Cytogenetics Laboratory Research Articles

Development of a Chromosomal Microarray Test for the Detection of Abnormalities in Formalin-Fixed, Paraffin-Embedded Products of Conception Specimens

Testing the products of conception (POCs) provides information about the cause of fetal loss and helps determine the recurrence risk of future losses and chromosome abnormalities in subsequent pregnancies. Historically, the Mayo Clinic Cytogenetics Laboratory performed targeted fluorescent in situ hybridization (FISH) testing to identify aneuploidy of only certain chromosomes in formalin-fixed, paraffin-embedded (FFPE) POC samples. Chromosomal microarray studies using the Affymetrix OncoScan FFPE Assay can detect copy number changes across the genome. We validated the utility of the OncoScan assay using 25 archival FFPE POC specimens with previous FISH results (five normal, 12 trisomy, six triploidy, two monosomy). Of the five normal samples, four had no clinically relevant findings, and one sample was found to have trisomy 9, which is not detectable by the FISH test. For the 20 samples with abnormal FISH results, the OncoScan assay identified all reported abnormalities along with additional findings. A sample with trisomy 22 was also found to have trisomy 7. Another sample reported as triploidy was found to have four copies of chromosome 16. In conclusion, we verified the performance characteristics of OncoScan on FFPE POC specimens and found it acceptable for clinical use. Additional information was identified in 3 of 25 cases (12%) that would explain the pregnancy loss or provide recurrence risk for the family.

The incidence of acute myeloid leukemia in Calgary, Alberta, Canada: a retrospective cohort study

BackgroundThe incidence rate of acute myeloid leukemia (AML) was determined in the Calgary Metropolitan Area, a major Canadian city.MethodsData from all patients diagnosed with AML between January 1, 2011 and December 31, 2015 were retrieved from a single, centralized cancer cytogenetics laboratory for bone marrow samples, the sole diagnostic facility of its kind in Southern Alberta.ResultsThe calculated incidence rate was 2.79 cases per 100,000 person-years with a median age of 60, slightly lower than previously published data. The age-standardized incidence rate for Canada was 3.46 cases per 100,000 person-years. The higher value is reflective of Calgary’s younger population compared to the rest of Canada. Higher male incidence and greatest incidence occurring at approximately the age of 85 is similar to data from other developed countries. The lower incidence rates and median age of diagnosis, in comparison with that of other high-income nations, may be due to differences in the proportion of aging citizens in the population.ConclusionThis is the first published incidence rate of acute myeloid leukemia (AML) in Canada across all age groups.

Diagnostic cytogenetic testing following positive noninvasive prenatal screening results: a clinical laboratory practice resource of the American College of Medical Genetics and Genomics (ACMG).

Disclaimer: ACMG Clinical Laboratory Practice Resources are developed primarily as an educational tool for clinical laboratory geneticists to help them provide quality clinical laboratory genetic services. Adherence to these practice resources is voluntary and does not necessarily assure a successful medical outcome. This Clinical Laboratory Practice Resource should not be considered inclusive of all proper procedures and tests or exclusive of other procedures and tests that are reasonably directed to obtaining the same results. In determining the propriety of any specific procedure or test, the clinical laboratory geneticist should apply his or her own professional judgment to the specific circumstances presented by the individual patient or specimen. Clinical laboratory geneticists are encouraged to document in the patient's record the rationale for the use of a particular procedure or test, whether or not it is in conformance with this Clinical Laboratory Practice Resource. They also are advised to take notice of the date any particular guideline was adopted, and to consider other relevant medical and scientific information that becomes available after that date. It also would be prudent to consider whether intellectual property interests may restrict the performance of certain tests and other procedures.Noninvasive prenatal screening (NIPS) using cell-free DNA has been rapidly adopted into prenatal care. Since NIPS is a screening test, diagnostic testing is recommended to confirm all cases of screen-positive NIPS results. For cytogenetics laboratories performing confirmatory testing on prenatal diagnostic samples, a standardized testing algorithm is needed to ensure that the appropriate testing takes place. This algorithm includes diagnostic testing by either chorionic villi sampling or amniocentesis samples and encompasses chromosome analysis, fluorescence in situ hybridization, and chromosomal microarray.

