Copies Of 1q Research Articles

Additional copies of 1q negatively impact the outcome of multiple myeloma patients and induce transcriptomic deregulation in malignant plasma cells

Additional copies of chromosome 1 long arm (1q) are frequently found in multiple myeloma (MM) and predict high-risk disease. Available data suggest a different outcome and biology of patients with amplification (Amp1q, ≥4 copies of 1q) vs. gain (Gain1q, 3 copies of 1q) of 1q. We evaluated the impact of Amp1q/Gain1q on the outcome of newly diagnosed MM patients enrolled in the FORTE trial (NCT02203643). Among 400 patients with available 1q data, 52 (13%) had Amp1q and 129 (32%) Gain1q. After a median follow-up of 62 months, median progression-free survival (PFS) was 21.2 months in the Amp1q group, 54.9 months in Gain1q, and not reached (NR) in Normal 1q. PFS was significantly hampered by the presence of Amp1q (HR 3.34 vs. Normal 1q, P < 0.0001; HR 1.99 vs. Gain1q, P = 0.0008). Patients with Gain1q had also a significantly shorter PFS compared with Normal 1q (HR 1.68, P = 0.0031). Concomitant poor prognostic factors or the failure to achieve MRD negativity predicted a median PFS < 12 months in Amp1q patients. Carfilzomib–lenalidomide–dexamethasone plus autologous stem cell transplantation treatment improved the adverse effect of Gain1q but not Amp1q. Transcriptomic data showed that additional 1q copies were associated with deregulation in apoptosis signaling, p38 MAPK signaling, and Myc-related genes.

Outcomes of patients with multiple myeloma and 1q gain/amplification receiving autologous hematopoietic stem cell transplant: the MD Anderson cancer center experience

The prognostic impact of additional copies of chromosome 1q (1q + ) on outcomes of newly-diagnosed multiple myeloma (NDMM) patients undergoing autologous transplantation (autoSCT) is unclear. We conducted a retrospective single-center analysis of NDMM patients with 1q21 gain/amplification (3 or ≥4 copies of 1q, respectively) that received autoSCT between 2008–2018. 213 patients were included (79% 1q gain; 21% 1q amplification). The most commonly used induction regimen was bortezomib, lenalidomide, and dexamethasone (41%). At day100 post-autoSCT and at best post-transplant response, 78% and 87% of patients achieved ≥VGPR, and 38% and 50% achieved MRD-negative ≥VGPR, respectively. Median PFS and OS for the entire cohort were 35.5 months and 81.4 months, respectively. On multivariable assessment for PFS, MRD negative ≥VGPR before autoSCT (HR 0.52, p = 0.013) was associated with superior PFS, whereas 1q amplification was associated with inferior PFS (2.03, p = 0.003). On multivariate analysis for OS, achieving MRD negative ≥VGPR at best post-transplant response was associated with superior survival (0.29, p < 0.001), whereas R-ISS III and concomitant del17p or t(4:14) were associated with inferior survival (6.95, p = 0.030, 2.33, p = 0.023 and 3.00, p = 0.047, respectively). In conclusion, patients with 1q+ NDMM, especially 1q amplification, have inferior survival outcomes compared to standard-risk disease after upfront autoSCT, though outcomes are better than other high-risk cytogenetic abnormalities.

Outcomes of Patients with Multiple Myeloma and 1q Gain/Amplification Receiving Autologous Hematopoietic Stem Cell Transplant: The MD Anderson Cancer Center Experience

Background: There is contradictory data regarding the prognostic impact of additional copy numbers of chromosome 1q (1q+) on the outcomes of newly diagnosed multiple myeloma (NDMM) patients, and there is scarce data in the context of autologous stem cell transplantation (autoSCT). In this report, we studied outcomes of NDMM patients with 1q+ who received induction with contemporary anti-myeloma agents, followed by autoSCT and post-transplant maintenance therapy. Methods: We conducted a retrospective single-center chart review analysis of adult NDMM patients with 1q21 gain or amplification (3 or ≥4 copies of 1q, respectively; 1q+) detected by fluorescence in situ hybridization (FISH) that received autoSCT between 2008-2018. Progression-free survival (PFS) and overall survival (OS) were the primary endpoints. Results: 213 NDMM patients with 1q+ were included in the analysis, with a median age of 62.5 years and 53% were male. Overall, 169 (79%) patients had 1q gain, while 44 (21%) patients had 1q amplification. The most commonly used induction and conditioning regimens were bortezomib, lenalidomide, and dexamethasone (VRD) (41%) and melphalan (77%), respectively (Table 1). At day 100 after autoSCT and at best post-transplant response, 78% and 87% of patients achieved ≥VGPR, 34% and 56% achieved CR, 38% and 50% achieved MRD negative ≥VGPR, respectively. The median PFS and OS for the entire cohort were 35.5 months and 81.4 months, respectively. 1q amplification was associated with inferior PFS compared to 1q gain (HR=2.03, 95% CI 1.36-3.03, p&amp;lt;0.001; Figure 1). On multivariable assessment (MVA) for PFS, MRD negative &amp;gt;VGPR before autoSCT and at day 100 post-transplant [(HR=0.56, 95% CI 0.36-0.86, p=0.009) and (HR=0.64, 95% CI 0.44-0.94, p=0.022), respectively] were associated with better PFS, whereas 1q amplification was associated with inferior PFS (HR=1.94, 95% CI 1.29-2.92, p=0.001). On MVA for OS, R-ISS stage III (HR=4.08, 95% CI 1.07-15.50, p=0.039) was associated with inferior OS, whereas achieving MRD negative &amp;gt;VGPR at best post-transplant response was associated with superior OS (HR=0.45, 95% CI 0.24-0.85, p=0.014). Notably, the percentage of cells with 1q+ was not associated with PFS nor with OS, both when evaluated as a continuous variable and when evaluated as a categorical variable using thresholds of either 30% or 50%. The presence of additional high-risk cytogenetic abnormalities did not adversely affect survival outcomes. Conclusions: Patients with NDMM and 1q+, especially 1q amplification, have poor survival outcomes, despite the use of contemporary induction regimens, upfront autoSCT and post-transplant maintenance. These patients may benefit from novel treatment modalities, such as CAR-T and bispecific antibodies, earlier in their disease course.

