TEST:

TruSight Myeloid Sequencing Panel

Six weeks later, while receiving the CD20-directed immunotherapies glofitamab and obinutuzumab, another bone marrow biopsy revealed eradication of the B-ALL and new involvement by pure erythroid leukemia (PEL). This patient's B-ALL lacked an IGH::BCL2 fusion resulting from t(14; 18), suggesting it did not arise through transformation of his prior FL/DLBCL. Instead, the B-ALL was likely caused by his extensive exposure to genotoxic chemotherapy drugs – including alkylating agents – for his preceding lymphomas. The shared cytogenetic and molecular features between this patient's B-ALL and PEL imply they are clonally related.

Only two patients in the MRD negative group received the FLT3 inhibitor midostaurin combined with chemotherapy...NGS-MRD was not prognostic in this cohort of FLT3-TKD mutated AML patients. Shared first authors: Isabell Arnhardt, Christian M Vonk Shared senior authorship: Michael Heuser, Peter J.M. Valk

Serial NGS-based studies in patients with TP53MT inform important clues to prognosis. While being a dominant or persistent TP53MT clone either at the same configuration or up-trending clonal burden post treatment equates to poor prognosis, it is crucial to study further those who display better clinical outcomes with standard-of-care treatment. Such information will be invaluable for making treatment decisions in this grave prognostic disease otherwise.

We analyzed 851 patients with MN at Karmanos Cancer Institute; 444 with serial samples. Among these patients, 174 (39%) had primary AML (pAML), 144 (32%) had secondary AML (sAML), 58 (13%) had MDS, 18 (4%) had MDS/MPN and 50 (11%) had MPN. Median age for primary AML was 56 ys (19-82), secondary AML was 64 ys (31-86) and MDS was 66 ys (23-90).

Here, we report a comprehensive analysis of baseline MCH mutational landscape in pts enrolled in the Fondazione Italiana Linfomi (FIL) MCL0208 phase 3 trial (NCT 02354313), evaluating lenalidomide maintenance vs observation after chemoimmunotherapy and ASCT in untreated MCL pts ≤ 65 years...In conclusion, we provided the M-CH mutational landscape at baseline in younger MCL pts and we showed for the first time the unfavorable clinical impact of large CH clones on MCL progression. Figure 1.

Aims: Here, we report a comprehensive analysis of baseline M-CHIP mutational landscape in pts enrolled in theFondazione Italiana Linfomi (FIL) MCL0208 phase 3 trial (NCT 02354313), evaluating lenalidomide (LEN)maintenance vs observation after chemoimmunotherapy and ASCT in untreated MCL pts ≤ 65 years... In conclusion, we provided the M-CHIP mutational landscape at baseline in younger MCL pts from aprospective clinical trial and we showed for the first time the unfavorable clinical impact of large CH clones onMCL progression. Further studies are ongoing on CH mutations detected during follow-up.

NPM1 mutations were identified in 21. 7% of 1,520 patients. Patients with NPM1mut were older and had higherWBC counts and LDH levels, and more often had a normal karyotype, but less adverse cytogenetic features,including monosomal karyotype and MR cytogenetic abnormalities at diagnosis compared to NPM1wtpatients.

Highly heterogeneous molecular genetic profile is present in patients from adverse risk group. Mutations ingenes that activate intracellular signaling pathways are most common in high-risk AML. The presence of morethan 6 mutations and ASXL1mut+ and SRSF2mut+ status negatively affect the survival of patients.

Notably, none of the patients received oral azacitidine as maintenance therapy... The results indicated that NPM1-/FLT3-ITD- and NPM1+/FLT3-ITD- subgroups in DNMT3A mutated AML benefit from allo-HSCT in CR1. Larger cohorts are warranted to validate these findings.

Notably, none of the patients received oral azacitidine as maintenance therapy... The results indicated that NPM1-/FLT3-ITD- and NPM1+/FLT3-ITD- subgroups in DNMT3A mutated AML benefit from allo-HSCT in CR1. Larger cohorts are warranted to validate these findings.

Hematologic malignancies developing in infancy and childhood - including ALL - have been described in two other patients with Weaver syndrome, although heightened cancer surveillance is not currently recommended. Interestingly, one of these patients - who harbored the same EZH2 E745K variant - developed non-Hodgkin lymphoma at 13 years old. This unusual case of ALL arising in both childhood and adulthood supports a potential link between Weaver syndrome and susceptibility to hematologic cancers.

It effectively allows for bypassing logistical, organizational, and financial burdens, as well as regulatory and ethical concerns. Ultimately, this enables explorative research inquiries into previously inaccessible data sets and offers the prospect of fully synthetic control arms in prospective clinical trials.

However, allo-HSCT failed to remedy the dismal outcomes for patients with MDS-biTP53. In conclusion, based on the evidence gathered from this cohort analysis, the 2022 WHO classification promote efficient segregation of this heterogeneous disease, with focus on the differences in molecular features and prognoses across different disease subtypes, leading to accurate MDS diagnosis and effective risk-adapted treatment.

The ASXL1 c.1934dupG variant is now routinely detected in myeloid NGS data using Oncomine Myeloid Assay GX v2 at VAF ³ 10%. Variant calls between 5% and 10% VAF are confirmed by orthogonal testing using the PCR sizing assay. The updated pipeline and sizing assay protocol are available to other laboratories.

