Metaphase Karyotype Research Articles

Somatic chromosome counts and karyomorphological analysis of 12 Allium (Amaryllidaceae) taxa

The genus Allium is a well-known model system for cytological research, and its cytological characteristics are considered important for species identification. In this study, we counted chromosome numbers of 21 populations from 12 taxa comprising seven sections (Butomissa, Sacculiferum, Schoenoprasum, Oreiprason, Reticulatobulbosa, Rhizirideum, Tenuissima) and five subgenera (Butomissa, Cepa, Polyprason, Reticulatobulbosa, Rhizirideum) from Kyrgyzstan, Mongolia, and South Korea. The chromosome numbers varied with 2n=16 or 2n=32. New ploidy levels were found in A.karelinii (2n=2x=16) and A.burjaticum (2n=2x=16) from Kyrgyzstan and Mongolia, respectively. Photomicrophotographs of metaphase chromosomes and karyotype analyses of all Allium taxa studied are provided.

A treatment-refractory aggressive MDS-MLD with multiple highly complex chromosome 5 intrachromosomal rearrangements: a case report

BackgroundA patient with a myelodysplastic neoplasm exhibited a karyotype with multiple complex chromosome 5 rearrangements. This patient appeared to have a catastrophic cytogenetic event that manifested as a treatment-refractory aggressive form of disease, which lead to patient demise within one year. Both the clinical presentation and disease course were unusual based on the medical history and morphologic findings. Such cases of myelodysplastic syndrome with multilineage dysplasia (MDS-MLD) with complex abnormalities are not reported in the literature.Case presentationThe patient was a 62-year-old female who presented with pancytopenia and dyspnea. The morphologic appearance of the peripheral blood smear and bone marrow biopsy, along with flow cytometric findings, favored the diagnosis of MDS-MLD unclassifiable. Myelodysplastic syndrome (MDS) with multilineage dysplasia (MDS-MLD), is an MDS characterized by one or more cytopenias and dysplastic changes in two or more of the myeloid lineages (i.e., erythroid, granulocytic, and megakaryocytic). The bone marrow, in particular, showed prominent dysplasia, including the presence of atypical megakaryocytes with small hypolobated morphology reminiscent of those typically seen in MDS with isolated 5q deletion. Cytogenetic analysis, including interphase and metaphase FISH, karyotype and SNP chromosomal microarray were performed, as well as DNA sequencing studies. Cytogenetic analysis showed a very complex karyotype featuring multiple 5q intrachromosomal rearrangements including a pericentric inversion with multiple interspersed deletions and monosomy 7. FISH studies showed a partial deletion of the PDGFRβ gene, and SNP chromosomal microarray and targeted panel-based sequencing identified biallelic loss of function of the TP53 gene. Based on the pathologic findings, the patient was treated for MDS but did not respond to either lenalidomide or azacitidine.ConclusionThe genetic changes described, in particular, the complex intrachromosomal rearrangements of chromosome 5, suggest the occurrence of a sudden catastrophic event that led to an aggressive course in the patient’s disease. Conventional karyotyping, metaphase and interphase FISH, SNP chromosomal microarray and NGS helped to identify the complex genetic changes seen in this case. This highlights the importance of utilizing a multimodality approach to fully characterize complex chromosomal events that may significantly impact disease progression, treatment and survival.

Low Intensity Salvage Therapy after Failure of Standard (7 +3) Induction Regimen in Acute Myeloid Leukemia (AML). a Single Center Experience from United Arab Emirates

Hypomethylating agent (HMA) and venetoclaxR as salvage therapy for Acute Myeloid Leukemia (AML) failing intensive induction therapy. A single center experience from United Arab Emirates. Introduction: Patients with AML who are younger and fit for therapy are treated with intensive chemotherapy. Complete remissions (CRs) range from 40-80% (Kantarjian, Blood vol. 116,22 (2010): 4422-9). Non-CR patients can be treated with salvage therapy as a bridge to Allogeneic hematopoietic cell transplant (AlloHCT). Salvage regimens have included high dose cytarabine based therapies (FLAG, MEC, HAM) which can be more toxic limiting the use of AlloHCT. Lower intensity and more targeted therapy could decrease toxicity with improved responses. We describe our experience with a low intensity regimen as salvage treatment after failure of intensive induction therapy and bridge to AlloHCT. Methods: This is a retrospective analysis of AML patients treated at Tawam Hospital, UAE from 01/2019 to 06/2022. Patient data was collected from the medical records including: demographics, hemogram at presentation, bone marrow biopsies, karyotyping, treatment and response. Results: Eighty patients were diagnosed with AML during this period. The median age was 45 years (range 16-95 years). Fifty-seven patients received therapy while twenty-three patients refused or were not eligible due to comorbidities and / or performance status. Intensive chemotherapy (infusional cytarabine and idarubicin 7+3 or FLAG) was used for thirty-three patients. The median age was 35 years (range 16 - 63 years). Cytogenetics (Metaphase Karyotyping and FISH): normal, n=7; good risk n=6 (inv 16 or t (8;21), growth failure n=5, poor risk n=10 (Complex = 3, Del 7 = 3, 11q abnormalities = 3, trans 6;9 =1) other n=5. Molecular testing for NPMI, FLT3, IDH1 and IDH2 was not available. Three patients died during induction and are excluded from analysis. Fifteen patients attained CR after 1 cycle of induction (50%). Twelve patients not in CR received outpatient, low intensity salvage therapy with HMA (5 azacitidine 75 mg/m2 daily x 7 days subcutaneously with Venetoclax 100 mg orally daily x 21 days along with antimicrobial prophylaxis with azoles in 28 days cycles). Bone marrow was repeated after the second cycle. The median number of cycles is 3 (range 1-9). Eight (67 %) achieved CR1. There were no treatment related deaths. This is comparable to our report of FLAG salvage therapy (Hassan IB. Int J Hematol. 2018 Oct;108(4):390-401). Four of the eight patients who achieved remission went for AlloHCT while one is awaiting AlloHCT. One patient is unable to go for HCT and is in remission after 9 cycles of therapy. Two patients after achieving CR relapsed and died of disease progression. With a median follow up of 123 days (range 33 -1107 days) five patients are alive and in remission, one has been lost to follow-up while the other six have died from progressive disease and complications. Conclusion. Low Intensity therapy with HMA and Venetoclax is a reasonable and effective alternative to intensive chemotherapy as first line salvage with minimal morbidity and zero treatment related mortality. Treatment was well tolerated and no patients required hospitalization for complications. This regimen is easily administered as an outpatient, serving as a bridge to AlloHCT

Accurate Detection of Clinically Actionable Copy Number Variants in Diverse Hematological Neoplasms By Routine Targeted Sequencing: A Comparative Performance Study