Mitotic recombinatory evolution in acute leukemia

A cohort of leukemia cases is presented with ancillary testing that includes microarray studies, karyotyping, FISH, and RNA sequencing to illustrate clonal evolution. Common evolution etiology with each case is apparent homologous mitotic recombination (HMR). The cohort includes: four cases of Pre B-cell acute lymphoblastic leukemia (B-ALL) with a single translocation derivative (19)t(1;19)(q23.3;p13.3), an acute myelogenous leukemia (AML) case with a paracentric inversion of 11q13.3q23 in both homologues confirmed as a rare KMT2A-MAML2 gene fusion, and a transplant patient in AML relapse with a t(6;11)(6q27;q23) and evolution to an additional derivative 6 chromosome. The PBX1-TCF3 fusion in the t(1;19) B-ALL subgroup has long been associated with clones that show either the balanced translocation (∼25%) or the unbalanced single derivative 19 (∼75%). Evidence from the CMAs and FISH is consistent with HMR initiating at either the PBX1 translocation breakpoint or at a more proximal long arm site that mediates the evolution to the unbalanced form. This is contrary to the previous assumptions of either nondisjunction duplication of the normal homologue with loss of the translocation derivative 1 or an original trisomy 1 that loses the translocation derivative 1. Relapse from an unrelated transplant donor created unique allele dosage ratios in the microarray of the AML patient with the t(6;11) KMT2A-AFDN fusion. An HMR-based evolution initiation site proximal to the 6q27 AFDN fusion gene is evident in the microarray of chromosome 6, the known oncogenic fusion derivative. The HMR selection driver in both AML cases is very likely associated with the DNA doubling of the oncogenic fusions in 6q and 11q, respectively. Since the oncogenic derivatives in the 1;19 cases are clearly the retained derivative 19, selection for the HMR clonal evolution in 1q is apparently based on the known proliferative advantage of extra copies of 1q in B-ALL and other malignancies. Although selection-based HMR can effectively initiate at any site proximal to a driver gene fusion, it appears that the translocation breaksite is common for many translocations. In addition, evidence from HMR evolution related distal 11q mutations, numerous unbalanced CCND1/IGH translocations, and the double MAML2/KMT2A presented in this study suggest that a recombinatorial "hot spot" exists near the CCND1 gene in many rearrangements or mutations within 11q.

The prognostic role of 1q21 gain/amplification in newly diagnosed multiple myeloma: The faster, the worse.

The prognostic value of additional copies of chromosome 1q (1q gain/amplification [amp]) in multiple myeloma (MM) remains controversial. In the meantime, the kinetics of the response to MM therapy has long been an area of debate. Few studies have pointed out the relationship of response kinetics with cytogenetic abnormalities (CAs) in MM. The authors retrospectively analyzed the data of 1068 real-world newly diagnosed MM patients from a Chinese national medical center. Overall, 405 (51.9%) patients had 1q gain/amp, with aggressive clinical characteristics and significant inferior survival. The variation in copy number (CN) of 1q (CN=3 or CN>3) had no significant impact on the survival of MM patients with 1q abnormalities. No difference was found in the outcome of 1q gain/amp patients treated with doublet or triplet regimens. Upfront autologous stem cell transplantation could eliminate the adverse prognostic effect of 1q gain but not 1q amp. The duration from diagnosis to the first time achieving very good partial response (VGPR) or better was significantly shorter in patients with 1q gain/amp (77days vs. 100days, p=.001). Finally, multifactor regression analysis was performed to construct a new risk stratification model in MM patients with 1q gain/amp, which was validated in the Multiple Myeloma Research Foundation CoMMpass study cohort and worked better than the Revised International Staging System and Second Revision of the International Staging System (Harrell's concordance index: 0.631 vs. 0.598 and 0.537). In the setting of novel therapy, 1q gain/amp still acts as an independent adverse prognostic factor. Patients with 1q gain/amp achieved VGPR rapidly but had inferior survival.

Structural Heterogeneity of Chromosome 1q Drives Outcome in Newly Diagnoses Myeloma

Background: The prognostic importance of 1q gain, present in 30% of cases, has led to it being extensively studied and incorporated into prognostic systems. Amplification (>3 copies) of 1q is seen in 10% of cases and has a greater degree of specificity for poor outcome than gain alone. Despite this, there is ongoing debate about its value, likely due to differences in the nature and penetrance of deregulated drivers. To gain insight into this we dissected the structural changes seen at the 1q level and integrated them with expression and clinical data to identify potential drivers and gain insight into progression mechanisms. Methods We analyzed data derived from 1,154 CoMMpass trial patients including 972 NDMM patients with whole exome for mutations, and 752 whole genomes for copy number (CNA), translocations, and complex rearrangements. Using GISTIC 2.0 and PFC we identified hotspots of CNA. This information was analyzed in conjunction with the RNA-seq data derived from 643 patients to determine the aberrant transcriptional landscape of 1q. Results: We identified nine regions of gain (numbered G1 to G9), eight of them being on 1q. Several potential oncogenes such as BCL9 (G2), MCL1 (G3), SLAMF7 (G5), POU2F1 (G6), and BTG2 (G8) lay within these regions of gain. Of note, other previously identified drivers lay outside specific hotspots: ANP32E (306 kb downstream of G3) CKS1B and ADAR1 (1209 kb ( and 1561 kb away from G4, respectively), ATF6 (941 kb from G5), and PBX1 (2313 kb upstream of G6). These data suggest that although important, these hotpots do not drive the entire spectrum of genes upregulated as a consequence of gain 1q. We noted that 302 samples (40%) had at least one gained region with 80% (241/302) having gain of 7-9 regions; 69% (207/302) had gain G2-G9. Each of the gained regions occurred at a similar frequency (G2-G9 range 79-87%) with the exception of G1 located at the sub-telomeric border of 1p seen only in 6% (18/302) of patients. There were 219 cases with gain of G2-G9. These results show two broad groups of gain 1q; whole arm gains and a group with focal gain. Patients with whole arm gain had a worse PFS (HR 1.3 (95% CI 1.01-1.6) and OS (HR 1.6 (95% CI 1.15-2.2) than patients with no 1q gain; the outcome for patients focal events was neutral. Whole arm gains strongly associated with a complex genomic background including with t(4;14) (corr=0.10, BF=1.06), loss of acrocentric chromosomes (del(13q) corr=0.23, BF=118623 and del(14q) corr=0.12, BF=2.9) and other deletions (del(4p) corr=0.11, BF=1.89, del(16p) corr=0.12, BF=2.3) and negatively correlated to trisomies and t(11;14) (corr=0.16, BF=184). Interestingly whole arm gains had significantly shorter telomeres and a higher mean age, and patients with critically short tumor telomeres were over-represented (Corr=0.11, BF=1.2). Common associations with CKS1B gain are mostly found in the whole arm gains group. Amp(1q) was over represented in the whole arm gain group (87%) and associated with the worst outcome (data not shown). Similarly, focal events strongly associated with templated insertion (corr=0.34, BF=6e14), MYC translocation (corr=0.24, BF=3034538), and HRD (corr=0.13, BF=4.9). To address expression changes in focal changes we compared patients with no 1q gain (n=297) with patients with a focal event (n=56); we identified 20 genes of which the majority (n=12) were located on chromosome 1 and related to protein handling. We compared patients with no evidence of 1q gain with patients with a whole arm gain (n= 157); the number of deregulated genes was higher both on 1q and overall (n=585 and n=2409 respectively) suggesting that generalized gene deregulation is part of a more complex genome wide mechanism. Interestingly, 101 were transcription factors and included upregulation of BACH2, NR5A1, MYBL1. GLMP, and E2F2 and downregulation of PAX5, SMAD1, TBX21, SIX4, EGR3, and GF1 most of them being involved in histone methylation, intracellular signaling, and regulation of transcription. Conclusions: Our data add precision by which 1q is associated with an adverse outcome and an aggressive disease phenotype by separating out focal gains, that occur on a favorable background and associate with a good outcome and those with arm-length gains, associated with high-risk disease features, deregulation of multiple transcriptional programs, increased metabolism and aggressive clinical behavior. Figure 1View largeDownload PPTFigure 1View largeDownload PPT Close modal