A novel deletion of SNURF/SNRPN exon 1 in a patient with Prader-Willi-like phenotype

Here we report the smallest deletion involving SNURF/SNRPN that causes major symptoms of Prader-Willi syndrome (PWS), including hypotonia, dysmorphic features, intellectual disability, and obesity. A female patient with the aforementioned and additional features was referred to the Mayo Clinic Cytogenetics laboratory for genetic testing. Chromosomal microarray analysis and subsequent Sanger sequencing identified a de novo 6.4 kb deletion at 15q11.2, containing exon 1 of the SNURF gene and exon 1 of the shortest isoform of the SNRPN gene. SNURF/SNRPN exon 1, which is methylated on the silent maternal allele, is associated with acetylated histones on the expressed paternal allele. This region also overlaps with the PWS-imprinting center (IC). Subsequent molecular methylation analysis was performed using methylation-specific MLPA (MS-MLPA), which characterized that the deletion of SNURF/SNRPN exon 1 was paternal in origin, consistent with the PWS-like phenotype. Since SNURF/SNRPN gene and the PWS-IC are known to regulate snoRNAs, it is likely that the PWS-like phenotype observed in patients with paternal SNURF/SNRPN deletion is due to the disrupted expression of SNORD116 snoRNAs.

PRENATAL DIAGNOSIS WITH GENETIC COUNSELING IN UHC SPLIT, CROATIA.

27Mar 2017 PRENATAL DIAGNOSIS WITH GENETIC COUNSELING IN UHC SPLIT, CROATIA. Robert Vulic , Vida Culic , Silvana Miskovic , Ivana Gunjaca , Damir Roje , Ruzica Lasan and Jasminka Pavelic. Gynecology and Obstetrics Outpetient Clinic, Split, Croatia. Department for Medical Genetics with Laboratory for Human Genetics and Genetic Counseling Unit, Paediatrics Clinic, University Hospital Centre Split, Croatia. School of Medicine, University of Split, Split, Croatia. Gynecology and Obstetrics Clinics, University Hospital Centre Split, University of Split, School of Medicine, Split, Croatia. Cytogenetic Laboratory, Paediatrics Clinic, Clinical Hospital Centre Zagreb, Croatia. Institute Ru?er Boškovi?, Division of Molecular Medicine, Zagreb, Croatia.

Estimates of live birth prevalence of children with Down syndrome in the period 1991-2015 in the Netherlands.

In Western countries, increasing maternal age has led to more pregnancies with a child with Down syndrome (DS). However, prenatal screening programs, diagnostic testing and termination of pregnancy influence the actual DS live birth (LB) prevalence as well. The aim of this study is to examine these factors in the Netherlands for the period 1991-2015. In our study, we establish a baseline for DS LB prevalence before non-invasive prenatal testing will be made available to all pregnant women in the Netherlands in 2017. Full nationwide data from the Dutch cytogenetic laboratories were used to evaluate the actual DS LB prevalence. In addition, nonselective DS prevalence, which is the DS LB prevalence that would be expected in absence of termination of pregnancies, was estimated on the basis of maternal age distribution in the general population. Because of an increase in maternal age, nonselective DS prevalence increased from around 15.6 [95% confidence interval (CI) 13.9-17.4] per 10000 LBs in 1991 (311 children in total) to around 22.6 (95% CI 20.3-24.9) per 10000 in 2015 (385), the increase levelling off in recent years. Actual LB prevalence rose from around 11.6 (95% CI 10.9-12.2) per 10000 in 1991 (230 children) to an estimated peak of 15.9 (95% CI 15.6-16.2) per 10000 in 2002 (322), gradually decreasing since to 11.1 (95% CI 10.8-11.5) per 10000 in 2015 (190). Reduction of DS LBs resulting from elective terminations had been fairly constant between 1995 and 2002 at around 28% and rose afterwards from 35% in 2003 to around 50% in 2015. In spite of expansion of antenatal screening in the Netherlands in the 1990s and early 2000s, actual DS LB prevalence increased during this period. However, after 2002, this trend reversed, probably because of informing all pregnant women about prenatal testing since 2004 and the implementation of a national screening program in 2007.

Analyse du parcours de dépistage prénatal des femmes ayant donné naissance en France à un enfant atteint de trisomie 21