Background: Accurate genotyping of Copy Number Variations (CNVs) is an essential part of the clinical evaluation of diverse hematologic malignancies. Conventional karyotyping and FISH are current gold standards to assess CNVs, but both have some limitations. Next-generation targeted sequencing panels are commonly incorporated into current clinical practice in most cancer centers. We examined the utility of CANARy (Copy number ANomaly Assessment and Recovery), a CNV profiling method for three hematological malignancies using targeted panel sequencing data. Patients and Methods: We studied patients with myelodysplastic syndrome (MDS), chronic lymphocytic leukemia/small lymphocytic lymphoma (CLL/SLL), and plasma cell disorders (PCD) identified from a cohort of 4752 hematological specimens profiled at Stanford University by conventional cytogenetics and next-generation sequencing (NGS) between 2018 and 2021. We curated a cohort of 200 patients diagnosed with MDS (n=72), CLL (n=46), and PCDs including MM, AL amyloidosis, and MGUS (PCD, n=82), along with 50 cytogenetically normal controls. Specimens were bone marrow aspirates, peripheral blood, or FFPE tissue. All samples were characterized by conventional metaphase karyotyping and/or FISH and we focused on the analysis of clinically relevant CNV (-5q, -7/-7q, -20q, +8 for MDS; -11q22, -13q14, -17p13, +12 for CLL; and +1q21, -1p32, -13q14, -17p.13 for PCD). The specimens separately had clinical NGS (164 genes, Heme-STAMP) performed no more than four weeks from standard cytogenetic analysis. We applied multiple CNV enumeration tools to sequencing data for CNV genotyping at these loci, including our recently developed method CANARy (Chabon et al, Nature 2020). CANARy is a read depth-based, genome-wide CNV caller from targeted sequencing data that combines information from on-target and off-target fragments to assign a final Z-score and copy number index to a set of pre-defined genomic regions. We benchmarked CANARy alongside three others publicly available CNV callers: CNVkit (Talevich et al, PLoS Comput Biol. 2016), ichorCNA (Adalsteinsson et al, Nat Commun. 2017), and WISECONDOR (Straver et al, Nucleic Acids Res. 2014). All tools were run using the default parameters and following author recommendations. We calculated the accuracy and other statistics parameters for each CNV caller, considering karyotype/FISH as the true reference. We estimated tumor fraction using two strategies: the variant allele fraction (VAF) of disease-related driver mutations and using the ichorCNA estimate. Results: A total of 211 CNVs were identified by karyotype and/or FISH, with disease-specific distribution summarized in Fig. A. CANARy detected 148/211 (70.1%), CNVkit 88/211 (41.7%), ichorCNA 116/211 (55%) and WISECONDOR 158/211 (74.9%). The overall accuracy for CANARy, CNVkit, ichorCNA and WISECONDOR was 86.3%, 81.8%, 84.3% and 83.1% respectively. CANARy had both the highest overall accuracy (86.3%) and overall F1-score (0.73) (Fig.B). WISECONDOR had the highest number of false positives (83 vs. CANARy = 48). Interestingly, all CNV tools performed better in MDS specimens, with 92% accuracy for CANARy in MDS samples, likely related to the higher neoplastic purity of these samples. The median ichorCNA tumor fraction estimates for MDS, CLL and PCD were 22% (0-99%), 14% (0-70%), and 7% (0-94%), respectively. In addition, CNV calling was inferior in PCD tumors than other tumor types. Within CLL, -11q22 and -13q14 lesions were more challenging to detect with any of the four methods, with CANARy sensitivity of 50% and 31.3% respectively, likely related to the smaller minimal deletion regions of these lesions. The 17p deletion in CLL cases, known for its poor prognostic impact, was best identified by CANARy; with a 100% sensitivity and a 97.6% specificity. The only non-concordant result for -17p was a false positive that was detected by all 4 NGS CNV genotypers but not FISH, possibly suggesting a false negative FISH result. Conclusions: CANARy accurately identifies clinically significant CNVs in common hematologic neoplasms and could thus serve as a complementary approach to the current cytogenetic evaluation. CANARy uses data already generated from targeted sequencing panels and therefore does not impose additional work or cost. Low tumor fraction could be a limitation for CNV calling. Figure 1View largeDownload PPTFigure 1View largeDownload PPT Close modal

Astragalus stenosemioides (Fabaceae)’in karyotip analizi

In this research the metaphase chromosome number and karyotype of Astragalus stenosemioides species growing naturally in Turkey are studied. The study revealed that the chromosome number of A. stenosemioides is 2n = 16. The basic chromosome number of species is determined as x = 8. The karyotype formula of A. stenosemioides is 8m. The total length of the somatic chromosomes of A. stenosemioides ranges between 1.96 - 3.38 μm. The total haploid chromosome length of A. stenosemioides is determined as 21.36 μm. In addition, the karyotype asymmetry of the species was evaluated by different methods; Stebbins classification, TF%, AsK%, Syi and Rec, A1, A2, DI, A, and AI.

The importance of cytogenetic and molecular aberrations in multiple myeloma

Multiple myeloma (MM) is a heterogeneous clonal malignancy of plasma cells characterized by cytogenetic and molecular abnormalities. Chromosomal abnormalities are present at diagnosis and can evolve during the progression of MM. Metaphase karyotyping and fluorescence in situ hybridization are considered the standard diagnostic procedures performed in clinical practice. These test results are required to determine the Revised International Staging System classification, treatment algorithms, and short- and long-term prognoses. Given the dynamic development of cytogenetic and molecular research, we should expect further progress in better understanding the biology of MM and changes to patient care in the coming years.

Abstract 3006: Melanoma cell lines derived from patient derived xenografts (PDX) can contain murine chromosomes resultant from cell fusions

Abstract Introduction: Patient Derived Xenografts (PDX) provide a distinct platform to generate in vivo and ex vivo cell lines to investigate novel therapies. However, human-murine cell fusions may skew the models and is a poorly characterized phenomenon. Methods: Malignant melanoma IRB-consented biopsies [IRBs: 17-000548; 17-001685; 17-001351; 11-003254] were excised from various sites and implanted into gender-matched 4-6 weeks old NOD-scid IL2Rgammanull mice. Tumor-bearing mice were humanely euthanized and adherent cell lines were generated after Miltenyi gentleMACS™ dissociation of the PDXs. Briefly, adherent cells are allowed to proliferate and passaged when 70% confluent for approximately 4-8 weeks. PDX-derived cell products were processed for long term-cryostorage and an aliquot was sent for GenePrint 10 System-Promega®-LaragenTM cell line authentication paired with patient-matched peripheral blood mononuclear cells (PBMC) for specimen congruency. Mouse-genome contaminant resulting cultures were prepared and sent for cell line characterization by G-banded metaphase karyotyping-Cell Line GeneticsTM. Cultures containing murine karyotypes were processed according to manufacturer recommendations by Miltenyi Mouse Cell Depletion KitTM where the flow through (negative fraction) represents the human-cell fraction. Results: We implanted 86 patient-derived biopsies into mice over 3years, from which 39 [45%] resulted in progressively growing PDXs. From these PDXs we have generated 26 [37%]-patient-specific PDX-derived cell lines. G-banded metaphase karyotyping demonstrated that 6 [24%]- cell lines presented murine chromosome contamination. The immunoprofile of these 6 cellular materials appear to be murine as the totality of the cultures were sequestered in the column by mouse depletion cell cocktail. Separation assay was validated by bonafide human cell lines, utilized as human positive controls, M257 and MX210 both with human karyotypes and matched Cell ID profiling. The murine colon carcinoma cell line MC38, was utilized as a positive murine control, and performed identically as the 6 PDX-derived cellular materials with abnormal mouse genomes. Conclusion: Detection of murine-human fusion cells in cell lines derived from PDXs may limit the utility of these models. Citation Format: Agustin Vega-Crespo, Ignacio Baselga-Carretero, Ivan Perez-Garcilaso, Sameeha Jilani, Kevin Chen, Vaneet Khullar, Egmidio Medina, Yaroslav Teper, Antoni Ribas, Begoña Comin-Anduix. Melanoma cell lines derived from patient derived xenografts (PDX) can contain murine chromosomes resultant from cell fusions [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2021; 2021 Apr 10-15 and May 17-21. Philadelphia (PA): AACR; Cancer Res 2021;81(13_Suppl):Abstract nr 3006.

Interim PET-CT Result Is Not Predictive of Survival in Patients with MYC-Rearranged Non-Burkitt Aggressive B Cell Lymphoma

Abstract Background Patients with a diagnosis of MYC-rearranged non–Burkitt aggressive B-cell lymphoma (MYC-R), including those with double hit lymphoma, are at high risk of developing relapsed/refractory disease, even if treated with intensive front-line immunochemotherapy. It is common in clinical practice and clinical trials to perform an interim positron emission tomography (PET)/computed tomography (CT) scan (iPET) during front-line therapy for diffuse large B-cell lymphoma. However, the utility of the iPET result for MYC-R patients for predicting outcomes is unclear. Patients and Methods We performed a single-center retrospective study with centralized pathologic review and PET/CT image acquisition and interpretation for 28 MYC-R patients. The patients received front-line therapy with R-CHOP (rituximab, cyclophosphamide, hydroxydaunorubicin [doxorubicin], Oncovin [vincristine], prednisone) or intensive immunochemotherapy. Results Eight patients had iPET-positive (iPET+) and 20 patients had iPET-negative (iPET−) results using the Deauville visual assessment criteria. At a median follow-up length of 30.4 months, progression-free survival was 65% and overall survival was 76%, neither of which differed significantly between the iPET− and iPET+ patients. The positive predictive value of iPET for progression at 30 months was 25%, and the negative predictive value was 65%. Conclusion Although patients with MYC-R lymphoma have been reported to be at high risk of primary treatment failure, this was not predicted by iPET+ results. Thus, the iPET result should not be used to guide changes in front-line or consolidative therapy for these patients.

