occurrence of CCR in 6 patients, irrespective of the entity of the ACA population in the context of the entire Ph’ positive one. In 2 of them, a “biphasic” response to imatinib was observed. Initially, suppression of i(Ph’) and t(9;19;22) with re-emergence of the original t(9;22) hemopoiesis was documented 4 and 7 months after treatment initiation, respectively, and then CCR occurred after further 10 and 9 months of treatment, respectively. In the last patient, a mixture of non-clonal and clonal +8 Ph’ negative hemopoiesis was noted at the time when CCR was documented. All 9 CCR patients tested negative for BCR/ABL gene at FISH analysis. Six of the 9 CCR obtained a major or complete molecular remission 3–29 months from treatment initiation. In 3 additional patients, who never achieved a CCR, suppression of the secondary clones was documented. In 1 case, the clone bearing 17p- was no more detected from the 6th to the 42nd month of therapy, when a new clone showing the i(17q) was recorded. In the other 2 cases, the leukemic population bearing the 3q- and 22q- or -17 marker was suppressed 6 and 3 months after the beginning of imatinib mesylate treatment, respectively. After a median follow up of 70 months, 8 patients are still alive and in continuous CCR. Such a long follow up, along with the rate, quality and duration of the responses observed, suggest that CE does not compromise the therapeutic activity of imatinib alone or combined with rIFNa in CML.

The occurrence of additional chromosomal abnormalities (ACAs) in Philadelphia chromosome (Ph') positive cells, a phenomenon termed clonal evolution (CE), reflects an increased genetic instability of the leukemic population which progressively acquires a highly malignant phenotype. Few data are available regarding the therapeutic effects exerted by imatinib mesylate on Ph' cells bearing ACAs. Herein, we report the activity of imatinib mesylate employed as single agent or in combination with recombinant interferon (rIFN)a2 in inducing cytogenetic and molecular responses in 14 CML patients (F:M=5:9, median age: 58 years) with CE already present at diagnosis (5) or occurring later during the course of the disease (9) as the sole sign of accelerated phase (AP) (12) or associated with other AP features (2). Overall, the analysis of ACA rate showed the presence of i(Ph') in 4, +8 in 3, -Y in 3, variant translocation in 2 (t(9;19;22) and t(3;9;22)), -17 in 1, 17p- in 1, 3p- in 1, 22q- in 1, and additional translocations involving chromosomes other than 9 and 22 in 3 other instances. Of note, the two cases with variant translocation showed at diagnosis the classical t(9;22). The t(9;19;22) or t(3;9;22), which was documented later in the course of the disease was therefore a proven second event. Imatinib was given at 400mg po/daily in all, but 2 patients with signs of AP in addition to CE who received a daily dose of 600 mg. rIFNa2 was given at a daily dose of 1 to 3x106UI, according to patient's tolerance. An overall cytogenetic remission rate of 64% (9/14) was documented in response to imatinib mesylate employed as single agent (6/10) or in combination with rIFNa2 (3/4). Suppression of the ACA population was documented within an interval of 3–14 months from the beginning of imatinib treatment and it was timely coincident with the occurrence of CCR in 6 patients, irrespective of the entity of the ACA population in the context of the entire Ph' positive one. In 2 of them, a “biphasic” response to imatinib was observed. Initially, suppression of i(Ph') and t(9;19;22) with re-emergence of the original t(9;22) hemopoiesis was documented 4 and 7 months after treatment initiation, respectively, and then CCR occurred after further 10 and 9 months of treatment, respectively. In the last patient, a mixture of non-clonal and clonal +8 Ph' negative hemopoiesis was noted at the time when CCR was documented. All 9 CCR patients tested negative for BCR/ABL gene at FISH analysis. Six of the 9 CCR obtained a major or complete molecular remission 3–29 months from treatment initiation. In 3 additional patients, who never achieved a CCR, suppression of the secondary clones was documented. In 1 case, the clone bearing 17p- was no more detected from the 6thto the 42ndmonth of therapy, when a new clone showing the i(17q) was recorded. In the other 2 cases, the leukemic population bearing the 3q- and 22q- or -17 marker was suppressed 6 and 3 months after the beginning of imatinib mesylate treatment, respectively. After a median follow up of 70 months, 8 patients are still alive and in continuous CCR. Such a long follow up, along with the rate, quality and duration of the responses observed, suggest that CE does not compromise the therapeutic activity of imatinib alone or combined with rIFNa in CML.

