Specific Chromosomal Abnormalities Research Articles

Embryo Mosaicism Rate in National Referral Hospital of Indonesia Detected Using Next-Generation Sequencing: A Retrospective Study.

Chromosomal mosaicism, a phenomenon observed in a minority of embryos, showcases its prevalence and inherent unpredictability, leading to variations in embryo mosaic rates across different centers. This research endeavors to assess the prevalence of mosaicism and its characteristics within the scope of our preimplantation genetic testing-A (PGT-A) services in Indonesia. Specifically focusing on our center's experience since 2020, this study aims to elucidate mosaic rates among embryos in our care. In a retrospective approach, we collected secondary data sourced from our PGT-A outcomes dating back to 2020. A total of 196 embryos underwent analysis, their characteristics were documented and presented descriptively. Notably, the incidence of specific chromosome abnormalities was outlined. We assess a comparative analysis to investigate the relationship between mosaicism and its corresponding clinical characteristics. In the analysis of 196 embryos, 106 (54.1%) displayed chromosomal anomalies spanning from low-level mosaicism to whole chromosome aneuploidy. Low mosaicism was observed in 25 (12.8%) of the embryos, while high mosaicism was identified in 8 (4.1%) embryos. Notably, low-level mosaicism predominated in chromosome 9 (n=10, 5.1%), whereas abnormality prevalence was highest in chromosome 21 (n=20, 10.2%). Statistical analysis revealed no significant disparity in mean maternal age among embryos with low-level mosaicism, high mosaicism, and normal chromosomes (33.88 vs. 35 vs. 33.26 years old, respectively). However, a statistically significant difference in mean maternal age (35.84 vs. 33.26 years) was observed between embryos with aneuploidy (monosomy or trisomy) and those with normal chromosomes. Furthermore, a significant difference in high mosaicism rates was detected in patients with unexplained infertility (P<0.05). In contrast to the study conducted elsewhere, our center had a higher mosaicism rate. Chromosomes 9, 8, and 6 were the most frequently affected. There was a significant difference in the high mosaicism rate for PGT-Arelated unexplained infertility causes.

HMGA2 overexpression with specific chromosomal abnormalities predominate in CALR and ASXL1 mutated myelofibrosis

HMGA2 overexpression with specific chromosomal abnormalities predominate in CALR and ASXL1 mutated myelofibrosis

Chromosomal Abnormalities in Chromosome 5 and Chromosome 8 in Iraqi Patients with Acute Myeloid Leukemia

Acute myeloid leukemia (AML) is the mostly diagnosed leukemia in adults, resulting from accumulate immature blasts in the bone marrow that are replaced instead of normal blasts that cannot function like normal blood cells. AML resulting from genetic abnormalities, including multiple gene mutations and chromosomal aberrations, are found in approximately 80% of children with AML. These shown correlations between specific recurrent chromosomal abnormalities and clinical-biological characteristics and outcomes, also, novel therapies are being developed that target some of the identified genetic defects. The aim of this study was to investigate and record the role of chromosome 5 and 8 aberrations in the development of newly diagnosed and relapsed cases of AML in Iraqi patients. Chromosomal changes were studied in peripheral blood samples from 30 Iraqi patient samples diagnosed with acute myeloid leukemia and divided into 9 newly diagnosed and 13 chemotherapy-treated subjects who were incomplete remission and 8 relapsed subjects. The results of the chromosomal analysis of this study revealed numerical and structural abnormalities of both chromosomes: 5 and 8 in 16 (53.33%) and 12 (40%), respectively, within a complex karyotype and high frequencies in numerical abnormalities of chromosomes for chromosome 5 and structural abnormalities for chromosome 8. Chromosomal aberrations involving chromosomes 5 and 8 are linked to AML development and prognosis, and their presence in a complex karyotype can lead to disease progression and the worst prognosis, especially for abnormalities on chromosome 5, which were detected the most frequently. Future molecular genetic tests and complete karyotype analysis should enhance AML diagnosis and treatment.