The Impact of Cytogenetic Clonality on Outcomes in Younger Acute Myeloid Leukemia Patients: Results on 1290 Patients Included in the EORTC/Gimema AML-10 and AML-12 Trials

Background. A recent large retrospective study has observed that the presence of cytogenetic clonal heterogeneity, as detected by metaphase karyotyping, was associated with poor prognosis in patients (pts) with AML (Bochtler et al., J Clin Oncol 2013, 31: 3898-905).Objectives. The aims of the current study were to reassess the impact of cytogenetic clonal heterogeneity on outcomes in younger AML pts (age 15-60 yrs) and to determine how clonal heterogeneity modified the effects of the type of remission-induction chemotherapy and of having a donor in an independent sample of AML pts < 60 years of age with an abnormal karyotype.Methods. In the AML-10 trial pts were randomized to receive either daunorubicin (DNR), mitoxantrone (MTX), or idarubicin (IDA) in addition to standard-dose cytarabine and etoposide for induction chemotherapy. In the AML-12 trial, pts were randomized between SDAC and high dose cytarabine (HiDAC) induction, both with DNR and etoposide. In both trials, a second cycle of the same induction chemotherapy was administered in patients who achieved a PR. Patients who achieved a CR or a CRi after 1 or 2 courses of induction chemotherapy received a consolidation chemotherapy with the same anthracycline as in the induction course plus intermediate dose cytarabine. Thereafter, patients were scheduled to receive in first CR/CRi an allogeneic HCT if they had an HLA-identical donor or an autologous HCT otherwise. Cytogenetic analyses were performed at diagnosis and centrally reviewed. For the current analysis, cytogenetics were re-classified using the MRC classification. The definition of a (sub)clone was according to the international System of Human Cytogenomic Nomenclature (2016). Abnormal karyotypes were classified as no subclone (cytogenetic abnormality present in a single clone), subclones (presence of multiple (2, 3 or 4) clones) and composite karyotypes (CK) when clonal heterogeneity was too complex to allow enumeration of individual subclones.Results. In the AML-10 trial, 2157 pts were randomized to receive DNR, MTX or IDA. The current analysis was performed in a subgroup of 684 pts with an abnormal karyotype: 555 of them had no subclones, 109 at least 1 subclone and 20 a CK. In the AML-12 trial, 1942 pts were randomized between HiDAC and SDAC. The current analysis was performed in a subgroup of 606 pts with an abnormal karyotype: 470 of them had no subclones, 117 at least 1 subclone and 19 a CK (Table 1). Pts with a CK (n=39) were older than 25 years and 84.6% had adverse MRC cytogenetic features. In comparison to pts with no subclones, those with subclones had a similar probability of achieving a CR and had similar DFS from CR/CRi and OS from randomization (Table 1). However, in comparison to pts with no subclones, those with a CK had a similar probability of achieving a CR/CRi but a shorter DFS (HR=2.10, 95% CI: 1.39-3.16) and OS (HR= 1.54, 95% CI: 1.07-2.21). In multivariate Cox model stratified by protocol and adjusted for age, cytogenetic risk group, secondary versus primary AML and PS, neither the presence of subclones (HR 1.11, 95% CI: 0.92-1.33) nor CK (HR 0.93, 95% CI: 0.64-1.34) were associated with shorter OS (P=0.49). Comparable findings were obtained after restricting the analyses to pts with adverse/intermediate cytogenetic features.Among pts included in the AML-10 trial, the impact on the OS of the type of anthracycline used (IDA vs MTZ vs DNR) was not significant, and its magnitude was not statistically different according to the clonal heterogeneity status (no subclones versus subclones versus CK)(P=0.24). Further, among pts included in the AML-12 trial, the difference regarding OS between HiDAC and SDAC was limited (HR=0.94), and its magnitude was not impacted by the clonal heterogeneity status (P=0.54). Finally, in the whole cohort, DFS from CR/CRi was significantly longer in the subgroup of pts with a donor (n=250) than in those without a donor (n=449) (HR=0.72, 95% CI: 0.58-0.89, P=0.002); the impact on DFS of having a donor vs no donor was similar in the 3 clonal subgroups (P-value for interaction from the Cox model: P=0.26).Conclusions. The influence of clonal heterogeneity on outcome was evaluated in an independent sample of AML patients. Clonal heterogeneity as defined by the presence of subclones showed no effect on OS as compared to cytogenetic abnormal patients without clonal subclones. The dismal outcome in the pts with a CK could be explained by the known predictors of poor prognosis. [Display omitted] DisclosuresNo relevant conflicts of interest to declare.

Fluorescence in Situ Hybridization (FISH) Frequently Upstages Risk Estimates for MDS Patients with Normal Metaphase Karyotype

IntroductionThe international prognostic scoring system (IPSS) and its revised version (R-IPSS) are the most widely used clinical models for MDS risk stratification. Both models account for cytogenetic abnormalities captured by G-banding metaphase karyotyping. Among MDS patients, 50-60% will have a normal karyotype. MDS FISH panels are commonly applied to supplement cytogenetics. In small subsets of patients with normal karyotype, MDS FISH may detect cytogenetic abnormalities not resolved by metaphase karyotype. The utility of MDS FISH panel in refining IPSS or R-IPSS cytogenetic risk score among those patients is not known. We explored the impact of FISH in upstaging MDS patients with a normal karyotype and its prognostic value if incorporated into IPSS or R-IPSS cytogenetic risk score.MethodsWe identified patients with normal karyotype by G-banding who had cytogenetic abnormalities detected by an MDS FISH panel. The MDS FISH panel included probes for deletion (del(5)), del(7), del (20), trisomy 8 and del 17p. IPSS and R-IPSS was calculated based on karyotype as the standard recommendation and then recalculated according to cytogenetic abnormalities detected by the MDS FISH panel (FISH_IPSS and FISH_R-IPSS). Median overall survival (OS) was calculated from time of diagnosis using Kaplan-Meier estimates.ResultsAmong 1600 patients in our MDS database with normal karyotype by G-banding, 53 patients (3%) had a cytogenetic abnormality detected by FISH. The cytogenetic abnormalities detected included del(7)/monosomy 7 in 25/53 patients (47%), del(5q)/monosomy 5 in 22/53 patients (42%), trisomy 8 in 14/53 (26%), del(20q) in 10/52 (19%), and 17p deletion in 1/10 (10%).According to IPSS, 15 patients (33%) were classified as low risk, 18 patients (40%) as intermediate-1 (int-1), 9 patients (20%) int-2, and 3 patients (7%) as high risk. Table-1 summarizes restaging using FISH_IPSS where 53% of low risk IPSS were upstaged to int-1, 56% of int-1 upstaged to int-2, and 78% of int-2 upstaged to high risk.By conventional R-IPSS, 18 patients (40%) were very low risk, 11 patients (24%) low risk, 12 patients (26%) intermediate risk, and 4 patients (9%) high risk. Upon restaging using FISH_R-IPSS (Table 2), 61% of very low risk patients, all low risk patients, 92% of intermediate risk patients, and 50% of high risk patients with FISH abnormalities were upstaged.Among patients with low risk IPSS, median OS was 64 months (mo) for those low risk by FISH_IPSS and 61 mo for those upstaged to int-1 by FISH_IPSS. For patients with int-1 risk IPSS, median OS was 26 month for those int-1 by FISH_IPSS and 24.8 mo for those upstaged to int-2 risk by FISH_IPSS. Finally, for patients with int-2 risk IPSS, median OS was 30 mo for those classified as int-2 and 22 mo for those upstaged by FISH_IPSS.Based on R-IPSS the median OS was 64, 24, 25 and 19 mo for very low, low, int, and high risk groups. Using FISH_RIPSS the median OS was 64, 61,30,21,22 mo respectively for very low, low, int, high and very high risk groups (p.003). Based on R-IPSS cytogenetic risk category all those patient were classified as good risk given normal karyotype with median OS of 30 mo. Based on FISH data if cytogenetic score was re-calculated accordingly, 16 patients remained good risk, 22 patients intermediate risk, and 14 patients high risk. The median OS was 30, 30 and 22 mo respectively.ConclusionsFISH detected cytogenetic abnormalities in a small number of patients with normal karyotype by G-banding, which altered risk score assignment by IPSS and R-IPSS in a significant subset of positive patients. [Display omitted] DisclosuresKomrokji:Celgene: Honoraria; Novartis: Honoraria, Speakers Bureau. Sallman:Celgene: Research Funding. Padron:Incyte: Honoraria, Research Funding. Sweet:Ariad: Consultancy, Speakers Bureau; Karyopharm: Consultancy, Research Funding; Otsuka: Consultancy; Incyte: Research Funding; Pfizer: Consultancy; Novartis Pharmaceuticals: Consultancy, Speakers Bureau. Lancet:Pfizer: Other: Institutional research funding, Research Funding; Boehringer Ingelheim: Consultancy; Novartis: Consultancy; Celgene: Consultancy; Erytech: Consultancy; BioSight: Consultancy; Bio-Path Holdings: Consultancy; Janssen: Consultancy; Jazz Pharmaceuticals: Consultancy.