Presentation and Impact of Double and Triple hit Cytogenetics in Patients With Multiple Myeloma in the Real World

The presence of 2 or more high-risk cytogenetic abnormalities on FISH characterizes patients as Double/Triple Hit Myeloma (DH/THM). This entity has poorer outcomes, though data from the real-world setting is scarce. We retrospectively analyzed all MM patients presenting to our hospital over 3 years to study the clinical characteristics and outcomes with DH/THM. Fifty-five (19.2%) of 285 newly-diagnosed MM patients had DH/THM. The most common cytogenetic abnormality was increased copies of chromosome 1q (53 patients, 96.4%)- 22 patients had 3 copies, and 31 patients had 4 or more copies. The most common cytogenetic combination was IgH-FGFR3 translocation, increasing 1q copy number (28 patients; 50.9%). The disease was characterized by aggressive presentation, with 9 patients (16.4%) dying in the first month after diagnosis, and 7 patients (12.7%) dying in the second month. Only 22 patients (40%) achieved a VGPR or better with therapy. The Median EFS and OS for the cohort were 8 months and thirteen months, respectively. On multi-variate analysis, renal failure and plasma cell leukemia were associated with increased mortality risk, while achievement of VGPR or better was associated with better survival. Our findings suggest that multiple myeloma patients with double/triple hit cytogenetics on FISH have aggressive presentations, high early mortality, and poor outcomes. These patients may benefit from more aggressive regimens and better supportive care.

Early Mortality and Aggressive Presentation Lead to Poor Outcomes in Patients with Newly Diagnosed Double Hit and Triple Hit Multiple Myeloma

INTRODUCTIONmSMART classifies high-risk Multiple Myeloma patients into Double Hit and Triple Hit Myeloma (DH/THM) on the basis of the number of high-risk cytogenetic abnormalities detected. The outcome of these patients with the use of proteasome inhibitors and IMiDs with upfront Auto HSCT is not known in the real-world setting. We retrospectively studied the outcome of these patients in a resource constrained setting.METHODSThe case records of all newly diagnosed multiple myeloma patients who fulfilled the criteria for active myeloma between January 2018 and December 2020 were identified. The diagnosis of double hit and triple hit myeloma was based on mSMART classification (Presence of two or more of the following: IgH-FGFR3 translocation, IgH-MAF translocation, TP53 deletion, gain of chromosome 1q). Their case records were retrieved and information regarding baseline characteristics, therapy and outcomes was noted.RESULTSA total of 55 patients with newly diagnosed DH/THM were treated at our center during the study period. Median age of the cohort was 60 years with almost an equal number of male and female patients (M= 26; F= 29). Renal failure (serum creatinine >2.0mg/dl) was present in 33 patients (60%) while bone lesions, anemia (Hemoglobin <10 gm/dl) and hypercalcemia (serum calcium >12mg/dl) were present in 44 (80%), 51 (92.7%) and 24 (43.6%) patients respectively. Six patients (10.9%) fulfilled the criteria for plasma cell leukemia.All but 2 patients had gain of 1q with at least 3 copies. The most common cytogenetic combination was IgH-FGFR3 translocation with gain of 1q, which was seen in 28 patients (50.9%). This was followed by co-occurrence of TP53 deletion with gain of chromosome 1q in 11 patients (20%). Nine patients (16.4%) had triple hit myeloma.Bortezomib was used in the initial therapy of 43 patients (78.2%) and IMiDs (lenalidomide, thalidomide and pomalidomide) were used in the initial therapy of 32 patients (58.2%). Most patients received triplet therapy (N=32; 58.2%) with the most common regimen being RVd. Nine patients (16.4%) died within the first month of diagnosis and another 7 died in next month [15 patients in 2 months (29.1%)]. The most common cause of death within 2 months was progressive disease (N=13; 81.3%). Twenty-two patients (40%) achieved VGPR or better with anti-myeloma therapy and 7 patients (12.7%) underwent autologous HCT. Of the 39 evaluable patients, 21 patients (53.8%) relapsed during follow up with a median EFS of 8 months. Of the 7 patients who underwent transplant, 5 patients have had a follow-up of more than 1 year, of whom 3 have relapsed. One patient with post-transplant relapse died with progressive disease and CMV colitis.Median follow up of the entire cohort was 11 months (Range- 0 to 35 months). After excluding patients who died within the first 2 months of diagnosis, the median follow up was 14 months (Range- 3 to 35 months). Thirty-three patients (60%) patients died during follow-up. The most common cause of death was active or progressive disease (25 patients, 78.1%). Median OS for the cohort was 13 months. On univariate analysis, survival was better for patients without renal injury at presentation (29 months vs 6 months; p=0.007) and patients with <5% circulating plasma cells (14 months vs 2 months; p=0.045). Patients who achieved VGPR had a better OS than patients who did not (Not reached vs 3 months; p=0.000) as did patients who underwent auto transplant (Not reached vs 10 months; p=0.023). OS did not significantly differ with TP53 deletion status, or number of copies of 1q (3 or greater than 3). In the multi-variate analysis, presence of renal failure (Hazard ratio- 2.663; p-0.017) and >5% circulating plasma cells (Hazard ratio- 3.082; p-0.020) were significantly associated with increased risk of mortality, while achievement of VGPR or better with therapy was associated with longer survival (Hazard ratio- 0.174; p-0.001).DISCUSSIONThe outcome of DH/THM remains poor in the real-world setting. The outcome was not affected by specific high-risk cytogenetic abnormalities or their combination in our study. Progressive disease within the first two months of diagnosis was the most common cause of death in more than 1/3 rd of the patients. Novel therapies and protocols are required to improve outcomes for this group of multiple myeloma patients. DisclosuresNo relevant conflicts of interest to declare.