The rate of complete karyotypic remission (CKR) and complete molecular remission (undetectable BCR/ABL specific transcript by real-time quantitative-PCR (Gabert J et al. Leukemia: 2003; 17: 2318–57) and NESTED/real-time-PCR (Guo JQ et al.; Leukemia: 2002; 15:2447–53)) to various therapies was analysed in 35, of the 114 CML patients observed in our institution, who showed cytogenetic clonal evolution. Overall, 52 karyotypic abnormalities additional to Ph’ were documented at the time of diagnosis (early clonal evolution) or late in the course of the disease (late clonal evolution). Seven of 12 patients with early clonal evolution achieved one (5 pts) or more (2 pts) CKRs, for a total number of 9, in response to IFNα (2 CKRs), IFNα plus ARA-C (1 CKR), imatinib (5 CKRs) or imatinib-IFNα combination (1 CKRs). Six of the 23 patients with late clonal evolution achieved one or more CKRs during the course of their disease after treatment with IFNα (2), IFNa plus ARA-C (1) or imatinib (5). In addition, 7 of the 13 CK responders reached a complete molecular remission in response to IFNa (1), IFNa plus ARA-C (1) imatinib (1), or imatinib plus IFNa (4). The 5 complete molecular remissions documented in patients with early clonal evolution, were observed following treatment with IFNα plus ARA-C (1), imatinib (1), or imatinib plus IFNα (3). Two patients with late clonal evolution obtained complete molecular remission in response to IFNα and imatinib plus IFN therapy, respectively. Cytogenetic and molecular response rates were higher in patients with early (58%, 57%) than in patients with late secondary clones (26%, 33%), while there were no significant differences in time to achieve cytogenetic (2–20 months vs 1–25 months) or molecular (3–26 months vs 13–52 months) remission and the duration of response (11 months, range 4–42 vs 10 months, range 7–46 and 2–19 vs 3–13+ months, respectively) between the two groups. Finally, no relationship was evident between the type of additional karyotypic anomalies and sensitivity to treatment. In fact, clones bearing complex translocations (2), isoPh’ (2), trisomy 8 (2), translocations other than Ph’ (2), partial chromosome monosomy (11-q, 1) or loss of chromosome Y alone (1) or associated with other anomalies (−6 and −14 or −21 or isoPh’) (3) all responded to therapy. In conclusion, although the small sample size, these findings suggest that the presence of karyotypic abnormalities additional to Ph’ in CML is not associated to disease resistance to therapy.

Fifty-two kariotypic abnormalities additional to Philadelphia chromosome (Ph’) were documented at the time of diagnosis or during the course of the disease in 114 CML patients. Five were evaluated only once at the time of diagnosis, while 109 were repeatedly studied overtime during a median period of 138 (1–275)months. A clonal evolution was already present at diagnosis (early clonal evolution) in 12, while in 23 patients, it was documented in the course of the disease (late clonal evolution). Simultaneous appearence of multiple chromosome abnormalities (4/12 and 4/23) and a tendency to acquire further alterations (5/12 and 2/23, respectively) were associate with both early and late secondary clones. Partial or complete chromosome losses (-1, -6, 6q-, 7p-, 11q-, 12p-, -14, -16, -17,-18, -21) other than -Y (11 events), + 8 (9) and loss of Y chromosome (8) were the most commonly cytogenetic abnormalities, irrespective of the time of their occurrence. Translocations other than Ph’ (5 events), complex Ph’ translocations (5), duplication of Ph’ (5), single iso(Ph’) (4), duplication of iso(Ph’) (1) or iso17q (2) were found with a decreasing frequency. The type of kariotypic alteration varied with the time of onset. In fact, partial or complete chromosome monosomies and duplication of iso(Ph’) never represented early, but only late events. Duplication of Ph’ was a late (4/5) rather than an early event (1/5). Similarly, the presence of the iso(Ph’) was documented either in early or late clonal evolution in 1 and 3 instances respectively. Furthermore, +8 developed several months after diagnosis in all, but 2 cases. Complex Ph’ translocations or loss of Y chromosome do not correlate with any specific phase of the leukemic course. Cytogenetic abnormalities neither correlated with Sokal risk nor with the type of break-points (b3a2,b2a2) documented at diagnosis. However all 3 clones bearing p230 chimeric protein showed early cytogenetic evolution. Secondary clones seem to have different impact on disease history. The occurrence of partial or complete deletions, the iso17q and the late, but not early, loss of Y chromosome were all predictive of a rapid disease progression. Early secondary clones with additional translocations had no effect on the chronic behaviour of the disease, while such clones were associated with a rapid progression (2/3) to blastic phase when they occurred during the course of disease. The presence of +8 as the sole cytogenetic abnormality and complex translocations, irrespective of the time of their onset, did not correlate with leukemia evolution. Finally,simple Ph’ duplication of or single and double iso(Ph’), when not associated with other cytogenetic abnormalities, did not correlate with an aggressive course, even if it can be assumed that these cytogenetic events are responsible for an increased concentration of the p210 oncoprotein. In conclusion, regardless of whether bcr/abl has a direct or an indirect role in promoting genomic instability, in our cohort of CML patients additional non random chromosomal abnormalities frequently characterize both early and late phase of the disease and some of them may represent crucial steps in development and progression of leukemia.