Lenalidomide, Bortezomib, and Dexamethasone (RVD) As Induction Therapy in Newly Diagnosed Multiple Myeloma (MM)

Background: Lenalidomide, bortezomib and dexamethasone (RVD) was demonstrated to be an effective and well tolerated induction regimen in both transplant eligible and ineligible patients. Two large randomized phase III trials show an overall response rate (ORR) >95% (Durie et al, Attal et al) supporting the combination. We have evaluated our institutional data of patients treated with RVD induction therapy.Methods: 751 newly diagnosed MM patients were treated with RVD induction therapy [R - 25 mg/day (days 1-14), V - 1.3 mg/m2 (days 1, 4 8, 11) and D - 40 mg once/twice weekly as tolerated every 21 days] from January 2008 until April 2015. Dose-adjustments were made based on the treating physicians discretion and patient tolerability. Demographic and outcomes data for the patients were obtained from our IRB approved myeloma database and responses were evaluated per IMWG Uniform Response Criteria.Results: The median age was 61 years (range 22-83). Other patient characteristics include: M/F 54%/46%; W/AA 58%/30%; ISS I/II/III 36%/24%/19%; Isotype IgG/IgA/FLC 56%/21%/17%; high risk/standard risk 28%/62%. Among the standard risk patients, t(11;14):13%, del (13): 30% comprised majority of the patients. Among high risk patients, the distribution of high risk cytogenetic abnormalities are t(4;14)/t(14;16)/del(17p): 5%/2.5%/10.5%. Complex karyoptye by conventional metaphase karyotyping was seen in 16% of the patients. A total of 562 patients (74.8%) underwent ASCT1 and 165 patients (22%) were offered deferred transplant. Among the patients that opted for deferred transplant, 56 patients (33.9%) received ASCT1 at first relapse. Median time to receive ASCT1 among these relapsed patients was 30 months (3-96 months). For others, median time for patients to receive ASCT1 was 5 months (range, 1-64 months). Post-ASCT, 70% patients were on maintenance therapy tailored to their risk (R-54%; RVD-8%; V-4%). This differed from the deferred ASCT patient population (mostly standard risk patients), where 66% of the patients received revlimid based maintenance. Median PFS1 among the deferred group was 73 months (range, 54.3-91.62). At a median follow up of 44 months for the entire cohort, the estimated median PFS was 56 months (49.85-62.15 months). Median PFS for standard risk patients was 68 months (60.62-75.37) versus 33 months for the high-risk patients (28-37.9 months), p<0.001 and median OS has not been reached, but encouraging results were seen for both high-risk and standard risk cohort with 64% and 77% of patients alive at the 84-month mark, respectively (Fig 1). Response rates are summarized in table 1.Conclusions: This is the largest reported cohort of myeloma patients treated with RVD regimen and highlights the activity of this induction regimen with superior response rates of close to 90% ≥VGPR rates post-ASCT. This regimen should serve as an important bench mark for future studies designing optimal induction regimens. [Display omitted] DisclosuresHeffner:ADC Therapeutics: Research Funding. Boise:Abbvie: Consultancy; Eli Lilly and Company: Research Funding. Kaufman:Amgen, Novartis: Research Funding; Amgen, Roche, BMS, Seattle Genetics, Sutro Biopharma, Pharmacyclics: Consultancy. Nooka:Amgen, Novartis, Spectrum, Adaptive tecnologies: Consultancy.

Outcomes of Myelodysplastic Syndromes (MDS) Patients with Chromosome 17 Abnormalities: A Report on Behalf of the MDS Clinical Research Consortium (MDSCRC)

IntroductionChromosome (ch) 17 cytogenetic abnormalities (CA) are reported associated with poor outcomes in MDS patients. The Revised International Prognostic Scoring System (R-IPSS) classifies ch 17 CA in the intermediate cytogenetic risk group; however, the majority of patients harbor the ch 17 CA in the context of a complex karyotype, which is considered high risk in the R-IPSS. The TP53 gene is located on the short arm of ch 17, and somatic TP53 mutations or deletions are associated with poor outcomes in MDS, including poor survival after allogeneic stem cell transplant. Recently, a high response rate was described in patients with TP53 mutations and acute myeloid leukemia treated with decitabine (Welch J et al NEJM 2016). Response rates to hypomethylating agents (HMA) are not well characterized among MDS patients with ch 17 CA. We report the outcome and prognosis of ch 17 CA in a large, multi-institutional cohort of MDS patients.MethodsWe identified patients with ch 17 abn within the MDSCRC database and compared them to those without ch 17 abn. CA were identified by G-banging metaphase karyotype analysis. Baseline characteristics were collected at time of diagnosis or prior to any therapy. Response rates to HMA were assessed using International Working Group criteria (IWG 2006), where overall response (OR) was defined as complete remission (CR), marrow CR, partial remission or hematological improvement (HI). Chi -square and t-test were used for comparing baseline characteristics, Kaplan-Meier estimates were used for calculating overall survival (OS) from time of diagnosis and cox regression was used for multivariable analysis.Results:We identified 352 MDS patients with ch 17 CA and compared them to 2053 patients with all other CA. Table-1 summarizes baseline characteristics between the two groups. As expected, most MDS patients with ch 17 CA were classified as higher risk R-IPSS. Among those who had molecular testing, 95 patients with ch 17 CA (81%) had TP53 somatic mutations compared to 143 patients (19%) with other CA. (p <.001). The rate of AML transformation was 36% among ch 17 CA group compared to 19% among those without ch 17 CA (p<.001)The ch 17 CA abnormality was most frequently observed as part of a complex karyotype; 303 patients (86%) had complex karyotype (>3 CA). The ch 17 CA observed included monosomy 17 in 147 patients (42%), add(17p) in 84 patients (24%), trisomy 17 in 29 patients (8%), i(17) in 10 patients (3%), while 91 patients (26%) had other ch 17 CA including chromosomal translocations. Among patients with monosomy 17, 143/147 patients (97%) had complex karyotype.188 MDS patients with ch 17 CA were treated with HMAs and were compared to 597 patients with other CA treated with HMAs. The OR was 45% and 41%, respectively (p =.42). CR rates were higher among MDS patients with ch 17 CA: 26% compared to 18% for those with other CA (p=.025). The median OS for patients with ch 17 CA was 12 mo (95% CI 11-14) compared to 25 mo (95% CI 23-29) in patients with other CA.Among 227 MDS patients with detailed ch 17 CA data, the median OS was 12 mo for patients with complex karyotype (n=204) compared to 26 mo for those with ch 17 CA non-complex karyotype (n=23), p=.001. There was no difference in median OS between patients with monosomy 17 and add17p.In Cox regression analyses adjusting for R-IPSS, Ch 17 CA presence was associated with inferior overall survival, HR 1.5 (95% CI 1.3-1.8), p <.005. The main driver of poor outcome was the presence of TP53 mutations.In a cox regression analysis accounting for R-IPSS, ch 17 CA and TP 53 mutation, The HR of death was 1.7 (p=.003) for TP 53 mutation, and for Ch 17 CA HR 1.1 (p=.7)ConclusionsIn MDS patients, ch 17 CA are often observed in the setting of complex karyotype and are associated with poor outcomes. The majority of patients with ch 17 CA harbor TP53 mutations, which drive poor outcome. CR rates to HMAs were higher among patients with ch 17 CA, but median OS was inferior compared to those with other CAs, similar to TP53 mutated AML patients. [Display omitted] DisclosuresKomrokji:Novartis: Honoraria, Speakers Bureau; Celgene: Honoraria. Steensma:Incyte: Equity Ownership; Janssen: Consultancy, Research Funding; Onconova: Consultancy; Amgen: Consultancy, Membership on an entity's Board of Directors or advisory committees; Celgene: Consultancy; Pfizer: Consultancy, Membership on an entity's Board of Directors or advisory committees; Takeda: Consultancy; H3 Biosciences: Consultancy; Pfizer: Consultancy; Novartis: Consultancy, Membership on an entity's Board of Directors or advisory committees. Roboz:Cellectis: Research Funding; AbbVie, Agios, Amgen, Amphivena, Array Biopharma Inc., Astex, AstraZeneca, Celator, Celgene, Clovis Oncology, CTI BioPharma, Genoptix, Immune Pharmaceuticals, Janssen Pharmaceuticals, Juno, MedImmune, MEI Pharma, Novartis, Onconova, Pfizer, Roche Pharmace: Consultancy. Sallman:Celgene: Research Funding. Padron:Incyte: Honoraria, Research Funding. Sekeres:Celgene: Membership on an entity's Board of Directors or advisory committees.