Approach and Management of Pregnancies with Risk Identified by Non-Invasive Prenatal Testing.

This study represents our second investigation into NIPT, involving a more extensive patient cohort with a specific emphasis on the high-risk group. The high-risk group was subsequently divided into two further groups to compare confirmed cases versus unconfirmed via direct methods. The methodology encompassed the analysis of 1400 consecutive cases from a single genetic center in western Romania, where NIPT was used to assess the risk of specific fetal chromosomal abnormalities. All high-risk cases underwent validation through direct analysis of fetal cells obtained via invasive methods, including chorionic villus sampling and amniocentesis. The confirmation process utilized QF-PCR, karyotyping, and SNP-Array methods customized to each case. Results: A high risk of aneuploidy at NIPT was identified in 36 out of 1400 (2.57%) cases and confirmed in 28 cases. The study also detected an increased risk for copy number variations (CNVs) in 1% of cases, confirmed in two instances involving one large microdeletion and one large microduplication. Trisomy 21 was the exclusive anomaly where NIPT confirmed all cases with identified risk. High-risk NIPT results which were not validated by invasive methods, were classified as false positives; parents in these cases determined to continue the pregnancy. In conclusion, NIPT can serve as a screening method for all pregnancies; however, in high-risk cases, an invasive confirmation test is strongly recommended.

MicroRNA dysregulation in myelodysplastic syndromes: implications for diagnosis, prognosis, and therapeutic response.

Myelodysplastic syndromes (MDS) are a group of malignant clonal hematological disorders with heterogeneous clinical course and risk of transformation to acute myeloid leukemia. Genetic and epigenetic dysregulation, including alterations in microRNA (miRNA) expression, plays a pivotal role in MDS pathogenesis influencing disease development and progression. MiRNAs, known for their regulatory roles in gene expression, have emerged as promising biomarkers in various malignant diseases. This review aims to explore the diagnostic and prognostic roles of miRNAs in MDS. We discuss research efforts aimed at understanding the clinical utility of miRNAs in MDS management. MiRNA dysregulation is linked to specific chromosomal abnormalities in MDS, providing insights into the molecular landscape of the disease. Circulating miRNAs in plasma offer a less invasive avenue for diagnostic and prognostic assessment, with distinct miRNA profiles identified in MDS patients. Additionally, we discuss investigations concerning the role of miRNAs as markers for treatment response to hypomethylating and immunomodulating agents, which could lead to improved treatment decision-making and monitoring. Despite significant progress, further research in larger patient cohorts is needed to fully elucidate the role of miRNAs in MDS pathogenesis and refine personalized approaches to patient care.

A cytogenetic Study of Down’s Syndrome in Iraqi Population

Down’s syndrome is the most common type of chromosomal abnormality found in neonates. It is associated with characteristic abnormal facial features and mental retardation. However, clinical diagnosis is not sufficient to confirm the disease. Classical karyotyping has long been used as diagnostic tool for confirming Down’s syndrome. The present study was conducted to confirm the cytogenetic composition of individuals suspected with Down’s syndrome after the preliminary clinical diagnosis in Basra, Iraq. Karyotype analysis was carried out for 249 suspected cases of Down’s syndrome. 249 patients were confirmed to have Down’s syndrome after cytogenetic tests. Nearly 90% of these patients had pure trisomy 21 and about 5% of cases had translocations. Less than 4% of cases were reported having mosaic trisomy and only 1.4% patients carried additional chromosomal abnormalities. In our study, the disease showed significant gender bias, with an excess of males than females (sex ratio was 1.39). The identification of specific chromosomal abnormalities in different variants of Down’s syndrome patients work as first step towards improving their quality of life. The study also provides the basic information required to forecast the various complications associated with Down’s syndrome.

Role of Interphase FISH Assay on Air-Dried Smears in Identifying Specific Structural Chromosomal Abnormalities among Pediatric Patients with Acute Leukemias.