ObjectiveThe main objective of this study was to screen the prenatal follow-up of women with live birth trisomy 21 child in order to evaluate the proportion of prenatal screening failure versus cases where the women refused either the screening or the prenatal diagnosis of Down syndrome. This study covers the period of time from 2009 to 2012 when the national prenatal screening policy changed from second to first trimester and allows for a comparative assessment of the nationwide efficiency of the various maternal serum marker based strategies. MethodAll authorized cytogenetic laboratories sent required data for all cases of trisomy 21 diagnosed in FRANCE in new-borns (less than 1-year-old) from January 2010 to July 2013. ResultsA total of 1253 cases of trisomy 21 were diagnosed before 1 year of age whose mother did not had prenatal diagnosis. For 861 of them, information on the prenatal follow-up was available, with 72% of cases where a prenatal screening was organized either by maternal serum marker or by ultrasound. Results of the screening strategy was positive with maternal serum marker in 28% of cases (calculated risk≥1/250), positive because of abnormal ultrasound in 5% and negative with maternal marker screening (whatever the strategy used) in 67% of cases. Detection rate over the period of the study was 82%, with similar efficiency of first and second trimester strategies (83%) but significantly lower with sequential association of first trimester Nuchal translucency measurement and second trimester serum screening (70%). ConclusionSwitching from second trimester to first trimester screening strategy, with as many trisomy 21 foetuses diagnosed with half invasive procedures fulfilled national health policy objectives. Analysis of these data gives useful insights to elaborate a future screening policy involving cell-free foetal DNA sequencing.

Pre- and Postnatal Genetic Evaluation reduce the Reproductive Risk of nonhomologous Robertsonian Translocation Carrier Couple

ABSTRACTPurposeSpontaneous abortion has been reported in 15 to 20% of all diagnosed pregnancies. The most common cause of spontaneous abortion is chromosomal abnormalities of the embryo. Robertsonian translocation (RT) is one of the major chromosomal rearrangements with a prevalence rate of 0.1% of the general population and 1% of the infertile population. Robertsonian translocation carriers, especially 21-14, are the most common balanced rearrangement among the carrier couples with a history of spontaneous abortion.Materials and methodsCytogenetic analysis was carried out based on phytohemagglutinin-stimulated peripheral blood lymphocyte cultures and without phytohemagglutinin-stimulated amniocyte culture. Lymphocyte and amniocyte culturing and GTG banding were performed following standard protocols as described by the Association of genetic technologists (AGT) Cytogenetics Laboratory Manual.ResultsCytogenetic evaluation of both the partners and the child revealed that the child had translocated Down's syndrome and the mother was a carrier of balanced RT of 14q;21q. Amniocentesis of the next pregnancy and detection of chromosomal abnormality in the fetus was done by fluorescencein situhybridization (FISH) analysis of the amniotic cells with 13,18,21,X,Y probe mix found normal chromosomal constituent in the fetus.ConclusionThe present study shows that genetic counseling, cytogenetic evaluation, prenatal diagnosis by amniocentesis, and FISH together help couples with nonhomologous RT and history with syndromic child and repeated abortions to get normal offspring.How to cite this articleDe P, Chakravarty S, Chakravarty A. Pre- and Postnatal Genetic Evaluation reduce the Reproductive Risk of nonhomologous Robertsonian Translocation Carrier Couple. Int J Infertil Fetal Med 2017;8(1):36-40.

The laboratories of anatomy and the standard practices therein

A laboratory is a facility that is equipped for carrying out scientific experiments or procedures for the purposes of research, teaching, or analysis for service provision under controlled and optimal working conditions. Anatomy is the basic essential science of medicine, laboratories of anatomy are those facilities equipped for carrying out such scientific experiments to understand the structures and functions of human and animal bodies. The laboratories of anatomy, as generally required, for any conventional laboratory, do serve three known basic functions. Each could be a teaching, research, or service laboratory or perform two or three of the functions. Common laboratory units in the department of anatomy include anatomy museum and workshop, animal cell and tissue culture laboratory, biological anthropology and forensic anatomy laboratory, clinical embryology and andrology laboratory, cytogenetic laboratory, histology and histochemistry laboratory, mortuary and gross anatomy laboratory, and radiological anatomy laboratory.

Cutaneous Manifestations of Fanconi's Anemia in Two Siblings.