Fusion gene detection by RNA-sequencing complements diagnostics of acute myeloid leukemia and identifies recurring NRIP1-MIR99AHG rearrangements.

Identification of fusion genes in clinical routine is mostly based on cytogenetics and targeted molecular genetics, such as metaphase karyotyping, fluorescence in situ hybridization and reverse-transcriptase polymerase chain reaction. However, sequencing technologies are becoming more important in clinical routine as processing time and costs per sample decrease. To evaluate the performance of fusion gene detection by RNAsequencing compared to standard diagnostic techniques, we analyzed 806 RNA-sequencing samples from patients with acute myeloid leukemia using two state-of-the-art software tools, namely Arriba and FusionCatcher. RNA-sequencing detected 90% of fusion events that were reported by routine with high evidence, while samples in which RNA-sequencing failed to detect fusion genes had overall lower and inhomogeneous sequence coverage. Based on properties of known and unknown fusion events, we developed a workflow with integrated filtering strategies for the identification of robust fusion gene candidates by RNA-sequencing. Thereby, we detected known recurrent fusion events in 26 cases that were not reported by routine and found discrepancies in evidence for known fusion events between routine and RNA-sequencing in three cases. Moreover, we identified 157 fusion genes as novel robust candidates and comparison to entries from ChimerDB or Mitelman Database showed novel recurrence of fusion genes in 14 cases. Finally, we detected the novel recurrent fusion gene NRIP1- MIR99AHG resulting from inv(21)(q11.2;q21.1) in nine patients (1.1%) and LTN1-MX1 resulting from inv(21)(q21.3;q22.3) in two patients (0.25%). We demonstrated that NRIP1-MIR99AHG results in overexpression of the 3' region of MIR99AHG and the disruption of the tricistronic miRNA cluster miR-99a/let-7c/miR-125b-2. Interestingly, upregulation of MIR99AHG and deregulation of the miRNA cluster, residing in the MIR99AHG locus, are known mechanisms of leukemogenesis in acute megakaryoblastic leukemia. Our findings demonstrate that RNA-sequencing has a strong potential to improve the systematic detection of fusion genes in clinical applications and provides a valuable tool for fusion discovery.

Blast MRD AML-1 Trial: Blockade of PD-1 Added to Standard Therapy to Target Measurable Residual Disease in Acute Myeloid Leukemia (AML) 1- an Investigator-Initiated, CTEP-Sponsored, Randomized Phase 2 Study of the Anti-PD-1 Antibody Pembrolizumab in Combination with Conventional Intensive Chemotherapy (IC) As Frontline Therapy in Patients with Acute Myeloid Leukemia (AML)

Study Background: The presence of minimal residual disease (MRD) after IC in AML patients is often a harbinger of relapse, and therapeutic targeting of MRD has emerged as an important strategy to improve long-term outcomes. The importance of the PD-1/PD-L1 pathway in immune evasion of AML has been demonstrated in murine AML models whereby PD-L1 and PD-1 were upregulated in leukemia cells, and PD-1 blockade suppresses invivo leukemia cell proliferation and improved survival in AML bearing mice (Zhang et al., 2009; Zhou et al., 2010). Combining IC with immune checkpoint inhibition (ICPI) has resulted in a significant clinical activity in some advanced solid malignancies including non-small-cell lung cancer. A recent single-arm phase 2 study combining IC with nivolumab showed the combination was feasible and resulted in undetectable MRD in 53% of patients with Complete response (CR)/CR with incomplete count recovery (CRi) (Ravandi et al, 2019). Here we report on the design of the first randomized multi-center clinical trial of IC+ICPI in fit AML patients. Methods: The primary objective of this investigator-initiated Cancer Therapy Evaluation Program (CTEP)-sponsored multi-institutional, randomized phase II study (NCT04214249) is to assess the percentage of patients with MRD negative CR (MRD- CR) as measured by flow cytometry at the end of first cycle of consolidation therapy with IC+ pembrolizumab and compare between the two study arms (Figure 1). Secondary objectives include rates of CR/CRi, complete remission with partial recovery count (CRh) and hematologic improvement (HI) to red blood cells and platelets, MRD negativity at Day 14, MRD-negative CR at end of induction therapy and MRD negative CR after last consolidation cycle, event free survival (EFS), relapse free survival (RFS), duration of response (DOR), and overall survival (OS), and proportion of patients who develop severe toxicity. Important exploratory objectives include MRD assessment by duplex sequencing (DS) and comparing DS and multiparameter flow cytometry for MRD detection. The study will consist of an induction, consolidation, and maintenance phase of therapy. A total of 124 patients will be included (62 patients in the intervention arm and 62 patients in the control arm). Planned stratification factors include 1) age (younger than 65 vs. 65 and older), 2) cytogenetics by FISH or metaphase karyotype (presence vs. absence of core binding factor inversions and translocations), and 3) t-AML or AML arising from prior/antecedent MDS. All patients will receive cytarabine at 100 mg/m2/day continuous infusion on Days 1-7 and either idarubicin 12 mg/m2/day IV on Days 1-3 or daunorubicin 60 mg/m2/day IV on Days 1-3. on Day 8 of the induction chemotherapy, half of the patients will receive pembrolizumab IV (intervention arm), and continue Q3 weeks throughout Induction/Consolidation, and thereafter for up to 2 years from the start of maintenance. The other half will not receive pembrolizumab (control arm). After 31 efficacy-evaluable patients (patients who had a bone marrow biopsy done after the first consolidation cycle) in each of the two study arms have had their primary endpoint results available, a formal futility and toxicity analysis will be conducted. Follow up will continue up to 2 years after the last patient is randomized. Eligible patients are aged ≥18 and ≤75 years with newly diagnosed and pathologically-confirmed AML, including secondary AML arising from prior MDS and therapy-related AML. Patients with FLT3-mutated AML are excluded as well as those with prior allogeneic stem cell transplant, active infection requiring systemic therapy, and requiring use of high dose corticosteroids. Responses in AML patients will be assessed using European Leukemia Net 2017 response criteria. EFS will be calculated as the time from initial treatment to either disease relapse or death. OS will be calculated from time from initial treatment to death. Biomarker analyses will include an extensive array of immunologic and correlative studies that will evaluate PD-1 and PD-L1 expression, immune cell subset analysis, leukemia specific T-cell response, cytokine level, T-cell receptor (TCR) repertoire, as well as RNA-seq, gut microbiome characterization and metabolomics, and whole exome sequencing for tumor and germline DNA. Figure 1 Disclosures Zeidan: Epizyme: Consultancy, Honoraria; Astellas: Consultancy, Honoraria; Daiichi Sankyo: Consultancy, Honoraria; Cardinal Health: Consultancy, Honoraria; Acceleron: Consultancy, Honoraria; Leukemia and Lymphoma Society: Other; Jazz: Consultancy, Honoraria; Celgene / BMS: Consultancy, Honoraria, Research Funding; Abbvie: Consultancy, Honoraria, Research Funding; Otsuka: Consultancy, Honoraria; Pfizer: Consultancy, Honoraria, Research Funding; Seattle Genetics: Consultancy, Honoraria; BeyondSpring: Consultancy, Honoraria; Cardiff Oncology: Consultancy, Honoraria, Other; Takeda: Consultancy, Honoraria, Research Funding; Ionis: Consultancy, Honoraria; MedImmune/Astrazeneca: Research Funding; Trovagene: Consultancy, Honoraria, Research Funding; Aprea: Research Funding; ADC Therapeutics: Research Funding; Astex: Research Funding; Incyte: Consultancy, Honoraria, Research Funding; Taiho: Consultancy, Honoraria; CCITLA: Other; Novartis: Consultancy, Honoraria, Research Funding; Boehringer-Ingelheim: Consultancy, Honoraria, Research Funding; Agios: Consultancy, Honoraria. Radich:Amgen: Consultancy; Bristol-Myers Squibb: Consultancy; Jazz: Consultancy; Novartis Pharmaceuticals Corporation: Consultancy, Research Funding.