OAB-055: Gain and amplification of 1q induce transcriptome deregulation and worsen the outcome of newly diagnosed Multiple Myeloma patients

OAB-055: Gain and amplification of 1q induce transcriptome deregulation and worsen the outcome of newly diagnosed Multiple Myeloma patients

Advanced Forms of the Myeloproliferative Neoplasms Are Associated with Chromosomal Abnormalities Involving 1q and 12q: Implicating MDM2 and MDM4 in Disease Progression

Inevitably patients (pts) with myeloproliferative neoplasms (MPNs) including polycythemia vera (PV) and essential thrombocythemia (ET) progress to more advanced forms of myelofibrosis (MF) and acute myeloid leukemia (AML). Loss of TP53 function plays a critical role in cancer biology yet, the MPNs are characterized predominately by wild type (WT) TP53 (Rampal et al, PNAS, 2014;111: 5401-10). The major protein regulators of TP53 are MDM2 and MDM 4. TP53 activity has been reported to be down-regulated in MPNs due to overexpression of MDM2 (Lu et al, Blood, 2012;120:3098-3105, Nakatake et al, Oncogene, 2012: 311: 323-33). Since the gene for MDM4 is located on chromosome 1q and the gene for MDM2 is located on chromosome 12q, we hypothesized that cytogenetic abnormalities involving these locations might lead to disease progression This hypothesis is supported by the recent observation that mice engineered to possess JAK2 V617F and lack TP53 are prone to develop AML as compared to JAK2 V617F+ mice with WT TP53 (Rampal et al, PNAS, 2014:111:5401-10). Gain of 1q (+1q) in the form of jumping translocations in pts with MPN was previously reported to be associated with transformation to AML (Najfeld et al, Br.J. Haematol., 2010; 151: 285-291).These studies were extended to 362 MPN pts enrolled in 5 different MPN-RC clinical trials involving pts with ET or PV (222), MF and MPN -Blast Phase (MPN-BP)(140) and 989 pts with MPNs, ET(177), PV (400), MF (385) and MPN-BP(27) who were studied at the Icahn School of Medicine at Mount Sinai (ISMMS) between 1986 and June 2017. 92 pts in the various MPN-RC trials (25.4%) and 344 (34.7%) of the pts from ISMMS were chromosomally abnormal. Among the abnormalities, 23 pts in the MPN-RC trials had either +1q/dup(1q)(n=16) or rearrangements of 12q (n=7) (6.3% of the total pts and 25.0% of the chromosomally abnormal pts). Similarly, the most frequently observed chromosomal abnormality at ISMMS associated with disease progression, either as a sole abnormality or as a subclone with other chromosomal abnormalities, was gain of 1q, identified in 59 pts (6.0% of total, 17.4% of the abnormal pts). Gain of 1q was observed in three forms: 1) as an unbalanced translocation with the breakpoint at 1q21. 2) a duplication of 1q region and/or 3) as a jumping +1q translocation. The most frequent unbalanced translocation identified was +der(9)t(1;9)(q21;q12) (24%) resulting in three copies of 1q and 3 copies of 9p. Rearrangement of 12q most frequently involved balanced translocations and/or inv(12q) with 50% of these pts having a breakpoint at 12q15 where the gene for MDM2 is located. Gain of 1q/dup(1q) and rearrangements of 12q were mutually exclusive with the exception of one patient with a translocation between chromosomes 1 and 12 [der(12)t(1;12)(q24;q15)]. Among PV/ET pts enrolled in MPN-RC trials, 1.3% (3/222) had +1q and 0.0% (0/222) had 12q abnormalities, while 11% (15/140) of pts enrolled in MF and MPN-BP treatment protocols had +1q and 5% (7/140) had 12q abnormalities. These findings indicate that +1q and structural abnormalities of 12q are associated with more advanced forms of MF and MPN-BP (p<0.001). In both, the MPN-RC and the ISMMS data bases, 42 cases with +1q and 13 cases with 12q abnormalities were identified where the driver mutational status was known. The JAK2 V617F mutation was observed in 83.3% (35/42)of pts with +1q while 23.1% (3/13) pts with 12q abnormalities were JAK2 V617F+.These data indicate that gain of 1q but not 12q abnormalities are associated with the JAK2 V617F mutation (p<0.001). In a limited number of pts (n=10) we were able to monitor the chronological acquisition of 1q and 12q abnormalities. The 1q abnormalities occurred 5-7 years following the initial cytogenetic evaluation of PV pts and the proportion of cells with gain of 1q increased over time being present in 100% of cells over the next 4-5 years of observation. By contrast, 12q abnormalities occurred exclusively in MF pts as the sole abnormality and appeared in 90-100% of the examined cells at presentation. In 5 MF pts with +1q, qPCR demonstrated upregulation of MDM 4 mRNA transcripts (86%) when compared to 5 pts without +1q. These data suggest that MPN disease progression is associated with specific chromosomal abnormalities that result in up regulation of MDM2 or MDM4 which further down-regulate p53 .These data provide the rationale for the use of MDM2/MDM4 antagonists as a means of halting MPN disease progression. DisclosuresMascarenhas:CTI Biopharma: Research Funding; Novartis: Other: DSMB member , Research Funding; Merck: Research Funding; Incyte: Other: Clinical Trial Steering Committee , Research Funding; Promedior: Research Funding; Janssen: Research Funding.