Sir, Fanconi's anaemia is a rare, autosomal recessive disorder characterized by diverse congenital malformations, developmental defects, progressive pancytopenia, predisposition to both hematologic malignancies and solid tumors, and widespread mottled skin pigmentation. Two brothers (aged 7 and 13 years) born of nonconsanguinous marriage and studying in second and sixth standard presently, presented to our pediatrics department 2 years back with the complaints of fever, not gaining weight and height, paleness of body, as well as an additional complaint of incomplete voiding of urine in the younger brother. On anthropometric examination, the weight of both the brothers was less than the 3rd centile. The height of the elder brother was at the 3rd centile and younger brother was less than the 3rd centile. The head, chest, and mid arm circumference was 48 cm, 49 cm, and 14.5 cm, respectively, for the elder brother and 47.5 cm, 46 cm, and 12cm, respectively, for the younger brother. Microcephaly and micrognathia was present in the younger brother. On hematological evaluations, there was progressive pancytopenia. Peripheral blood picture showed moderate anisopoikilocytosis with microcytic hypochromic anemia in both the brothers. Bone marrow aspirate examination revealed hypoplastic anemia. Liver function, renal function tests, and X-rays of the hands were normal. On ultrasonography of the abdomen, there was hepatomegaly, and cystitis with a large echogenic calculus of size 1.2 cm in the dependent part of urinary bladder in the elder and younger brother, respectively. Karyotyping done from a cytogenetics lab, Department of Zoology was normal. The patients were referred to the dermatology outpatient department for the cutaneous manifestations. On general examination, there was pallor, generalized olive brown pigmentation of the whole body, with discrete, asymptomatic, hypopigmented macules distributed over the trunk [Figures 1 and 2], neck, bilateral upper limbs, lower limbs, face, palms [Figure 3], and soles in both the brothers. No cafe-au-lait macules, palmoplantar keratoderma, and mucosal pigmentation was present. Fingers, thumb and nails, and testes were normal. No hearing loss, structural abnormalities of external ears, and squint were found. Fundoscopy, superficial, and deep reflexes were normal. Histopathological examination of the skin revealed sparse superficial perivascular lymphocytic infiltrate with numerous melanophages within papillary dermis. The papillary dermis was slightly thickened and showed delicate fibroplasia and mucin with flattened epidermis at places [Figure 4].Figure 1: Mottled skin pigmentation on the trunkFigure 2: Siblings having “raindrop pigmentation” on the trunkFigure 3: Mottled pigmentation on the palmsFigure 4: Histopathology from the trunk showing flattened epidermis, slightly thickened papillary dermis, with numerous melanophages on hematoxylin and eosin stain (×10)Fanconi anemia is a genetically and phenotypically heterogeneous recessive disorder.[1] There is an intrafamilial phenotypic variation in the specific types of congenital malformations among affected siblings.[2] Other findings include skin pigmentation abnormalities, hypoplastic thenar eminence, microcephaly, and/or microphthalmia.[1] Renal aplasia, horseshoe kidney, and syndactyly have also been reported.[3] However, in our case, only cystitis and large echogenic calculus of size 1.2 cm was present in the dependent part of the urinary bladder, in the younger brother. In our patients, mottled pigmentation all over the body, short stature with height and weight below the 3rd centile, microcephaly in the younger brother, progressive pancytopenia with hypoplastic anaemia on bone marrow aspiration, led to the diagnosis of Fanconi's anemia. The patients were given blood transfusions and anabolic steroids (stanozolol 2 mg/kg/day initially which is still continuing). They are on continuous follow up. There is weight gain, increase in height, and improvement in blood counts. Declaration of patient consent The authors certify that they have obtained all appropriate patient consent forms. In the form the patient(s) has/have given his/her/their consent for his/her/their images and other clinical information to be reported in the journal. The patients understand that their names and initials will not be published and due efforts will be made to conceal their identity, but anonymity cannot be guaranteed. Financial support and sponsorship Nil. Conflicts of interest There are no conflicts of interest. Acknowledgement We acknowledge the assistance of Dr. Raman Lab, Department of Zoology, Banaras Hindu University for karyotyping.

Cytogenetics, Molecular Genetics and Epigenetics and Their Impact on The Management of Acute Lymphoblastic Leukemia

The recent improvements in the outcomes of patients with acute lymphoblastic leukemia reflect the progress in the field of diagnostics and the achievements in therapeutic interventions. In particular, the availability of more advanced technology in the cytogenetic and molecular laboratories has resulted in the stratification of patients into specific risk groups and thus several novel agents as well as targeted therapies have been introduced. Additionally, precision medicine will be applicable in the near future and thus the recent developments will facilitate the provision of safer modalities of therapy that may ultimately yield not only higher responses but also improved survival rates. This review represents a recent update on the role of various cytogenetic assays and molecular techniques in patients with acute lymphoblastic leukemia. It is composed of a number of sections including: history of cytogenetics, disease etiology and pathophysiology, utilization of various cytogenetic assays diagnostically, disease-specific cytogenetic and molecular abnormalities, common disease genetic subtypes, prognosis and risk stratification, refractory and relapsed disease, novel therapies, precision medicine and drug repurposing.

Four Novel Balanced Reciprocal Translocations and a Novel Inversion Associated with Recurrent Spontaneous Abortions- A case report.