The online version contains supplementary material available at 10.1007/s12288-023-01699-2.

Prevalence of Bruxism in Down Syndrome Patients: A Systematic Review and Metanalysis.

Bruxism is a parafunctional activity characterized by grinding or clenching of teeth and is a common oral health concern in individuals with Down syndrome (DS). Understanding the prevalence of bruxism in this population is crucial for developing effective management strategies. This systematic review and meta-analysis is aimed to investigate the prevalence of bruxism among individuals with DS and explore its association with other oral health issues; METHODS: A comprehensive search was conducted across multiple electronic databases to identify relevant studies. Cross-sectional and observational studies were included. Data on bruxism prevalence and associated factors were extracted, and a meta-analysis was performed using both fixed and random-effects models of MedCalc software. Heterogeneity among studies was assessed using I2 statistics. New Castle - Ottawa scale was used to evaluate methodological quality of the included studies RESULTS: Eight studies met the predefined inclusion criteria and were included in the analysis. Seven studies used a questionnaire to assess bruxism. The pooled proportion estimate for occurrence of Down's syndrome across the included studies was found to be 0.33 (95% CI: 0.22-0.45) as per the RE model and 0.35(95% CI: 0.31-0.450) as per FE model in the quantitative analysis. All studies exhibited good methodological quality CONCLUSION: This systematic review and meta-analysis provide evidence of a significant prevalence of bruxism among individuals with Down syndrome. The findings highlight the association of bruxism with other oral health issues and specific chromosomal abnormalities. Comprehensive oral health assessments, including diagnostic procedures like PSG, are essential for addressing the unique oral health needs of individuals with DS. Further studies are recommended with a valid tool for the diagnosis. Early interventions and management strategies need to be tailored to this population, considering the multifaceted nature of oral health concerns in individuals with DS.

Genomic landscape of TP53-mutated myeloid malignancies

AbstractTP53-mutated myeloid malignancies are associated with complex cytogenetics and extensive structural variants, which complicates detailed genomic analysis by conventional clinical techniques. We performed whole-genome sequencing (WGS) of 42 acute myeloid leukemia (AML)/myelodysplastic syndromes (MDS) cases with paired normal tissue to better characterize the genomic landscape of TP53-mutated AML/MDS. WGS accurately determines TP53 allele status, a key prognostic factor, resulting in the reclassification of 12% of cases from monoallelic to multihit. Although aneuploidy and chromothripsis are shared with most TP53-mutated cancers, the specific chromosome abnormalities are distinct to each cancer type, suggesting a dependence on the tissue of origin. ETV6 expression is reduced in nearly all cases of TP53-mutated AML/MDS, either through gene deletion or presumed epigenetic silencing. Within the AML cohort, mutations of NF1 are highly enriched, with deletions of 1 copy of NF1 present in 45% of cases and biallelic mutations in 17%. Telomere content is increased in TP53-mutated AMLs compared with other AML subtypes, and abnormal telomeric sequences were detected in the interstitial regions of chromosomes. These data highlight the unique features of TP53-mutated myeloid malignancies, including the high frequency of chromothripsis and structural variation, the frequent involvement of unique genes (including NF1 and ETV6) as cooperating events, and evidence for altered telomere maintenance.

P-735 Are there any differences in preimplantation blastocyst chromosomal abnormalities between polycystic ovary syndrome patients and controls? A multi-center retrospective cohort study