Impact of Gain and Amplification of 1q in Newly Diagnosed Multiple Myeloma Patients Receiving Carfilzomib-Based Treatment in the Forte Trial

Impact of Gain and Amplification of 1q in Newly Diagnosed Multiple Myeloma Patients Receiving Carfilzomib-Based Treatment in the Forte Trial

Gain of Chromosome 1q is associated with early progression in multiple myeloma patients treated with lenalidomide, bortezomib, and dexamethasone

Gain of chromosome 1q (+1q) is commonly identified in multiple myeloma and has been associated with inferior outcomes. However, the prognostic implication of +1q has not been evaluated in the setting of standard triplet regimens. We retrospectively analyzed 201 consecutive patients with newly diagnosed myeloma who received induction with lenalidomide, bortezomib, and dexamethasone (RVD) and were tested for +1q at diagnosis by fluorescent in-situ hybridization. Patients with +1q (n = 94), compared to those without +1q (n = 107), had shorter median progression-free survival (PFS) (41.9 months vs 65.1 months, p = 0.002, HR = 1.90) and overall survival (median not reached (NR) for either arm, p = 0.003, HR 2.69). In subgroup analyses, patients with co-occurring +1q and t(4;14), t(14;16) or del(17p) or with 4 or more copies of 1q had significantly worse PFS (25.1 months and 34.6 months, p < 0.001 and p = 0.0063, respectively), whereas patients with three copies and no other high-risk cytogenetic abnormalities had no significant difference in PFS. These data suggest that when treated with RVD induction, patients with +1q should be considered at very high risk for early progression in multiple myeloma when ≥4 copies are detected or in the context of other high-risk cytogenetic abnormalities.

An Acquired High-Risk Chromosome Instability Phenotype in Multiple Myeloma: Jumping 1q Syndrome

IntroductionChromosome instability (CIN) is a driver of copy number aberrations (CNAs) in cancer, and is a major factor leading to tumor heterogeneity and resistance to therapy. By definition, CIN is an increased rate or ongoing acquisition and accumulation of CNAs and not simply the existence of structurally and numerically abnormal aneuploid clones. In multiple myeloma (MM), the most common whole-chromosome CNAs involve either hyperdiploid or non-hyperdiploid clones. Secondary segmental CNAs are associated with high-risk (HR) in MM and involve gains of 1q21 and deletions of 17p (del17p). These types of intra-chromosomal segmental CNAs are also found in the CIN phenotypes of the autosomal recessive (AR) chromosome instability syndromes. These syndromes include Fanconi anemia, Bloom's syndrome, and ICF syndrome (Immunodeficiency, Centromeric instability and Facial anomalies). These chromosome instability syndromes display a spectrum of aberrations characterized by higher rates of chromosomal breaks, chromatid exchanges, quadriradials, and pericentromeric aberrations. In particular, patients with ICF syndrome show a marked increase of 1q12 pericentromeric instability including 1q12 decondensation, triradials, multibranched chromosomes 1q, and 1q micronuclei. ICF patients also show transient 1q aberrations including isochromosome 1q (iso1q) and unbalanced translocations of 1q to 9q and 16q. In MM, we have previously reported increasing pericentromeric instability during tumor progression resulting in increasing CNAs of 1q21 by unbalanced jumping translocations of 1q12 (JT1q12). Strikingly, in a subset of MM patients with 1q21 CNAs of ≥ 5 a distinct cytogenetic phenotype emerges which demonstrates transient 1q12 aberrations including 1q12 decondensation, triradials, and multibranched chromosomes 1q morphologically identical to those seen in ICF patients. In MM this chromosome instability leads to a cascade of increasing clonal 1q21 duplications, iso 1qs, and unbalanced 1q translocations with 16q and 17p, resulting in losses in these receptor chromosomes (RC) and massive intra-clonal CNA heterogeneity.MethodsTo investigate the cytogenetic impact and progression of high CNAs of 1q21, we performed a comprehensive metaphase analysis of 50 patients showing segmental aneuploidies with 4 or more copies of 1q by G-banding. Locus specific FISH and spectral karyotyping were used to identify the key transient unstable and clonal structural aberrations of 1q12 resulting in segmental aneuploidies in the derivative RCs. Probe for 1q12 (Vysis) was used according to the manufacturer's protocol. Locus specific BAC clones for 1q21 (CKS1B) and 17p (TP53) were prepared and analyzed as previously described (Sawyer et al., Blood 123: 2014). IGH translocations were investigated with IGH break apart probes (Vysis).ResultsData for 50 patients including CNAs of 1q21 of ≥ 4, IGH translocations, del(17p), derivative RCs, are presented. The t(4;14) was found in 15 patients, del(17p) in 23, and both aberrations were found in 8 patients. All patients showed unbalanced gains of 1q and deletions of RCs, the most frequent being 7 patients with der(1;16) and 6 with iso1q. In four of the 23 patients with del(17p), the deletion was due to a JT1q12 to 17p. Seven patients with 1q21 CNAs of ≥ 5 showed profound instability involving the 1q12 satellite DNA, demonstrating both transient and clonal aberrations driving the 1q21 CNAs. These aberrations included unstable 1q21 triplications, JT1q12s, iso1q formation with intra-arm 1q12 CNAs, and region specific breakage-fusion-bridge cycle amplifications.ConclusionsAmong patients with ≥ 5 CNAs of 1q21, a subset develop an acquired HR chromosome instability phenotype with an elevated rate of 1q12 pericentromeric instability characterized by concomitant deletions in 16q, iso1q, del(17p), and intra-arm segmental instability. These patients show pronounced instability in the 1q12 satellite DNA, morphologically identical to ICF syndrome, suggesting hypomethylation of this region as a driver of both 1q21 CNAs and deletions in RCs. We hypothesize that region specific hypomethylation of 1q12 provides the genomic background for the onset of an acquired 1q12 chromosome instability phenotype in MM similar to that found in ICF syndrome. For myeloma patients demonstrating this 1q12 chromosome instability phenotype we propose the term “jumping 1q syndrome.” DisclosuresEpstein:University of Arkansas for Medical Sciences: Employment. Davies:Amgen: Consultancy, Membership on an entity's Board of Directors or advisory committees; ASH: Honoraria; Abbvie: Consultancy; TRM Oncology: Honoraria; Celgene: Consultancy, Honoraria, Membership on an entity's Board of Directors or advisory committees; Takeda: Consultancy, Membership on an entity's Board of Directors or advisory committees; MMRF: Honoraria; Janssen: Consultancy, Honoraria. Morgan:Takeda: Consultancy, Honoraria; Celgene: Consultancy, Honoraria, Research Funding; Janssen: Research Funding; Bristol-Myers Squibb: Consultancy, Honoraria.