Summary Aim/background The main objective of this study was to evaluate the cause of reproductive failures in couples by clinical and cytogenetic analysis. Couples with history of recurrent pregnancy losses (RPL) were referred to our cytogenetic laboratory for chromosomal evaluation. Methods Chromosomal analysis from lymphocyte culture of the phenotypically normal couples were done to evaluate the role of chromosomal abnormalities in Recurrent spontaneous abortion (RSA) and to offer appropriate genetic counseling to them. Results Clinical and hormonal profile of the couples revealed normal phenotypes. Chromosomal analysis of the couples showed karyotypes like 45,XX,der(13;14)(q10;q10), 46,XX,t(1;17)(p36.2;p13.1), 46,XX,t(9;16)(p24.2;q12.2), 46,XX,t(1;18)(q42.1;q21.2), 46,XY,inv(9)(p11.2q13.3), 46,XY,inv(Y)(p11.2q11.223), 46,XX,inv(14)(q23.2q32.1), 46,XY,(16qh+), 46,XY,t(4;18)(q13.1;q23.0), 46,XX,t(12;14)(q24.1;q32.2), inv(11)(q23.2q24.3). Conclusion Among the cases mentioned in this study four translocation cases and inversion and translocation within single individual were not previously reported in any other literature. Novel balanced reciprocal translocations (BRT) were reported as original investigation in four individuals and single case of translocation and inversion within same individual in new break-points. The study showed chromosomal anomalies like balanced translocation, inversion and duplication causing recurrent pregnancy losses from different unrelated families and there was no history of consanguineous marriage in any of them.

CHROMOSOMAL MUTAGENESIS IN HUMAN SOMATIC CELLS: 30-YEAR CYTOGENETIC MONITORING AFTER CHORNOBYL ACCIDENT

In the lecture we have generalized and analyzed the data of cytogenetic laboratory of National Research Center for Radiation Medicine of the National Academy of Medical Sciences of Ukraine on 30-year selective cytogenetic monitoring among the priority contingents of different ages exposed to radiation after Chornobyl accident in Ukraine. It is highlighted that not only targeted but also untargeted radiation-induced cytogenetic effects should be explored, especially in delayed terms following radiation exposure. The new methodical approaches for studying "bystander effect", individual radiosensitivity, and various forms of radiation-induced chromosomal instability (delayed, hidden, transmissible) have been proposed. These approaches proved to be advantageous for analyzing cytogenetic patterns of induction and persistence of chromosomal instability in human somatic cells because of "bystander effect" and "bystander type effect". The phenomenon of positive "reverse" bystander effect has been found. The possibility of modifying the inherited individual human susceptibility to mutagenic exposure by ionizing radiation has been estimated. Finally, the association between hypersensitivity to radiation exposure and realization of oncopathology in exposed individuals has been revealed. The increased intensity of human somatic chromosomal mutagenesis was confirmed not only in the nearest but in the delayed terms following Chornobyl accident as a result of radiation-induced both targeted and untargeted cytogenetic effects. Such effects can be considered as risk factors for malignant transformation of cells, hereditary diseases, birth defects, and multifactorial somatic pathology. This article is a part of a Special Issue entitled "The Chornobyl Nuclear Accident: Thirty Years After".

Cytogenetic Nomenclature and Reporting.

A standardized nomenclature is critical for the accurate and consistent description of genomic changes as identified by karyotyping, fluorescence in situ hybridization and microarray. The International System for Human Cytogenomic Nomenclature (ISCN) is the central reference for the description of karyotyping, FISH, and microarray results, and provides rules for describing cytogenetic and molecular cytogenetic findings in laboratory reports. These laboratory reports are documents to the referring clinician, and should be clear, accurate and contain all information relevant for good interpretation of the cytogenetic findings. Here, we describe guidelines for cytogenetic nomenclature and laboratory reports for cytogenetic testing applied to tumor samples.

Recurrent Cytogenetic Abnormalities in Acute Myeloid Leukemia.

The spectrum of chromosomal abnormality associated with leukemogenesis of acute myeloid leukemia (AML) is broad and heterogeneous when compared to chronic myeloid leukemia and other myeloid neoplasms. Recurrent chromosomal translocations such as t(8;21), t(15;17), and inv(16) are frequently detected, but hundreds of other uncommon chromosomal aberrations from AML also exist. This chapter discusses 22 chromosomal abnormalities that are common structural, numerical aberrations, and other important but infrequent (less than 1 %) translocations emphasized in the WHO classification. Brief morphologic, cytogenetic, and clinical characteristics are summarized, so as to provide a concise reference to cancer cytogenetic laboratories. Morphology based on FAB classification is used together with the current WHO classification due to frequent mentioning in a vast number of reference literatures. Characteristic chromosomal aberrations of other myeloid neoplasms such as myelodysplastic syndrome and myeloproliferative neoplasm will be discussed in separate chapters-except for certain abnormalities such as t(9;22) in de novo AML. Gene mutations detected in normal karyotype AML by cutting edge next generation sequencing technology are also briefly mentioned.