Is There an Unequal Benefit of Autologous Stem Cell Transplant in Different Cytogenetic Groups of High Risk Patients with Multiple Myeloma: The University of Miami Experience

BACKGROUND: Stratification using cytogenetics (CG), either metaphase karyotyping or fluorescence in situ hybridization (FISH) is used to identify patients (pts) with multiple myeloma (MM) who are at higher risk of relapse and tend to have poorer survival. It is largely unknown if autologous stem cell transplantation (auto-HCT) after high dose melphalan (200 mg/m2) is able to modify the survival of some of these high-risk MM pts and to make it comparable to standard risk. METHODS: Pts were classified as high risk (HR) if either conventional cytogenetics or FISH demonstrated at least one of the following 1q+, 1p-, 17p-, 13q-, t(4;14), or t(14;16), realizing that the inclusion of 13q- by FISH alone and 1p- in the HR MM definition is controversial. Pts with normal chromosomes or those with trisomies and hyperdiploidy were considered standard-risk (SR). We compared progression-free survival (PFS) and overall survival (OS) via a retrospective analysis of pts at the University of Miami Sylvester Comprehensive Cancer Center who underwent auto-HCT between January 2014 and December 2017 for MM. Survival analyses were performed using the log-rank test, with significance at p-value &amp;lt; 0.05. RESULTS: Male pts comprised 56% of the population, and 40% of pts were of Hispanic ethnicity. Of 205 pts undergoing auto-HCT, 108 (53%) had at least one HR cytogenetic abnormality. Interestingly, the depth of response to pre-transplant induction was higher in pts classified as HR, with 71% (77 of 108) achieving at least a very good partial response (VGPR), while 24% (23 of 96) SR pts achieved VGPR. While OS remained largely unaffected in HR pts (34.0 m vs. 35.1 m, p = 0.27); HR pts had an inferior PFS compared to SR pts (21.9 m vs. 25.7 m, p = 0.041). The presence of trisomies did not negate the poorer PFS of HR pts. When we evaluated specific HR CG abnormalities, OS and PFS in patients with 1q+ or t(4;14) were surprisingly comparable to SR pts, indicating a significant benefit from auto-HCT. On the other hand, OS was significantly decreased in pts with 1p- when compared to standard risk (16.5 m vs. 35.1 m, p = 0.004) or other high-risk patients (16.5 m vs. 35.4 m, p = 0.01), implying that 1p- group derive no benefit from auto-HCT. Similarly, OS was shorter in pts with t(14;16) (16.5 m vs. 34.4 m, p = 0.025) and with 17p- (26.6 m vs. 35.1 m, p=0.01), however the PFS was not affected in these populations. In pts with 13q-, PFS was significantly shorter (20.3 m vs. 25.7 m, p=0.023) compared to SR pts without affecting OS. CONCLUSION: At our center in a retrospective analysis of 205 pts: a) patients with HR MM responded better and faster than SR pts to induction, b) those with 1p- did not derive any benefit from transplant and c) pts with 17p- and t(14;16) had some short term benefit (similar PFS to SR group) but at the end their OS remained inferior. Nevertheless, we consider a very important finding the fact that, by having auto-HCT, pts with 1q+ and t(4;14), equalized their PFS and OS to those of SR pts. Based on these findings, patients with 1q+ and t(4;14) should still have a transplant in CR1 even after optimal induction. Whether 17p- and t(14;16) pts can extend PFS benefit after transplant into OS benefit with maintenance regimens stronger than lenalidomide alone remains to be determined. Disclosures Hoffman: Celgene: Honoraria, Speakers Bureau; Loxo: Current equity holder in publicly-traded company; Seattle Genetics: Research Funding.

Fluorescence in Situ Hybridization (FISH) Utility for Risk Score Assessment in Patients With MDS With Normal Metaphase Karyotype

BackgroundCytogenetic profile is an essential parameter in myelodysplastic syndromes (MDS) risk stratification by both International Prognostic Symptom Score (IPSS) and Revised (R)-IPSS. Almost one-half of patients with MDS have normal cytogenetics by metaphase karyotype. Here we report the yield of MDS fluorescence in situ hybridization (FISH) panel detecting cytogenetic abnormalities in these patients and its impact on risk stratification. Patients and MethodsAmong patients with normal metaphase karyotype, we assessed those patients who had cytogenetic abnormalities detected by an MDS FISH panel, which included probes for del (5), del (7), del (20), trisomy 8, and del (17p). Risk stratification was calculated by both IPSS and R-IPSS. ResultsOf 1600 patients with MDS with normal metaphase karyotype, 53 (3%) patients had cytogenetic abnormality detected by MDS FISH panel. Integrating the MDS FISH panel cytogenetics (IPSS + FISH restaging) resulted in upstaging the score, where 53% of low-risk IPSS were upstaged to intermediate (int)-1, 56% of int-1 were upstaged to int-2, and 78% of int-2 were upstaged to high risk. Based on the R-IPSS, 61% of very low-risk patients, all low-risk patients, 92% of intermediate-risk patients, and 50% of high-risk patients with FISH abnormalities were upstaged, respectively. ConclusionThe yield of MDS FISH panel detecting cytogenetic abnormalities in patients with normal karyotype by G-banding is low and may not warrant ordering the panel in all patients. Among the 3% of patients with normal karyotype who had cytogenetic abnormality detected by FISH, the risk score assignment by IPSS and R-IPSS was upstaged.