Abstract Study question Are there possible differences in the preimplantation blastocyst chromosome aberrations between polycystic ovary syndrome (PCOS) patients and controls? Summary answer PCOS lowers aneuploidy risk but increases mosaicism risk in preimplantation embryos, and PCOS-related insulin resistance should be investigated as a potential cause. What is known already PCOS is thought to alter granulosa cell functions and effects on oocyte development, potentially leading to chromosomal abnormalities and increasing the risk of pregnancy loss. However, few studies have compared the chromosomal status of preimplantation embryos from PCOS patients to those from healthy controls. Study design, size, duration This was a multi-center retrospective cohort study conducted at 3 IVF centers. A total of 707 blastocysts from 147 PCOS patients and 3006 blastocysts from 821 control women receiving preimplantation genetic testing (PGT) between 2015 and 2021. Participants/materials, setting, methods PGT was performed using next-generation sequencing (NGS). Differences in the chromosome aberrations were compared. Multivariate and subgroup analyses were conducted to control for confounders. Influences of insulin resistance and PCOS phenotype on the risks of specific chromosomal abnormalities were also examined. Main results and the role of chance Compared to controls, blastocysts from PCOS patients demonstrated lower aneuploidy rate (14.7% vs. 25.4%, P &amp;lt; 0.001) but greater mosaicism rate (16.5% vs. 8.7%, P &amp;lt; 0.001). Multivariable analysis adjusting for age, IVF center, body mass index (BMI), total gonadotropin (Gn) dose, and peak serum estradiol (E2) on trigger day, identified PCOS as an independent protective factor against embryonic aneuploidy (adjusted OR = 0.56, 95% CI, 0.42–0.76, P&amp;lt;0.001) but a risk factor for embryonic mosaicism (adjusted OR = 2.05, 95% CI 1.48–2.85, P&amp;lt;0.001). These differences in chromosomal spectrum were also observed in subgroups stratified by age, IVF center and BMI. Further multivariate analysis suggested that insulin resistance could be responsible for the increased risk of embryonic mosaicism among PCOS patients (OR = 1.89, 95% CI, 1.06–3.34, P=0.03). Limitations, reasons for caution The main limitation is the retrospective design. Besides, methodological uncertainties still exist in detecting mosaicism in current reproductive medicine. A longer-term, multiple-center prospective study with greater methodological standardization is warranted to confirm the present results. Wider implications of the findings These results suggest that PCOS pathology may increase the risk of mitosis errors but not the risk of meiosis errors during oocyte development, and PCOS-related insulin resistance should be investigated as a potential cause. Trial registration number 2020[325]

Features of chromosomal abnormalities in relation to consanguinity: analysis of 10,556 blastocysts from IVF/ICSI cycles with PGT-A from consanguineous and non-consanguineous couples

Consanguineous marriage is defined as marriage between first or second-degree cousins, with high prevalence in many cultures and societies. Descendants from consanguineous unions have an increased risk for genetic diseases. Additionally, in consanguineous couples, chromosomal disjunction during embryogenesis could also be affected, increasing the risk of chromosomal errors. Nowadays, genomic testing allows to identify new genetic syndromes and variants related to copy-number variations (CNV), including whole chromosome, segmental and micro-segmental errors. This is the first study evaluating chromosomal ploidy status on blastocysts formed from consanguineous couples during IVF/ICSI treatments with Preimplantation Genetic Testing for Aneuploidies (PGT-A), compared to non-consanguineous couples. Although consanguine couples were significantly younger, no differences were observed between groups for fertilisation rate, blastulation rate and euploidy rate, once adjusted by age. Nevertheless, the number of blastocysts biopsied on day 5 was lower for consanguine couples. Segmental errors, and aneuploidies of chromosomes 13 and 14 were the most prominent abnormalities in relation to consanguinity, together with errors in chromosome 16 and sex chromosomes when the female partner was younger than 35. Once euploid blastocysts were considered for subsequent frozen embryo transfer, pregnancy outcomes were similar in both groups. The current findings point toward the fact that in consanguine unions, not only the risk of having a child with genetic disorders is increased, but also the risk of specific chromosomal abnormalities seems to be increased. Premarital counselling and tailored reproductive treatments should be offered to these couples.