Hyperhaploid Multiple Myeloma (MM): A Rare Karyotypic Subgroup Retaining Disomy 18 and 1q12∼23 Amplification

Abstract 3983In MM, chromosome ploidy levels are commonly used in the cytogenetic risk stratification of the disease. Two distinct ploidy groups occur: a hyperdiploid group, associated with a better prognosis, and a hypodiploid group, associated with a poor prognosis. The hyperdiploid group (47–57 chromosomes) is characterized by a consistent set of odd-numbered chromosomes including trisomies for chromosomes 3,5,7,9,11,15,19, and 21. This group is also characterized by fewer structural aberrations and is found in 50% to 60% of patients with metaphase aberrations. The hypodiploid group (35–45 chromosomes) encompasses clones composed of either hypodiploid, pseudodiploid, and/or near-tetraploid variants. The hypodiploid group has more frequent structural chromosome aberrations involving adverse IGH translocations and deletions of 17p. We have identified a group of 22 patients by routine G-banding with hyperhaploid karyotypes that exhibit a range of 30–34 chromosomes with a modal number of 32. The hyperhaploid clones are characterized by the same set of odd-numbered chromosomes found in hyperdiploid MM, including 3, 5, 7, 9, 11, 15, 19, and 21, however all these chromosomes are found in disomy instead of trisomy. The single notable exception is the retention of chromosome 18 in the hyperhaploid karyotypes. In seventeen of the 22 patients, both a hyperdiploid clone and a hyperhaploid clone were identified in the same sample, with the hyperdiploid clone always representing the dominant cell line. Five patients showed only a hyperhaploid clone. Importantly, both the hyperdiploid and hyperhaploid clones in these patients shared the same set of structural aberrations. This suggests the hyperhaploid karyotypes originated from the hyperdiploid clones by the loss of a single normal copy of each chromosome pair. Chromosome 18 was retained in 18 of 22 patients, and all or part of 1q was retained or newly amplified in five patients. Ten of 22 patients also exhibited the loss or deletion of chromosome 5, which indicates this is an additional secondary event in the hyperhaploid clones. Additionally, we investigated the hyperhaploid clones utilizing fluorescence in-situ hybridization (FISH) and spectral karyotyping in the nine samples that adequate sample was available. FISH probes for IGH rearrangements indicated that only two of the nine cases had IGH translocations, one with a t(4;14), while in the other case a receptor of the IGH signal was not identified. Deletions of 17p (TP53) were found in all nine samples. FISH for 1q21 (CKS1B) was informative in five cases and showed a 1q copy number (CN) of 2 in three cases, of 3 in one case, and of 2–6 in one case. The only recurring structural aberrations identified in the hyperhaploid clones were aberrations of 1q. Segmental disomy for 1q12∼23 was retained in the sole chromosome 1 in two patients (cryptic to G-banding), and whole-arm jumping 1q (JT1q12) was identified in 2 cases. One particularly informative case with a JT1q21 demonstrated a CN of 2–6 for 1q21 and instability of the 1q12 pericentromeric heterochromatin in the form of triradials of 1q. The multi-radials of 1q were the origin of multiple extra acentric copies of 1q and acentric isochromosomes 1q. The findings in this study indicate that a progression of chromosome aberrations exists in hyperhaploid MM just as it does in hyperdiploid and hypodiploid MM. The hyperhaploid clones apparently originate from a single catastrophic event in which an entire haploid set of chromosomes is lost from a hyperdiploid clone, with the striking exception of the retention of chromosome 18. In patients with structural chromosome aberrations, the same set of aberrations is retained in the hyperhaploid cells, including 1q12∼23 amplification. In a subset of patients, these clones may then undergo a loss or deletion of chromosome 5. Finally, hyperhaploid clones can also undergo genomic instability in the form of JT1q12 translocations involving whole-arm CN increases of 1q. The retention of disomy for only chromosome 18 and 1q12∼23 amplification suggests these two specific chromosome aberrations may be important in the survival of these clones. In this group of 22 patients, the median survival time from diagnosis was 32 months, suggesting a poor prognosis for hyperhaploid MM. Disclosures:No relevant conflicts of interest to declare.

Multiple copies of duplicated 1q in myelodysplastic syndrome

Multiple copies of duplicated 1q in myelodysplastic syndrome

Jumping Translocations 1q12 Contribute to Copy Number (CN) Alterations in Multiple Myeloma (MM): Unexpected Focal Amplifications of Receptor Chromosomes (RC)