Actividad bactericida del agua de plata coloidal sobre indicadores de calidad microbiológica: un estudio a partir de muestras ambientales

El objetivo del presente trabajo fue evaluar la actividad bactericida del agua de plata coloidal sobre los microorganismos indicadores de la calidad microbiológica aislados a partir de muestras de: (i) agua de 16 canales de riego en la provincia de Tungurahua-Ecuador y (ii) los utensilios e insumos de una cocina en una cafeterí­a en Quito-Ecuador. Para esto se confrontó 4 ppm, 10 pmm y 15 ppm de agua de plata coloidal durante 15, 30 y 60 minutos con los crecimientos equivalentes a A625 de 0,08-0,1 de las cepas de: Escherichia coli ATCC 11229, Salmonella Choleraesuis ATCC 10708, Staphylococcus aureus ATCC 6538 y Pseudomonas aeruginosa ATCC 15442. Una vez estandarizada la técnica se comparó la densidad poblacional de: mesófilos aerobios, E. coli -coliformes totales y enterobacterias de 16 muestras de agua de canales de riego y de 11 superficies de preparación de alimentos antes y después de la exposición a 4 ppm, 10 ppm y 15 ppm de la suspensión de plata coloidal. La plata inhibió el crecimiento de todos los grupos microbianos evaluados en los canales de riego excepto en el 31,3% (n=5) de los casos en los que a 4 ppm de plata los recuentos de las bacterias mesófilas aerobias estuvieron en un rango de entre 10x102 UFC/mL y 40x102 UFC/mL. Para el caso de las muestras recolectadas en la cafeterí­a, los recuentos de mesófilos aerobios disminuyeron en el 64,3% (n=9) del total de superficies muestreadas mientras que el número de E. coli - coliformes totales disminuyó en el 93% (n=13) de las áreas muestreadas a partir los 5 minutos de exposición con de 4 ppm de agua de plata coloidal.

Tandem Autologous-Autologous Vs. Autologous-Allogeneic Transplantation for Newly Diagnosed Multiple Myeloma: Pooled Analysis of 1,338 Patients from Four Trials with Long-Term Follow up

Introduction: Tandem autologous transplant (auto- auto) has been studied as a method to increase remission rates and reduce relapse in the upfront therapy of MM. The use of autologous followed by reduced intensity conditioning allogenic transplantation (auto-allo) offers the potential of long-term graft-versus-myeloma (GVM) effect, but with the risk of graft versus host disease and potentially higher non-relapse mortality (NRM). Trials comparing these two strategies relied on availability of HLA-matched sibling donors for arm allocation (biological "randomization") and have yielded conflicting results, in part due to trial size or limited follow up. A pooled analysis of multiple trials with extended follow up provides the best opportunity to compare these two transplant strategies. Methods: We obtained individual patient data from participants of 4 trials comparing auto-auto vs. auto-allo after brief induction therapy, namely BMT CTN 0102 (N=709), NMAM2000 (N=357), PETHEMA/GEM2000 (N=110), and the Torino consortium trial (N=162). In all 4 trials arm allocation was by biological "randomization". Patients were designated high risk if beta-2 microglobulin ≥ 4.0 mg/L at diagnosis or presence of deletion of chromosome 13 by metaphase karyotyping. Time to event outcomes were analyzed by intention to treat, from the time of first autologous transplant. Main outcomes analyzed were overall survival (OS) and progression free survival (PFS). Secondary outcomes analyzed were NRM and risk of relapse, treated as competing risks, and post relapse survival. Results: There were 1,338 patients included in the analysis, 899 in auto-auto and 439 in auto-allo. Median follow up of survivors is 118.5 months. Characteristics of the two arms are displayed in Table 1. Median OS was 78.0 months in auto-auto and 98.3 months in auto-allo (HR= 0.85, P=0.003, Figure 1). OS was 59.8 % vs. 62.3% at 5-years (P=0.37) and 36.4% vs. 44.1% at 10 years (P=0.01) for auto-auto and auto-allo respectively. PFS was also improved in auto-allo (HR= 0.84, P=0.004) with 5-year PFS of 23.4 vs. 30.1% (P=0.01) and 10-year PFS of 14.4% vs. 18.7% (P=0.06). For the 214 high risk patients (125 auto-auto, 89 auto-allo) there was superior 5-year and 10-year PFS with auto-allo, but no difference in OS. Risk of NRM was higher in auto-allo (10 year 8.3% vs. 19.7%, P&amp;lt;0.001), while risk of disease progression was higher in auto-auto (10 year 77.2% vs. 61.6%, P&amp;lt;0.001). There were 685 progressions in auto-auto and 266 in auto-allo. Median post relapse overall survival was 41.5 months in auto-auto and 62.3 months in auto-allo (HR= 0.71, P&amp;lt;0.001, Figure 2). Five years post relapse, 37.0% of patients were alive in auto-auto vs. 51.1% in auto-allo (P&amp;lt;0.001). Conclusion: Long-term follow up using a large pooled dataset of 4 trials indicates durable, long-term disease control with an auto-allo strategy. Despite higher NRM, there was a reduction in the risk of relapse and superior post relapse survival in auto-allo. This supports the hypothesis of a durable GVM effect enhancing myeloma control with subsequent therapies. Disclosures Costa: Janssen: Research Funding, Speakers Bureau; Amgen: Consultancy, Honoraria, Research Funding, Speakers Bureau; Celgene: Consultancy, Honoraria, Research Funding; Abbvie: Consultancy; Karyopharm: Consultancy; Fujimoto Pharmaceutical Corporation Japan: Other: Advisor; Sanofi: Consultancy, Honoraria, Speakers Bureau; GSK: Consultancy, Honoraria, Research Funding. Pasquini:Pfizer: Consultancy; Medigene: Consultancy; Amgen: Consultancy; Novartis: Research Funding; Kite Pharmaceuticals: Research Funding; BMS: Research Funding. Bladé:Janssen, Celgene, Amgen, Takeda: Membership on an entity's Board of Directors or advisory committees; Irctures: Honoraria. Schonland:Sanofi: Research Funding; Takeda: Honoraria, Research Funding; Prothena: Honoraria; Medac: Other: Travel grant; Janssen: Honoraria, Membership on an entity's Board of Directors or advisory committees, Research Funding. Hari:Celgene: Consultancy, Honoraria, Research Funding; Takeda: Consultancy, Honoraria, Research Funding; BMS: Consultancy, Research Funding; Janssen: Consultancy, Honoraria; Kite: Consultancy, Honoraria; Amgen: Research Funding; Spectrum: Consultancy, Research Funding; Sanofi: Honoraria, Research Funding; Cell Vault: Equity Ownership; AbbVie: Consultancy, Honoraria. Giralt:Jazz Pharmaceuticals: Consultancy; Novartis: Consultancy; Takeda: Consultancy, Research Funding; Celgene: Consultancy, Research Funding; Kite: Consultancy; Johnson &amp; Johnson: Consultancy, Research Funding; Actinium: Consultancy, Research Funding; Miltenyi: Research Funding; Spectrum Pharmaceuticals: Consultancy; Amgen: Consultancy, Research Funding. Patriarca:Takeda: Membership on an entity's Board of Directors or advisory committees; Celgene: Membership on an entity's Board of Directors or advisory committees; Janssen: Membership on an entity's Board of Directors or advisory committees. Stadtmauer:Abbvie: Research Funding; Janssen: Consultancy; Amgen: Consultancy; Novartis: Consultancy, Research Funding; Tmunity: Research Funding; Celgene: Consultancy; Takeda: Consultancy. Krishnan:Celgene, Janssen, Sanofi, BMS: Consultancy; Sutro BioPharma, zPredicta: Consultancy; Amgen, Takeda: Speakers Bureau; Celgene, Z Predicta: Other: Stock Ownership; Takeda: Research Funding.