Isolated aberrant right subclavian artery: an underlying clue for genetic anomalies

Objective Aberrant right subclavian artery (ARSA) is known to be associated with specific chromosomal abnormalities. However, there is no agreement regarding clinical decisions related to isolated ARSA. This study evaluated the association between ARSA and genetic abnormalities to provide evidence for prenatal consultation and the postpartum management of isolated ARSA. Methods This single-center cross-sectional study involved fetuses diagnosed with ARSA between January 2014 and May 2021. A range of data was recorded for each patient, including screening ultrasound, fetal echocardiograms, genetic results, postnatal information, and follow-up records. Results ARSA was detected in 151 fetuses, of which 136 were considered isolated cases. The remaining 9.9% (15/151) of cases had cardiac and/or extracardiac abnormalities or soft markers. Data from karyotype analysis and chromosomal microarray analysis (CMA) were available for 56 and 33 (out of 56) fetuses, respectively. Genetic abnormalities were detected in 10.7% of fetuses (6/56). Of these, 4.4% (2/45) and 36.4% (4/11) were associated with isolated and non-isolated ARSA, respectively, with a significant difference between these two groups regarding the frequency of genetic abnormality (p = 0.011). The analysis detected Klinefelter Syndrome (47, XXY) and 16p11.2 microdeletion in two isolated cases. One case each of trisomy 21 and 22q11.2 deletion, and another case of 47, XXY, were detected in fetuses with cardiac anomalies. Partial 5q deletion was found in a fetus with extracardiac malformations. In total, 141 of the fetuses survived after birth; termination of pregnancy was performed for 10 fetuses; only two fetuses had mild symptoms of dysphagia. Conclusion ARSA may be an underlying ultrasonic clue for genetic anomalies even in isolated ARSA. Fetuses with isolated ARSA cannot be ruled out for invasive antenatal diagnosis.

Evaluation of the cytogenetic profile in patients with acute leukaemia.

Acute leukaemia (AL) is aheterogeneous neoplastic disease that occurs by thegrowth ofabnormal lymphoid and myeloid cells in thebone marrow and blood leading to acute myeloid leukaemia (AML) and acute lymphocytic leukaemia (ALL). Conventional cytogenetics is acharacteristic technique to hunch chromosomal abnormalities, it helps in thediagnosis and therapeutic approach ofthedisease by themolecular cytogenetics technique offluorescence in situ hybridization (FISH). Chromosomal abnormalities in AL are performed by karyotyping to confirm specific chromosomal abnormalities using FISH. Thedescriptive study included 42 clinically diagnosed AL patients. Karyotyping analysis was performed using thestandard Giemsa banding procedure. To confirm specific chromosomal abnormalities and all culture failure (CF) cases, FISH was done. Among 42 cases, 29 (69.4%) males and 13 (30.9%) females, AML comprised 22 (52.38%) cases, ALL 14 (33.33%) cases, and AL 6 (14.2%) cases. Normal karyotype was found in 18 (42.85%), abnormal karyotype in 16 (39.09%), and 8 (19.09%) were CF. Specific abnormalities oft(15;17), hyperdiploidy; t(3;3) with monosomy 7 in; del(9q22); del(2p); del(17p); del(Xq); 1~2 dmin; der(3); +11, +13 and composite karyotype. Hypodiploidy was strongly associated with AL, which signifies theloss ofchromosomes causing potential risk. Composite karyotype, rare t(3;3) double minutes, +11,+13, del(9q), and del(Xq) were thenovel findings reported in theSouth Canara region ofKarnataka. Despite other molecular techniques, conventional cytogenetics remains thebaseline in thediagnosis ofmalignancies.