Chromosome Recognition.

Chromosomal analysis of human cells serves to characterize aberrations of chromosome number and structure. Individual chromosome can be identified precisely by recognition of its morphological characteristics and staining patterns according to specific landmarks, regions, and bands as described in the ideogram. Since the quality of metaphases obtained from malignant cells is generally poor for karyotyping, a practical and accurate chromosome recognition training guide is mandatory for a trainee or newly employed cytogenetic technologist in a cancer cytogenetics laboratory. The most distinguishable bands for each chromosome are described in detail in this chapter with an aim to facilitate quick and accurate karyotyping in cancers. This is an indispensable chromosome recognition guide used in a cancer cytogenetics laboratory.

Prognostic Impact of Morphology, MYC Gene Partner and BCL2/BCL6 Translocation Status in "High Grade B-Cell Lymphomas with MYC and BCL2 and/or BCL6 Rearrangements"

Introduction: "High grade B-cell lymphoma with MYC and BCL2 and/or BCL6 rearrangements" (HG DH/THL) is a new diagnosis per the revised 2016 WHO classification. It has been colloquially called "double hit/triple hit lymphoma" and is significant for its aggressive clinical course. However, under this umbrella diagnosis there are several morphologic and cytogenetic variants with either controversial or unknown clinical significance. Herein, we investigate the prognostic significance of several of these parameters, including morphology, immunoglobulin gene (IG) vs non-IG MYC translocation partners, and BCL2/BCL6 translocation status in HG DH/THL in the Mayo Clinic Rochester practice.Materials and methods: All cases, which were identified through the Mayo Clinic Lymphoma Database, University of Iowa/Mayo Clinic SPORE Molecular Epidemiology Resource (MER), and the Mayo Clinic Cytogenetics Laboratory, had diffuse large B-cell lymphoma (DLBCL) or high grade B-cell lymphoma (HGBCL) morphology, were CD20-positive, and had a MYC translocation with concurrent BCL2 and/or BCL6 translocations. Morphology was re-reviewed by four hematopathologists when possible. Cell of origin (germinal center B-cell (GCB) vs non-GCB phenotype) was determined by paraffin section immunohistochemistry using the Hans classifier. Interphase FISH was performed on either tissue paraffin sections or bone marrow aspirate smears using a breakapart probe for MYC, followed by dual fusion probes for IGH/MYC, IGL/ MYC and IGK/MYC, a BCL6 breakapart probe, and either a BCL2 breakapart probe or an IGH/BCL2 dual fusion probe. Outcome analysis was restricted to patients with de-novo HG DH/THL or HG DH/THL at transformation of a previously diagnosed low-grade lymphoma and who were treated with anthracycline-based chemotherapy. Overall survival (OS) was defined as time from DH/THL diagnosis to death from any cause.Results: There were 100 patients with a median age of 61 years (range 29-87). The consensus re-review morphologic diagnosis (n=65) was HGBCL in 39 (60%) and DLBCL in 26 (40%). The HGBCL group had inferior survival (2-year OS = 35%) compared to DLBCL (2-year OS=62%, p= 0.058). By interphase FISH, the MYC translocation partner was an IG gene (IG/MYC) in 52 cases (60%; 39 IGH, 6 IGK and 7 IGL) and a non-IG gene (non-IG/MYC) in 35 cases (40%). No difference in OS was identified between IG/MYC and non-IG/MYC cases (p=0.85, Figure 1). By FISH, 87 cases (87%) had a BCL2 translocation (59 MYC/BCL2, 20 MYC/BCL2/BCL6, and 8 MYC/BCL2/[BCL6 unknown]) and 13 (13%) lacked a BCL2 translocation (MYC/BCL6). Those with a BCL2 translocation were all of GCB phenotype (N=85), while 6 of 12 of the MYC/BCL6 cases (50%) were of non-GCB phenotype (p <0.0001). All 13 MYC/BCL6 cases were de novo diagnoses, while 38% of cases with a BCL2 translocation represented either transformation (n=22) or recurrence (n=11) events (p=0.025). The MYC/BCL6 group had better survival (2-year OS=73%) than those with BCL2 translocation (2-year OS=43%, p= 0.13).Conclusion: As HG DH/THL's frequently have morphologic characteristics of DLBCL (40% in this series), it is important to perform complete interphase FISH analysis on all cases with DLBCL or HGBCL morphology to exclude the possibility of HG DH/THL. Furthermore, within the diagnosis of HG DH/THL, there are several morphologic and cytogenetic subtypes that are biologically and clinically significant. High grade morphology (HGBCL) tends to predict for a more aggressive clinical course independent of all other parameters (p= 0.058). Unlike a recent report (Blood 2015;126:2466), our data suggest MYC translocation partner (IG versus non-IG) may have no impact on overall survival (Figure 1). HG DH/THL's with a BCL2 translocation appear restricted to GCB phenotype, frequently represent either transformation or recurrence events, and tend to have inferior OS. In contrast, those without BCL2 translocations may be of either GCB or non-GCB phenotype, typically arise de novo, and tend to have a superior OS. Careful morphologic review and complete interphase FISH studies are essential in all HG DH/THL for prognostic and potential therapeutic purposes. [Display omitted] DisclosuresNo relevant conflicts of interest to declare.