Introduction Progressive improvement has been observed in CKR and survival of CML patients (pts) in response to interferon (IFN)a-based regimens, or imatinib. The purpose of this study is the evaluation of: rate of, time to and duration of CKR in accordance to first line therapy employed and Sokal score; impact on overall survival of CKR, and Sokal score, separately considered or combined together. Patients. 109Ph’+ and 5Ph’-, (BCR-ABL positive), CML pts were treated at diagnosis with allogeneic transplantation (3 pts), hydroxyurea (HU) (19 pts), INFa (51pts, G1), INFa associated with ARA-C (20 pts, G2), imatinib alone (18 pts, G3), or imatinib combined with INFa (3 pts, G4). INFa was employed as second line therapy in 12 pts initially treated with hydroxyurea (G5), while INFa/ARA-C combination or imatinib alone was given to 24 (G6) and 23 (G7) pts with de novo or acquired resistance or intolerance to INFa. Third line therapy, consisting of the combination of imatinib with IFNa, was employed in 11 (G8) pts with no CKR (5 pts) or in complete cytogenetic, but not in molecular remission (6 pts). Results. 40 of 94 Ph’+ evaluable non-allotransplanted pts obtained one or more (overall 47) CKRs to INFa-based regimens or imatinib. CKR rate, median time to CKR and response duration are shown in table 1. In the analysis according to Sokal score 82/94 pts, with complete prognostic data at diagnosis, were included. The percentage of responders was higher in the low compared to the non-low Sokal risk group (57% vs. 31%). Irrespective of the treatment, median duration value of the first CKR was also better in the former [18+mths(1–64)] than in the latter group [6mths(2–54)] with 16 vs.4 pts still in first or subsequent remission. Overall survival for CKRs was 68+mths(5–275) vs. 52mths(5–270) for CKRs with 35 vs. 6 pts still alive respectively. Overall survival according to Sokal score at diagnosis was 61+ mths(5–275) for low vs 53mths(5–212) for non-low risk patients. The impact on survival of CKR and Sokal risk were then analyzed simultaneously. The median survival of 27 CKRs and 20 not CKRs with low Sokal risk were 61+mths(5–275)and 63 mths(14–270) respectively as compared to 73+mths(11–212) of 11 CKRs and 36mths(5–139) of 24 not CKRs with unfavourable characteristics at diagnosis. The number of patients still alive in these 4 groups were 24/27, 3/20, 8/11, 3/24 respectively at the time when this analysis was performed. Conclusions. The present data not only confirm the effectiveness of imatinib-over the INFa-based regimens in inducing CKR, but also suggest that response to treatment may be better than Sokal risk in predicting patient survival. Rate,time to and duration of CKR according to treatment PT Group CKR rate(%) time to CKR( months) Duration of CKR (months) N° of pts in cCKR Median Range Median Range G1 17 16 (3–28) 6 (2–53) 0 G2 30 11 (3–24) 15,5 (2–33) 0 G3 85 6 (2–14) 10+ (1–44) 11 G4 100 4 (4–5) 48 (2–50) 1 G5 0 0 G6 9 9,5 (7–12) 40,5 (17–64) 1 G7 54 9 (4–38) 21,5+ (3–54) 6 G8 80 3 (2–4) 26+ (4–48) 9