O-114 Preimplantation genetic testing for monogenic diseases without necessary familial members

Abstract Preimplantation genetic testing for monogenic diseases (PGT-M) is available for any single gene defects theoretically, as long as the disease-causing locus has been unequivocally identified. However, PGT-M has faced great challenges for couples with more than one genetic mutation, de novo mutations or without essential family members for linkage analysis. In order to overcome these challenges, we developed a comprehensive strategy which includes preimplantation genetic testing for aneuploidies (PGT-A) and chromosomal structural rearrangements (PGT-SR) study, as well as monogenic diagnosis study for couples bearing de novo mutations or without necessary familial members. The innovation of our strategy is to use the gamete (polar body or single sperm) or embryo as proband for linkage analyses (GEPLA) to detect an embryo’s carrier status. GEPLA strategy uses affected embryo as proband or gametes carrying mutant alleles as proband for SNP linkage analyses, that were suitable for all types of patients mentioned above. Using this novel developed method, nine autosomal dominant polycystic kidney disease (ADPKD) couples with either de novo mutation or without a positive family history were recruited. Moreover, other two couples with single gene disorders (SMA and NF2) which lack of essential family members for linkage analysis including de novo mutation, and also combined with reciprocal translocation were recruited too. For nine ADPKD couples, a total of 34 embryos from 13 PGT-M cycles were examined and two couples were successfully delivered healthy babies. For the other two couples, 15 embryos were screened, and two embryos were determined as free of the monogenic disease and specific chromosomal abnormalities created by reciprocal translocations. This study provides a creative approach for embryo diagnosis of patients with de novo mutations or patients lacking of essential family members for linkage analysis, and even for reciprocal chromosome translocation concurrently.

Genetic Counseling and Management: The First Study to Report NIPT Findings in a Romanian Population

Background and Objectives: Non-invasive prenatal testing (NIPT) has been confirmed as the most accurate screening test for trisomies 21, 18, 13, sex chromosomes aneuploidies and several microdeletions. This study aimed to assess the accuracy of cell free DNA testing based on low-level whole-genome sequencing to screen for these chromosomal abnormalities and to evaluate the clinical performance of NIPT. Materials and Methods: 380 consecutive cases from a single genetic center, from Western Romania were included in this retrospective study. Cell-free nucleic acid extraction from maternal blood, DNA sequencing and analysis of sequenced regions were performed by BGI Hong Kong and Invitae USA to determine the risk of specific fetal chromosomal abnormalities. In high-risk cases the results were checked by direct analysis of fetal cells obtained by invasive methods: 6 chorionic villus sampling and 10 amniocenteses followed by combinations of QF-PCR, karyotyping and aCGH. Results: NIPT results indicated low risk in 95.76% of cases and high risk in 4.23%. Seven aneuploidies and one microdeletion were confirmed, the other results were found to be a false-positive. A gestational age of up to 22 weeks had no influence on fetal fraction. There were no significant differences in fetal fraction across the high and low risk groups. Conclusions: This is the first study in Romania to report the NIPT results. The confirmation rate was higher for autosomal aneuploidies compared to sex chromosome aneuploidies and microdeletions. All cases at risk for trisomy 21 were confirmed. Only one large fetal microdeletion detected by NIPT has been confirmed. False positive NIPT results, not confirmed by invasive methods, led to the decision to continue the pregnancy. The main limitation of the study is the small number of patients included. NIPT can be used as a screening method for all pregnancies, but in high-risk cases, an invasive confirmation test was performed.

Secondary myelodysplastic syndromes identified via next-generation sequencing in a non-small cell lung cancer patient

BackgroundMyelodysplastic syndrome (MDS) is a group of clonal disorders characterized by ineffective and dysplastic hematopoiesis in the bone marrow with a high risk of progression to leukemia. Many studies have demonstrated that chemo-radiotherapy for cancer patients and exposure to certain chemicals may increase the risk of secondary MDS, which is characterized by specific chromosomal abnormalities and genomic alterations. Since next-generation sequencing (NGS) has been widely used for the diagnosis of cancer patients, advanced analysis of the sequencing data may provide supplementary information for secondary MDS.Case presentationA male patient with non-small cell lung cancer (NSCLC) and bone metastases has presented distal obstructive inflammation, the enlargement of the left hilar, mediastinal lymph node metastases, and multiple bone metastases. This patient has undergone long-term exposures to certain chemicals. Moreover, the deletion of chromosome 7 and 5q is detected in his peripheral blood sequencing, indicating secondary MDS, subsequently confirmed by bone marrow examination.ConclusionIn this case, an NSCLC patient was diagnosed with secondary MDS via NGS analysis, indicating that the NGS analysis may serve as supplementary for diagnosis of secondary MDS and provide useful information of therapeutic regimens for subsequent-line treatment of EGFR-mutated lung cancer. To the best of our knowledge, this is the first report of acquired MDS in a lung adenocarcinoma patient.