Abstract 298MM is characterized by complex chromosome 1 aberrations including an unbalanced rearrangement known as a jumping translocation 1q12 (JT1q12). The JT1q12 is a chromosomal mechanism for arm-length amplification of 1q by the duplication of copies of 1q onto one or more non-homologous chromosomes. Two types of JT1q12 usually occur: one results in the translocation of the entire 1q to the telomere of a RC, increasing the CN of genes on the entire 1q. The second type of JT1q12 results from a whole-arm unbalanced (arm-length) translocation which increases the CN of genes on 1q, while at the same time decreasing the CN of genes on the RC. An increase in CN of 1q21 in MM has been reported to be associated with disease progression and poor prognosis. An alternative mechanism for CN alterations of 1q21 is also known that includes the focal amplification of a 1q12∼23 amplicon, which includes the genes MCL1, BCL9, IL6R, MUC1, PSMD4, and CKS1B, among others. In fact, focal amplifications of 1q21 and other genomic regions have recently been found to be shared across multiple cancer types, suggesting common amplification targets in the cancer genome (Nature, 463: 899–905, 2010). Unfortunately, little is known about the mechanisms involved in focal amplifications of 1q21 or other amplifications known to occur during tumor progression. We therefore undertook a comprehensive metaphase analysis to study the cytogenetic progression of the JT1q12 aberrations. We investigated 70 patient specimens showing 1q aberrations by G-banding. Metaphase studies utilizing fluorescence in-situ hybridization (FISH) and spectral karyotyping (SKY) techniques revealed duplications of RCs which were cryptic to G-banding. Sixty-nine cases showed 1q21 amplification of CKS1B with a CN of 1q21 ranging from 3–12 per metaphase cell. Twenty patients showed a 1q21 CN of 3, 19 patients showed a CN of 4, 16 patients with a CN of 5, 5 patients with a CN of 6, 6 patients with a CN of 7–8, 2 patients with a CN of 9–11, and 1 patient with a CN of 12. Thirty-five patients showed JT1q12 related deletions in RCs,11 patients with 16q-, 6 patients lost 19q-, 3 patients lost 6q-, and 2 patients each with 5q- and 8p-. Five patients showed focal amplifications of the 1q12∼23 amplicon by a breakage-fusion-bridge (BFB) cycle mechanism. The resulting focal amplifications were inverted duplications of extended ladder-like structures with 2–8 copies of the 1q12∼23 amplicon. These cases showed site specific breakage in the 1q12 pericentromeric heterochromatin, which mediated the organization of the amplicon by bracketing both the proximal and distal ends of the amplicon. One case of particular interest demonstrated BFB cycles incorporating alternating segments of chromosome 16q11 and the 1q12∼23 amplicon, demonstrating the simultaneous amplification of segments of two non-homologous chromosomes. The amplified segments of both chromosomes 1q12∼23 and 16 then translocated together to multiple RCs, resulting in arm-length deletions in RCs. The unexpected novel finding in this study was the juxtaposition of JT1q12 pericentromeric heterochromatin with the distal segment of a RC resulting in the focal amplification of the adjacent RC segment. In 4 patients the distal duplicated segment of the RC and the adjacent JT1q12 subsequently translocated to new RCs, thus resulting in the simultaneous duplication of chromosome segments both proximal and distal to the 1q12 pericentromeric heterochromatin. Three of these focal duplications involved the distal 8q, including the locus of MYC, and one involved the distal 18q, including the locus of BCL2. This newly identified type of focal amplification provides evidence that 1q12 pericentromeric instability can also provide a mechanism for the amplification of chromosome segments on RCs and thereby simultaneously increase the CN of genes thought to play a role in the progression of MM, including MYC, BCL2, and the 1q12∼23 amplicon. Disclosures:Shaughnessy:Myeloma Health, Celgene, Genzyme, Novartis: Consultancy, Employment, Equity Ownership, Honoraria, Patents & Royalties. Barlogie:Celgene: Consultancy, Honoraria, Research Funding; IMF: Consultancy, Honoraria; MMRF: Consultancy; Millennium: Consultancy, Honoraria, Research Funding; Genzyme: Consultancy; Novartis: Research Funding; NCI: Research Funding; Johnson & Johnson: Research Funding; Centocor: Research Funding; Onyx: Research Funding; Icon: Research Funding.

Jumping translocations of the long arms of chromosome 1 in myeloid malignancies is associated with a high risk of transformation to acute myeloid leukaemia*

Jumping translocations of the long arms of chromosome 1 in myeloid malignancies is associated with a high risk of transformation to acute myeloid leukaemia*

Whole Arm Duplication of 1q in Myeloid Neoplasm, with Emphasis On Derivative (1;7)(q10;p10).

Abstract 4238 BackgroundDuplication of the long arm of chromosome 1 (1q) is a recurrent finding in hematologic neoplasms. It is most commonly observed as an unbalanced whole arm translocation with chromosome 7, resulting in concurrent deletion of 7q (ISCN as der(1;7)(q10q10)). It is less frequently observed as an unbalanced translocation with a chromosome other than chromosome 7 or as a duplication of only a segment of 1q. The der(1;7) has been described in myelodysplastic syndrome (MDS), and acute myeloid leukemia (AML), and is reportedly less commonly associated with myeloproliferative neoplasms (MPN). The effect of this rearrangement on prognosis is unclear, although it is classified by the International Scoring System (IPSS) as a karyotypic variant of del(7q)/-7 and is thus assigned poor risk. We have initiated a thorough review of all cases of duplication of the long arm of chromosome 1 seen at our institution. MethodsAfter approval from the Institutional Review Board, all cases seen at Mayo Clinic over a 20 year period (1989-2009) were searched for a duplication of the long arm of chromosome 1. The sole exclusion criterion was a complex karyotype (3 or more independent chromosome abnormalities in the clone). Cytogenetic and hematopathology review was done and relevant clinical data were abstracted from the Mayo Clinic record. Survival of MDS patients and MPN patients was determined from time of cytogenetic diagnosis of der(1;7) until death or last contact with the patient, using the Kaplan-Meier method. ResultsAmong 23,375 unique patients at Mayo Clinic with a cytogenetic study for hematologic neoplasm, 229 (0.98%) patients carried a duplication of 1q. Within this group, 99 (0.42%) had duplication of part of 1q, 123 (0.53%) carried a whole arm duplication of 1q, and 7 patients carried an isochromosome 1q (resulting in 4 copies of 1q). The most common karyotype among those with a whole arm duplication of 1q was der(1;7) (n=84), followed by similar whole arm translocations involving chromosomes 9 and 19 (n=7 each) or chromosomes 12, 13, 14, 15, 16, 17, 18, 20, 21, or 22 (n=1 to 6 each). We focused on 65 der(1;7) patients who had adequate hematopathological material that could be reviewed and sub-classified according to WHO criteria. MDS was identified in 30 patients and MPN in 20. MDS (therapy related, RARS, RAEB-1 or 2) was seen in 9 patients (14%), MDS/ refractory cytopenia with multilineage dysplasia (RCMD) in 19 (29%). MDS/Myeloproliferative neoplasm (MPN) in 2 (3%), MPN not otherwise classified in 5 (8%), and MPN / primary myelofibrosis (including 1 post-polycythemic and one post-thromobocythemic) (PMF) in 15 (23%). The bone marrow was apparently normal in 5 patients (8%), CLL was noted in 3 (5%), and 1 each exhibited features of systemic mast cell disease or myeloma. Acute myeloid leukemia (AML) was identified 5 patients (8%). The M:F ratio of the der(1;7) patients was 2.8:1. The median age at occurrence of the der(1;7) was 63 years (range: 13-87 years). All 5 AML patients have died (median survival 9 months). Of the MDS and MPN patients, the median survival was 2.2 and 2.8 years, respectively, from the time of cytogenetic diagnosis. Overall survival was not significantly different between the MDS and MPN group, and they exhibited 30% survival after 5 years (see Figure). ConclusionsDuplication of 1q was observed in nearly 1% of the Mayo Clinic patients who had a bone marrow cytogenetic study for a hematologic neoplasm. Among those with a der(1;7), AML was uncommon but MDS and MPN were similar in frequency. Median survival of 2.2 to 2.8 years for the MDS and MPN group justifies assignment to a more favorable group than the current IPSS assignment. [Display omitted] Disclosures:No relevant conflicts of interest to declare.