The rate of complete karyotypic remission (CKR) and complete molecular remission (undetectable BCR/ABL specific transcript by real-time quantitative-PCR (Gabert J et al. Leukemia : 2003; 17: 2318–57) and NESTED/real-time-PCR (Guo JQ et al.; Leukemia : 2002; 15:2447–53)) to various therapies was analysed in 35, of the 114 CML patients observed in our institution, who showed cytogenetic clonal evolution. Overall, 52 karyotypic abnormalities additional to Ph' were documented at the time of diagnosis (early clonal evolution) or late in the course of the disease (late clonal evolution). Seven of 12 patients with early clonal evolution achieved one (5 pts) or more (2 pts) CKRs, for a total number of 9, in response to IFNα (2 CKRs), IFNα plus ARA-C (1 CKR), imatinib (5 CKRs) or imatinib-IFNα combination (1 CKRs). Six of the 23 patients with late clonal evolution achieved one or more CKRs during the course of their disease after treatment with IFNα (2), IFNa plus ARA-C (1) or imatinib (5). In addition, 7 of the 13 CK responders reached a complete molecular remission in response to IFNa (1), IFNa plus ARA-C (1) imatinib (1), or imatinib plus IFNa (4). The 5 complete molecular remissions documented in patients with early clonal evolution, were observed following treatment with IFNα plus ARA-C (1), imatinib (1), or imatinib plus IFNα (3). Two patients with late clonal evolution obtained complete molecular remission in response to IFNα and imatinib plus IFN therapy, respectively. Cytogenetic and molecular response rates were higher in patients with early (58%, 57%) than in patients with late secondary clones (26%, 33%), while there were no significant differences in time to achieve cytogenetic (2–20 months vs 1–25 months) or molecular (3–26 months vs 13–52 months) remission and the duration of response (11 months, range 4–42 vs 10 months, range 7–46 and 2–19 vs 3–13+ months, respectively) between the two groups. Finally, no relationship was evident between the type of additional karyotypic anomalies and sensitivity to treatment. In fact, clones bearing complex translocations (2), isoPh' (2), trisomy 8 (2), translocations other than Ph' (2), partial chromosome monosomy (11-q, 1) or loss of chromosome Y alone (1) or associated with other anomalies (−6 and −14 or −21 or isoPh') (3) all responded to therapy. In conclusion, although the small sample size, these findings suggest that the presence of karyotypic abnormalities additional to Ph' in CML is not associated to disease resistance to therapy.

Fifty-two kariotypic abnormalities additional to Philadelphia chromosome (Ph') were documented at the time of diagnosis or during the course of the disease in 114 CML patients. Five were evaluated only once at the time of diagnosis, while 109 were repeatedly studied overtime during a median period of 138 (1–275)months. A clonal evolution was already present at diagnosis (early clonal evolution) in 12, while in 23 patients, it was documented in the course of the disease (late clonal evolution). Simultaneous appearence of multiple chromosome abnormalities (4/12 and 4/23) and a tendency to acquire further alterations (5/12 and 2/23, respectively) were associate with both early and late secondary clones. Partial or complete chromosome losses (-1, -6, 6q-, 7p-, 11q-, 12p-, -14, -16, -17,-18, -21) other than -Y (11 events), + 8 (9) and loss of Y chromosome (8) were the most commonly cytogenetic abnormalities, irrespective of the time of their occurrence. Translocations other than Ph' (5 events), complex Ph' translocations (5), duplication of Ph' (5), single iso(Ph') (4), duplication of iso(Ph') (1) or iso17q (2) were found with a decreasing frequency. The type of kariotypic alteration varied with the time of onset. In fact, partial or complete chromosome monosomies and duplication of iso(Ph') never represented early, but only late events. Duplication of Ph' was a late (4/5) rather than an early event (1/5). Similarly, the presence of the iso(Ph') was documented either in early or late clonal evolution in 1 and 3 instances respectively. Furthermore, +8 developed several months after diagnosis in all, but 2 cases. Complex Ph' translocations or loss of Y chromosome do not correlate with any specific phase of the leukemic course. Cytogenetic abnormalities neither correlated with Sokal risk nor with the type of break-points (b3a2,b2a2) documented at diagnosis. However all 3 clones bearing p230 chimeric protein showed early cytogenetic evolution. Secondary clones seem to have different impact on disease history. The occurrence of partial or complete deletions, the iso17q and the late, but not early, loss of Y chromosome were all predictive of a rapid disease progression. Early secondary clones with additional translocations had no effect on the chronic behaviour of the disease, while such clones were associated with a rapid progression (2/3) to blastic phase when they occurred during the course of disease. The presence of +8 as the sole cytogenetic abnormality and complex translocations, irrespective of the time of their onset, did not correlate with leukemia evolution. Finally,simple Ph' duplication of or single and double iso(Ph'), when not associated with other cytogenetic abnormalities, did not correlate with an aggressive course, even if it can be assumed that these cytogenetic events are responsible for an increased concentration of the p210 oncoprotein. In conclusion, regardless of whether bcr/abl has a direct or an indirect role in promoting genomic instability, in our cohort of CML patients additional non random chromosomal abnormalities frequently characterize both early and late phase of the disease and some of them may represent crucial steps in development and progression of leukemia.