NEUROBLASTOMA IN A CASE OF CONGENITAL ADRENAL HYPERPLASIA

The majority of neuroblastomas are sporadic and not correlated with any specific constitutional germline chromosomal abnormality, inherited predisposition, or associated congenital anomalies. We report here a 1.5-year-old girl with a diagnosis of 21 hydroxylase deficiency and neuroblastoma. Neuroblastoma in a known case of congenital adrenal hyperplasia has rarely been reported. Based on our literature review, this is the fifth case report of congenital adrenal hyperplasia and neuroblastoma.

P-161: Ultra-high-risk prognostic significance of double and triple hit myeloma

<h3>Background</h3> Association of any of 2 or ≥3 specific high-risk chromosomal abnormalities (HR-CA: t(4;14), t(14;16), t(14;20) and +1q21) in multiple myeloma (MM) characterizes double or triple hit MM. <h3>Aims</h3> The aim of study was to evaluate prognostic significance of iFISH findings of double and triple hit MM on the course of disease. <h3>Methods</h3> The study analysed iFISH findings, clinical and laboratory characteristics of 502 newly diagnosed, transplant ineligible patients (pts, 259 male; 243 female, mean age 62yrs, range 35-85yrs) during period 2009-2021. According to the MM type, IgG MM was found in 297pts (60.4%), IgA in 100 (20.3%), BJ in 88 (17.9%), IgD in 5pts (1.0%) and IgM and non-secretory disease was found at 1pts each (0.2%). The clinical stage (CS, Durie-Salmon) III was confirmed in 397pts (79.6%); II 73pts (14.6%); and I with symptomatic disease 29pts (5.8%). The ISS score 1 had 135pts (28.7%), ISS2 112pts (23.8%), and 224pts (47.6%) had ISS3. Revised ISS score 1 (R-ISS1) was found in 100pts (21.8 %); R-ISS2 in 254pts (55.5%), and R-ISS3 in 104pts (22.7%). Renal impairment had 160pts (32%). Treatment with thalidomide (Thal) based chemotherapy (HT) was applied in 254pts (50.7%); bortezomib (Bz) based HT in 186pts (37.1%), while 56pts (11.2%) were treated with conventional HT. <h3>Results</h3> Finding of double and triple hit MM was identified in 21pts (6.8%): 17pts with double; 4pts with triple hit MM (12 male, 9 female, mean age 65yrs, range 44-81yrs). The IgG MM was present in 11pts, IgA in 5pts, and BJ in 5pt. Majority, 17pts were in III CS, and 4pts in II CS. The ISS3 score had 14pts, ISS2 5pts, ISS1 2pts, while R-ISS3 was found in 12pts, and R-ISS2 in 9pts. The overall treatment response (ORR, ≥PR) was significantly lower in pts with double and triple hit MM in comparison to the other MM patients (61.9% vs. 78.2%, p=0.018). The average PFS of patients with double and triple hit MM was significantly shorter in comparison to other MM patients, (17.0m vs. 48.0m; Log Rank=12.471, p<0.01); as well as the average OS of patients with double and triple hit MM (18.0m vs. 64.0m; Log Rank=10.593, p<0.01). Univariate Cox regression analysis pointed out the age (p<0.01), elevated LDH (p<0.01), R-ISS score (p<0.01) and presence of double and triple hit MM (p<0.01), as the major prognostic factors with impact on the OS of myeloma patients. Furthermore, the multivariate analysis confirmed major prognostic significance of age (p=0.02) and double and triple hit myeloma (p<0.01) for the OS. <h3>Conclusion</h3> Presence of double and triple hit myeloma indicates ultra high-risk course of disease, consequently indicating necessity of intensive treatment approach with new treatment modalities.