Treatment and Clinical Outcomes of High Grade B-Cell Lymphomas with MYC and BCL2 and/or BCL6 Rearrangements (Double Hit/Triple Hit Lymphomas)

Background: B-cell lymphomas with a MYC/8q24 rearrangement coupled with translocations involving BCL2 and/or BCL6, often referred to as double hit or triple hit (DH/TH) lymphomas, typically have an aggressive clinical course. A new WHO subtype, "high grade B-cell lymphoma with MYC and BCL2 and/or BCL6 rearrangements", has been established for this entity (Swerdlow et al. Blood; 127: 2375-2390, 2016). Herein we report the treatment and outcomes of DH/TH patients at our institution.Materials and Methods: Cases were identified through the Mayo Clinic Lymphoma Database, University of Iowa/Mayo Clinic SPORE Molecular Epidemiology Resource (MER), and the Mayo Clinic Cytogenetics Laboratory. Interphase FISH was performed clinically on either paraffin sections of the tissue specimens or on smears of the bone marrow aspirate specimens using break-apart probes for MYC and BCL6, dual fusion probes for IGH/MYC, IGL/MYC, and IGK/MYC, and either a BCL2 break-apart probe or an IGH/BCL2 dual fusion probe. Consensus pathology re-review was performed to define morphology as diffuse large B-cell lymphoma (DLBCL) vs. high-grade B-cell lymphoma (HGBCL). Patients with de-novo DH/TH or diagnosis of DH/TH at the time of transformation of a previously diagnosed low-grade lymphoma and who were treated with anthracycline-based chemotherapy were included in this analysis. Patients with DH/TH first identified at the time of recurrent aggressive B-cell lymphoma were excluded. Overall survival (OS) was defined as time from DH/TH diagnosis to death from any cause. EFS12 was defined as event-free status 12 months from diagnosis of DH/TH.Results: Clinical outcome analysis was performed on 71 patients (Table 1). The median age was 61 years (range 29-82). 60 patients had de-novo DH/TH and 11 patients had transformation of a previously diagnosed low-grade lymphoma. In 65 cases in which there was consensus morphology on pathology re-review, there were 39 (60%) with HGBCL morphology and 26 (40%) with DLBCL morphology. At a median follow-up of 21 months (range 1-87), the event free survival at 12 months was 43%, and the median overall survival (OS) was 22 months (95% CI: 13-NA). The OS at 5 years was 48% (95% CI: 36%-63%). Patients with DH/TH at transformation of previously diagnosed low-grade lymphoma had a very poor outcome (median OS=10.8 months, EFS12 = 10%), which was inferior to patients with de-novo DH/TH (P=0.0069). When compared to all other therapies, patients treated with R-CODOX-M/IVAC had better EFS12 (p=0.03) with a trend in improved OS (p=0.08) though this may be confounded by patient selection bias as these patients were significantly younger (p=0.0001) than patients treated with other anthracycline based regimens (table 1).Conclusion: In "high-grade B-cell lymphoma with MYC and BCL2 and/or BCL6 rearrangements", induction failure occurs early. Although transformation to DH/TH from previously diagnosed low-grade lymphoma is associated with a particularly poor outcome, the overall survival for the entire cohort at 5 years was 48%, suggesting long-term cures in a subset of patients. Treatment with R-CODOX-M/IVAC may result in superior outcomes in patients less than 60 years of age. [Display omitted] [Display omitted] DisclosuresAnsell:BMS, Seattle Genetics, Merck, Celldex and Affimed: Research Funding. Nowakowski:Celgene: Research Funding; Morphosys: Research Funding; Bayer: Consultancy, Research Funding.