A Prospective Analysis of the Genomic Landscape of Patients with Acute Myeloid Leukemia and Its Impact on Clinical Outcomes - Data from a Tertiary Care Center in India

BACKGROUND : Clinical Phenotype and outcomes of patients with Acute myeloid leukemia (AML) in the Indian subcontinent differs from published literature. A younger age at diagnosis and higher induction mortality complicate AML management in India(1). Metaphase Karyotyping represents the backbone of prognostication and risk stratification in AML. Optimal treatment strategies for the cohort of Cytogenetically normal AML are still under evaluation. Applications of Next generation Sequencing (NGS) techniques in AML have unravelled the genetic heterogeneity of this disease. Whole genome sequencing has identified many novel mutations leading to tremendous improvements in diagnosis and risk stratification. Development of therapies targeting these genetic alterations is enabling a gradual shift from non-specific approaches to personalised therapy tailored to an individual patient's genome. This will undoubtedly translate to better clinical outcomes for this disease, with otherwise poor prognosis. Whole genome sequencing is still in a nascent stage in Indian settings with no published literature on genomics in AML till date. We aimed to study the genomic landscape of AML in the Indian population and to co-relate this with clinical outcomes over the course of 1 year. METHODS: We recruited 34 newly diagnosed patients with AML who presented to our Centre (Mazumdar Shaw Medical Centre, Narayana Health City, Bangalore, India) between November 2017 and May 2018. Clinical and laboratory details of all patients were recorded. Bone marrow and paired peripheral blood samples were drawn before initiating therapy. Whole genome sequencing and Exome capture was done for each sample using Ilumina HiSeq platform. Patients were risk stratified as per ELN 2017 and treated as per NCCN guidelines. Patients were followed up prospectively for one year from initial diagnosis. Genetic results were stratified according to gene function and analysed with respect to predefined clinical outcomes (remission status post induction, relapse rates, progression free and overall survival). RESULTS: Amongst the 34 study participants, 5 patients failed QC during sequencing and were de-recruited. Hence 29 patients were available for final analysis. Median age of patients was 42 years with 13 patients (44.8%) less than 40 years of age.18 patients (60%) had normal cytogenetics at baseline.17 patients (58%) were classified as intermediate risk and 6 patients each as Standard and high risk, as per ELN 2017. 22 patients (79.3%) patients received standard Induction chemotherapy (3+7 regimen) while 6 patients received hypomethylating agents. Overall CR rate following induction at Day 28 was 50% and Induction mortality was 21.42%. 6 patients underwent an Allogenic Stem cell transplant. A total of 96 mutations (47 driver and 49 VUS mutations) in 123 genes were identified. The average number of Driver mutations was 1.48 per patient. IDH genes were the most frequently mutated Driver genes followed by FLT3 mutations. Frequency of NPM1 mutations was significantly low (17.25%). Highest frequency of VUS mutations was seen in the ETV6, ATM and CBLC genes. Highest frequency of somatic mutations were identified in the genes encoding for myeloid transcription factors and DNA methylation. Average driver mutations showed significant co-relation to Age (&gt; 60 years) and high burden of Bone marrow blasts (&gt;30%). An updated risk stratification incorporating mutation analysis findings resulted in re-stratification of 8 intermediate risk patients into high risk. 2 patients with detectable FLT3 ITD mutation by NGS were negative by PCR. Choice of consolidation therapy and Driver mutation status were found to show statistically significant association with both Event free survival and Overall survival at 1 year. Increased driver mutation burden was associated with increased refractoriness to chemotherapy and poor EFS and OS. Mutations in Tumour suppressor genes, were associated with suboptimal treatment outcomes and poor survival. CONCLUSIONS Genomic landscape of AML in Indian patients shows significant differences from published literature. This may hold clues to the differing biological characteristics of AML seen in this population. Genome based risk stratification and tailored therapy needs to be adapted into the management of AML. This data provides valuable insights into developing therapeutic strategies for Indian patients. Disclosures No relevant conflicts of interest to declare.

Metaphase Cytogenetics for Risk Stratification in Newly Diagnosed Multiple Myeloma

Introduction: Abnormal metaphase cytogenetics are associated with inferior survival in newly diagnosed multiple myeloma (MM). These abnormalities are only detected in one third of cases due to the low proliferative rate of plasma cells. It is unknown if metaphase cytogenetics improve risk stratification when using contemporary prognostic models such as the revised international staging system (R-ISS), which incorporates interphase fluorescence in situ hybridization (FISH). Aims: The aims of this study were to 1) characterize the association between abnormalities on metaphase cytogenetics and overall survival (OS) in newly diagnosed MM treated with novel agents and 2) evaluate whether the addition of metaphase cytogenetics to R-ISS, age, and plasma cell labeling index (PCLI) improves model discrimination with respect to OS. Methods: We analyzed a retrospective cohort of 483 newly diagnosed MM patients treated with proteasome inhibitors (PI) and/or immunomodulators (IMID) who had metaphase cytogenetics performed prior to initiation of therapy. Abnormal metaphase cytogenetics were defined as MM specific abnormalities, while normal metaphase cytogenetics included constitutional cytogenetic variants, age-related Y chromosome loss, and normal metaphase karyotypes. Multivariable adjusted proportional hazards regression models were fit for the association between known prognostic factors and OS. Covariates associated with inferior OS on multivariable analysis included R-ISS stage, age ≥ 70, PCLI ≥ 2, and abnormal metaphase cytogenetics. We devised a risk scoring system weighted by their respective hazard ratios (R-ISS II +1, R-ISS III + 2, age ≥ 70 +2, PCLI ≥ 2 +1, metaphase cytogenetic abnormalities + 1). Low (LR), intermediate (IR), and high risk (HR) groups were established based on risk scores of 0-1, 2-3, and 4-5 in modeling without metaphase cytogenetics, and scores of 0-1, 2-3, and 4-6 in modeling incorporating metaphase cytogenetics, respectively. Survival estimates were calculated using the Kaplan-Meier method. Survival analysis was stratified by LR, IR, and HR groups in models 1) excluding metaphase cytogenetics 2) including metaphase cytogenetics and 3) including metaphase cytogenetics, with IR stratified by presence and absence of metaphase cytogenetic abnormalities. Survival estimates were compared between groups using the log-rank test. Harrell's C was used to compare the predictive power of risk modeling with and without metaphase cytogenetics. Results: Median age at diagnosis was 66 (31-95), 281 patients (58%) were men, median follow up was 5.5 years (0.04-14.4), and median OS was 6.4 years (95% CI 5.7-6.8). Ninety-seven patients (20%) were R-ISS stage I, 318 (66%) stage II, and 68 (14%) stage III. One-hundred and fourteen patients (24%) had high-risk abnormalities by FISH, and 115 (24%) had abnormal metaphase cytogenetics. Three-hundred and thirteen patients (65%) received an IMID, 119 (25%) a PI, 51 (10%) received IMID and PI, and 137 (28%) underwent upfront autologous hematopoietic stem cell transplantation (ASCT). On multivariable analysis, R-ISS (HR 1.59, 95% CI 1.29-1.97, p &lt; 0.001), age ≥ 70 (HR 2.32, 95% CI 1.83-2.93, p &lt; 0.001), PCLI ≥ 2, (HR 1.52, 95% CI 1.16-2.00, p=0.002) and abnormalities on metaphase cytogenetics (HR 1.35, 95% CI 1.05-1.75, p=0.019) were associated with inferior OS. IR and HR groups experienced significantly worse survival compared to LR groups in models excluding (Figure 1A) and including (Figure 1B) the effect of metaphase cytogenetics (p &lt; 0.001 for all comparisons). However, the inclusion of metaphase cytogenetics did not improve discrimination. Likewise, subgroup analysis of IR patients by the presence or absence of metaphase cytogenetic abnormalities did not improve risk stratification (Figure 1C) (p &lt; 0.001). The addition of metaphase cytogenetics to risk modeling with R-ISS stage, age ≥ 70, and PCLI ≥ 2 did not improve prognostic performance when evaluated by Harrell's C (c=0.636 without cytogenetics, c=0.642 with cytogenetics, absolute difference 0.005, 95% CI 0.002-0.012, p=0.142). Conclusions: Abnormalities on metaphase cytogenetics at diagnosis are associated with inferior OS in MM when accounting for the effects of R-ISS, age, and PCLI. However, the addition of metaphase cytogenetics to prognostic modeling incorporating these covariates did not significantly improve risk stratification. Disclosures Lacy: Celgene: Research Funding. Dispenzieri:Akcea: Consultancy; Intellia: Consultancy; Alnylam: Research Funding; Celgene: Research Funding; Janssen: Consultancy; Pfizer: Research Funding; Takeda: Research Funding. Kapoor:Celgene: Honoraria; Sanofi: Consultancy, Research Funding; Janssen: Research Funding; Cellectar: Consultancy; Takeda: Honoraria, Research Funding; Amgen: Research Funding; Glaxo Smith Kline: Research Funding. Leung:Prothena: Membership on an entity's Board of Directors or advisory committees; Takeda: Research Funding; Omeros: Research Funding; Aduro: Membership on an entity's Board of Directors or advisory committees. Kumar:Celgene: Consultancy, Research Funding; Janssen: Consultancy, Research Funding; Takeda: Research Funding.