Introduction Progressive improvement has been observed in CKR and survival of CML patients (pts) in response to interferon (IFN)a-based regimens, or imatinib. The purpose of this study is the evaluation of:

Introduction: Continous improving results have been obtained during last two decades in the control of Ph’positive chronic myeloid leukemia(CML). However the final goal of molecular remission remains difficult to obtain even in the STI age. Aims : Evaluation of the rate of molecular response to IFNα,IFNα based treatment,to STI or to STI-INFα combination was analized in 100 consecutive Ph+ CML patients observed in a single Institution over a period of 20 years. Patients, Methods and Results All patients were treated at the time of diagnosis (87) or late (13) during the course of their disease. Distribution according to treatment was: INFα,63pts (late or early:13,50);INFα-ARA-C combination,20pts;STI,14 pts;STI-INFα association, 3 pts. Two pts, both initially assigned to INFα-ARA-C combination, were crossed-over to STI, one because relapsing off-therapy after a long lasting continous (25 mths) molecular remission and the other in cytogenetic response because intolerant to the initial treatment. In addition, other 3 pts patients, with persistent complete cytogenetic, but not molecular remission to INFα or INFα-ARA-C combination were subsequentially trated with the STI-IFNα association. At present,99/100 pts are evaluable. The median times of follow-up for the entire group and form the different treatment subgroups are: late IFNα 154 months(42–263); early IFNα, 71 months(1–197); IFNα-ARA-C, 61 months(5–203); STI- IFNα,78 mths(11–47), STI,31 mths(3–41). A complete kariotypic remission(CKR) was observed in 15/63 IFNα treated pts, in 10/20 IFNα-ARA-C pts group, in 10/13 cases of STI group and in 3 /3 pts who received STI-IFNα. A molecular response(RT-nested PCR, JQ Guo, Leukemia: 2002,15,2447–53) was observed in 4/15,2/10,5/10 and in 2/3 CKR pts initially trated with the different modalities listed above. Response was confirmed from 2 to 7 consecutive or not consecutive times in the 2/4 cases responsive to INFα, in the 2 cases responsive to INFα-ARA-C combination,4/5cases responsive to STI and in 2/3 cases responsive to STI-IFNα association. The 2nd and the 3rd molecular remission to STI were obtained in the patient molecularly and cytogenetically relapsed off-therapy and, for the first time from the diagnosis, in the other patient in CKR to IFNα-ARA-C combination and crossed to STI treatment. Furthermore, all 3 cases, in CKR, but not molecular response to other treatments at the time of cross-over to STI-IFNα combination, achieved a persistent (in 2 to 3 tests over a period ranging from 6+ to 12+ mths) molecular remission. The first interval between the start of the treatment and the first molecular response varied from 12 to 52, from 3 to 22, from 11 to 24, from 5 to 11 mths in the groups initially treated with IFNα, IFNα-ARA-C, STI or STI-IFNα respectively. The 2 pts, crossed-over to STI alone, both, obtained a response after 29 mths of therapy. In addition in the 3 pts crossed-over to STI-IFNα therapy, the molecular response was obtained after 14,23 and 25 mths from the start of last treatment. Conclusion It is not possible to achieve any conclusion regarding the treatment effect on molecular response duration because of the different length of follow-up of various groups of patients. However in responsive patients to IFN alone or combined to ARA-C or STI, consecutive negative RT-PCR tests were observed more frequently than in patients receving STI alone.

Introduction:

A phenotypically otherwise normal homosexual man with a 46,XY,inv(2)(q21q33) karyotype inherited from his mother is described. The breakpoints were different from those observed in the only other case of familial paracentric inversion of chromosome 2 reported in the literature, but in our case they seem to correspond to constitutive and aphidicolin-induced fragile sites.