Treatment Patterns and Clinical Outcomes in High-Risk Multiple Myeloma Patients Carrying the 17p Deletion: A Multi-Center Retrospective Observational Study

IntroductionImprovement in overall survival (OS) is seen primarily within standard risk Multiple Myeloma (MM), however, high risk MM OS was still around 2-3 years until recently. Del17p is a genomic imbalance which includes deletion of the TP53 locus. It occurs in ~7-10% of MM at diagnosis and is associated with extramedullary disease and is a strong poor prognostic factor. Recently approved novel agents and combinations have demonstrated improved outcomes also in patients with negative cytogenetic features; however, their long term impact remains to be seen. Access to newer agents depends on time of diagnosis as well as availability of clinical trials, access programs and insurance coverage. The goal of this study is to analyze real world data including treatment patterns and outcomes among MM patients carrying 17p deletion, in a retrospective multi-site study.MethodsAn observational, retrospective, multi-center study. Consecutive patients diagnosed with multiple myeloma in the 8 participating centers in Israel, diagnosed between 1.1.2008 - 3.1.2016 that were proven to carry 17p deletion by means of FISH studies (any % of plasma cells), were identified by searching hospital records including cytogenetic lab records. Data concerning patient demographics, disease characteristics, treatment regimens and clinical endpoints were collected.ResultsA total of 57 patients carrying 17p deletion of FISH were identified. Patient's characteristics are described in table. Notably, most patients had bone disease; extramedullary disease (EMD) rate was relatively high, as was the presence of additional high risk FISH abnormalities. Most patients received a bortezomib-based induction, over half underwent ASCT. Fifteen (26%) of the patients participated in clinical trials or access programs (or both) (table). Overall response rate (ORR) after induction was 84%, response rate declined in subsequent treatment lines (figure 1). The median follow-up was 21 (range, 4-94) months. Median overall survival (OS) was 43 months; Median Progression free survival (PFS) was 20 months (figure 2A,B). In univariate analysis, presence of extra-medullary disease at diagnosis was associated with worse PFS (7.4 vs. 21.4 months, p=0.05, figure 2C); presence of additional high risk FISH findings also trended towards shorter PFS (13.5 vs 21.4 months, p=0.0569), while age, gender, ISS, %plasma cells, % cells with del17p, time of del17p detection, M-Spike and iFLC levels were not significantly associated with PFS. Among patients with a PFS greater than 6 months, ASCT was associated with a significant improvement in PFS (25.7 vs 9.0 months, p=0.0022 log-rank test) (figure 2D), 4 patients underwent allotransplant, with a median OS of 69 months.ConclusionsOur data confirm the poor prognosis of myeloma patients with del17p, in a multi-site observational setting, and an even worse prognosis in the presence of extramedullary disease and additional high-risk FISH features. While ORR after induction is similar to that generally expected in a newly diagnosed myeloma patients, responses are shorter and thus PFS is inferior compared to recently reported upfront bortezomib-based regimens ranging from 30-40+ months. Responses decline further in subsequent lines yet remain non-negligible even in advanced therapeutic lines. Our data support the role of ASCT in these high risk patients, and the potential role of allotransplant in selected patients. High rate of participation in clinical trials and access programs throughout the course of therapy reflects the valued role of newer agents in the management of myeloma patients. [Display omitted] [Display omitted] [Display omitted] DisclosuresAvivi:Tel Aviv Sourasky Medical center: Consultancy, Other: consultancy to :BMS Roche.

Prenatal cytogenetic diagnosis in Taiwan: a nationwide population-based study

Purpose: The goal of this study was to gain a better understanding of the status of advanced maternal age among criteria for provision of amniocentesis in pregnant women in Taiwan.Materials and method: Data of 315 670 second-trimester amniocenteses from 28 national certified cytogenetics laboratories were retrospectively analyzed from the Prenatal Genetic Diagnosis Declaring and Database System of the Health Promotion Administration, Ministry of Health and Welfare in Taiwan between 2006 and 2013.Results: The number of pregnant women undergoing amniocentesis in Taiwan between 2006 and 2013 increased, and the most common three indications for amniocentesis were advance maternal age (75.11%), abnormal second trimester maternal serum screening (13.22%) and abnormal sonographic finding (8.00%). Down syndrome was the most common autosomal abnormality identified (25.74%); Turner syndrome was the most common sex chromosome abnormality (7.04%). Of structural rearrangements, 26.93% were balanced translocations and 17.10% were unbalanced translocations. The greatest proportion of fetal chromosomal abnormalities was found in cases where parents were also affected (38.02%).Conclusions: Clinical workers should provide detailed genetic diagnostic information to pregnant women, especially those with the common amniocentesis indications, which will enable them to determine a birth plan.