O-085 In-depth analysis of embryo development: Differences among monosomic, trisomic and chromosomally chaotic embryos compared to euploid embryos

Abstract Study question Is there any visible variation in the development of aneuploid embryos depending on the type of chromosome abnormality? Summary answer There were significant visible differences in the development of euploid, monosomic, trisomic and, especially, chaotic embryos. What is known already Aneuploidy rates are remarkably high in in vitro fertilized human embryos, with up to 50% of embryos diagnosed as aneuploid based on preimplantation genetic testing for aneuploidies (PGT-A). However, very little is known about the impact of specific aneuploidies during the early human embryo development. A recent publication showed that embryos with single chromosomal gain or loss reached the blastocyst stage later or earlier depending on the chromosome affected (Shahbazi et al., 2020). In our study, we wanted to detect observable differences in embryo behavior between embryos with different chromosomal abnormalities during the entire in vitro development. Study design, size, duration This was a retrospective study including 2,500 blastocysts with PGT-A results. Embryos were cultured in EmbryoScope systems until the fifth/sixth day of development (up to the time of trophectoderm biopsy). Automatic-annotations for division times and quality gradings were supervised routinely by senior embryologists using Guided Annotations Tool. Out of the total, 1,000 were euploid embryos used for reference and 1,500 were aneuploid embryos with one or more defects, including monosomic, trisomic and chromosomally chaotic embryos. Participants/materials, setting, methods Chromosome analysis was performed using next-generation sequence technology. Then, an in-depth analysis of time-lapse videos and supervised-automatic annotations was performed. We calculated the proportion of embryos, in each aneuploid category, that reached one specific event later than the expected value for euploid embryos plus one standard deviation. Later, we calculated the “relative risk” of an embryo of reaching the milestone late. We did the same for the time between milestones and for pairs of milestones. Main results and the role of chance Every aneuploid category was more likely to reach each specific embryo developmental event later than euploid embryos and the time gaps between developmental milestones were also statistically longer in aneuploid embryos (p &amp;lt; 0.0001). The following results were the most interesting relative risks (RR) when we compared the proportion of embryos (in each aneuploid category) to the proportion of euploid embryos (RR for euploid = 1). For reaching the division time to two cells (t2): 1.31 in monosomic embryos, 1.50 in trisomic embryos and 2.43 in chaotic embryos. For the division time to four cells (t4): 1.42 in monosomic embryos, 1.54 in trisomic embryos and 3.07 in chaotic embryos. For the division time to eight cells (t8) and the time of starting blastulation: 1.45 in monosomic embryos, 1.22 in trisomic embryos and 2.74 in chaotic embryos. Combined milestones were stronger indicators than each milestone by itself, the RR were: 1.63 in monosomic embryos, 1.81 in trisomic embryos and 3.35 in chaotic embryos for t2 and t4; 1.50 in monosomic embryos, 1.80 in trisomic embryos and 2.84 in chaotic embryos for t2 and t8; 1.46 in monosomic embryos, 1.90 in trisomic embryos and 3.43 in chaotic embryos for t4 and t8. Limitations, reasons for caution At this stage, we did not go down to specific chromosome abnormality as there were very few cases in each fully detailed category. Also, not all the embryos reached every developmental milestone. Wider implications of the findings Aneuploid embryos were significantly different from euploid embryos in the first five days of development. A large proportion of aneuploid embryos could be rejected because their developmental milestones falling outside the normal range. This could form part of an automated system for determining euploidy/aneuploidy from observation of embryos in vitro. Trial registration number 1902-VLC-018-MM

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

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