CD32B on Clonal Plasma Cells in Systemic Light-Chain Amyloidosis (AL) and Multiple Myeloma (MM): A Target for Immunotherapy in Both Disorders and a Prognostic Marker in MM.

Despite stem cell transplant and new therapies, nearly all patients with AL and MM die of disease or complications of treatment. Novel approaches that selectively kill clonal plasma cells are needed. CD32B, the inhibitory Fcγ receptor IIB, is a member of the Fc receptor (FcR) family on chromosome 1q. B-cells and some monocytes and dendritic cell subtypes express cell-surface CD32B, which has a cytoplasmic inhibitory motif important in regulating immune responses. Unlike the CD32B2 isoform on myeloid cells, CD32B1 on B-cells is not internalized, making it a suitable target for monoclonal antibody (MAb) therapy (Blood 2006Jun6 Epub). CD32B has not previously been found on AL and MM plasma cells. We used purified CD138+ marrow and blood cells from AL, MM and plasma cell leukemia (PCL) patients, and 5 human MM cell lines, to evaluate CD32B gene and cell-surface expression with gene expression profiles (GEP) (Affymetrix U133 PLUS 2.0), RT-PCR for CD32B1 and B2, and flow-cytometry with the 2B6 MAb for CD32B. In AL, GEP showed that CD32B expression was significantly higher than other FcR genes (p<0.01), RT-PCR showed that CD32B1 was prominently expressed on CD138+ cells from both newly diagnosed and relapsed AL patients, and flow cytometry showed intense cell-surface staining for CD32B on >98% of all AL plasma cells. In MM, public GEP data (http://lambertlab.uams) showed that CD32B expression was significantly higher than other FcR genes while RT-PCR showed CD32B1 message in CD138+ MM and PCL specimens but not in the RPMI 8226 cell line. Cytogenetic analysis then showed that 8226 cells lack t(4;14) but have 4 copies of 1q, implicating segmental uniparental tetrasomy of a mutant allele in the lack of CD32B message. Flow cytometry showed median 96% CD32B expression on CD138+ marrow cells from patients with normal or hyperdiploid cytogenetics, but significantly lower expression (median 69%, p=0.01) in patients with unfavorable cytogenetics (del 13, t(4;14)). CD32B expression was lower still on PCL specimens (median 10%) and nil on MM cell lines. We then investigated the GEP of sorted CD138+/CD32B+ and CD32B− fractions from MM patients. In CD138+/CD32B+ cells, CD45C and CXCR4 were overexpressed. In CD138+/CD32B− cells, genes on chromosome 1q were overexpressed, including those for cancer testis antigens and others possibly associated with biologic aggressivity (Cancer Cell 2006; 9:313). Notably, the CD32B-specific MAb 2B6 directs human mononuclear cell cytotoxicity against CD32B+ cell lines, reduces tumor growth and improves tumor-free survival in a mouse xenograft model. Moreover, an Fc-engineered humanized variant of 2B6 elicits ADCC-mediated specific cytotoxicity against a low-level CD32B+ MM cell line in vitro. In sum, these data show that CD32B is important in AL and MM; that CD32B on AL and the majority of MM plasma cells provides a target for MAb therapy; and that, given the results in MM, PCL and MM cell lines, CD32B expression is inversely related to biologic aggressivity. Genetic instability of chromosome 1q may provide a direct basis for this relationship thereby mechanistically linking CD32B expression in MM to prognosis. These data support further evaluation of CD32B and clinical studies of anti-CD32B MAb therapy in AL and MM.

Atypical extraventricular neurocytoma with oligodendroglioma-like spread and an unusual pattern of chromosome 1p and 19q loss

An insular cortex tumor in a 54-year-old woman showed unequivocal neurocytic features, including open nuclei, distinct nucleoli, and strong synaptophysin immunoreactivity. Ultrastructurally, there were neuritic-type processes with microtubules and hillock-like attachments, and there were dense-core granules. Atypical features were mitotic activity, prominent vasculature, and small foci of necrosis. Peripherally, there was oligodendroglia-like histology with single-cell infiltration of white matter and perineuronal spread in cortex. Fluorescence in situ hybridization analysis with chromosome 1 and 19 probes showed 3 copies of 1q and 2 copies of 1p and showed 2 copies of 19q and 4 copies of 19p. This yielded a 1p-19q loss of heterozygosity pattern similar to that seen in oligodendrogliomas, although the actual chromosomal abnormality is distinct. This tumor, best classified as an atypical neurocytoma with oligodendroglia-like spread, supports suggestions of a close histogenic relationship between oligodendroglial and neurocytic tumors. This case also illustrates the limitations of relying exclusively on loss of heterozygosity analysis for tumor classification.