Although somatic EZH2 mutations are not well described in B-ALL, germline EZH2 variants have been reported in Weaver syndrome: A rare autosomal dominant overgrowth disorder affecting 50 individuals to date. Close review of her medical record confirmed she had many clinical features of Weaver syndrome: Tall stature (5'11''), a distinctive facial appearance, intellectual disability, a history of developmental delay, and signs of pre- and postnatal overgrowth (99th percentile for childhood growth). Hematologic malignancies developing in infancy and childhood – including ALL – have been described in two patients with Weaver syndrome; however, heightened cancer surveillance is not currently recommended.

The rapid demise of these two patients with BRAF V600E-mutant AML highlights their poor prognosis. Few cases of BRAF V600E-mutant AML have been described in the literature, all of which reported poor survival. Intriguingly, most cases exhibited monocytic morphology and concurrent KMT2A rearrangements, as was observed in case 2.

We identified 3 cases of RUNX1-FPD from our institution with a range of clinical presentations, family histories, and molecular genetic findings. All RUNX1 variants were present with a VAF between 40% and 60%. Age of onset, prior history of bleeding disorder, prior thrombocytopenia, and family history were variable.

The Archer panel presented high performance and good coverage in GC-rich regions as CEBPA gene being a promising test in accuracy of somatic analysis and was validated in our laboratory.

The discovery of clonal hematopoiesis in this elderly woman with a wide spectrum of severe auto-inflammatory conditions supports a potentially intriguing link between somatic blood mutations and her adult- onset rheumatologic disorders. Mechanistically, disrupted SRSF2-mediated splicing may be driving aberrant antigen presentation by bone marrow-derived dendritic cells in her skin and other organs. Causal relationships between somatic blood mutations and autoimmune/auto-inflammatory conditions have been established in patients with hematologic malignancies and conditions like VEXAS syndrome.

Identification of an EZH2 p.E745K mutation about 50% VAF during tumor-only sequencing prompted careful review of the medical literature. Somatic EZH2 mutations are not well described in B-ALL, although germline EZH2 variants have been reported in Weaver syndrome, a rare autosomal dominant overgrowth disorder documented in 50 individuals to date. Close review of her medical record confirmed she had many clinical features of Weaver syndrome: tall stature (5'11''), a distinctive facial appearance, intellectual disability, a history of developmental delay, and signs of pre- and postnatal overgrowth (99th percentile for childhood growth).

We found that rare variants or mutations in myeloid neoplasm-associated genes were detected in PRCA. In our cohort, TET2, ASXL1, GATA2 were frequent in both primary and T-LGLL associated PRCA. Somatic mutation, Pure red cell aplasia, T cell leukemia

Our data show that the overall incidence of mutations increases by age up to 65 years, but thereafter remains stable, whereas certain genes have different age distributions. The strong impact of specific genetic alterations on survival in a large population-based study with real-life age distribution is shown. The clinical impact of combinations of specific mutations and karyotypes is being evaluated.

We observe that the compound genotype of ASXL1 mutation accompanied by a high mutant TET2 allele burden identified a distinct CMML population with significantly reduced survival, with independent prognostic relevance extrapolated to and validated in an independent cohort. Prospective and large-scale studies are warranted to support these observations and experimental studies are needed to ascertain the underlying mechanistic basis. Figure 1 TET2, Chronic myelomonocytic leukemia, ASXL1, Prognosis

IPSS-M improved prognostic discrimination and optimized treatments for patients with 2022 ICC-defined MDS. Patients with high, or very high-risk IPSS-M might benefit from HSCT. In addition to 2022 ICC and other parameters, IPSS-M also provided independent prognostication.

We are reporting a positive correlation between MYD88 allelic burden and Clonal B-cell fraction in LPL. Quantitative molecular assay for MYD88 might be useful for evaluation of residual disease in LPL.

mRNA expression analysis demonstrated that these mutations are strongly regulated by NMD machinery, leading to (almost) complete downregulation of the mutated WT1 allele. These results strongly question any direct effect of WT1 mutations in the leukemic blasts and therefore cannot explain the role of mutations in WT1 as a prognostic factor in AML.

This is the most comprehensive study assessing the impact of RUNX1mut in a large pediatric cohort. Unlike adult AML (Gaidzik et al., 2011 and 2016), our results provide evidence for a different clinicopathologic impact of RUNX1mut in pediatric AML, arguing against its classification as a distinct entity in childhood AML. These results may broaden the perspective on the pathological significance of RUNX1mut for leukemogenesis in pediatric AML.

Conclusion We provide evidences that VAF is critical for risk stratification in MDS patients and should be considered in novel scoring systems. Our data fostered our understanding of the mutation burden of the diseases and provided future patient-tailored therapeutic avenues.

FDA approved IDH2 inhibitor enasidenib in 2017 and IDH1 inhibitor ivosidenib in 2018 for adults with relapsed/refractory (R/R) IDH2 and IDH1MT AML. In May 2022, FDA approved ivosidenib combined with azacitidine for adults aged 75 years or older with newly diagnosed AML who cannot undergo intensive induction chemotherapy...IDHi-induced differentiation syndrome (DS) was reported in 6 patients (32%); 4 (21%) who developed DS received Olutasidenib, and 2 (11%) received Enasidenib...All patients with DS were treated with medication discontinuation and intravenous dexamethasone... Only a minority of patients with IDHMT AML had received IDHi-based therapy, which is attributed to patients diagnosed and treated before the FDA approval. Surprisingly, the median survival for patients who didn't receive IDHi therapy seemed not different, underscoring the other therapies' effects, including the influence of the venetoclax-based regimen. More extensive randomized studies are needed to compare the efficacy of IDHi to other therapies in treating IDHMT AML.

Figure 2. (A) Overall survival by IPSS-R, (B) Overall survival by IPSS-M.

The combination of the Illumina Trusight Myeloid Sequencing Panel and the Qiagen Clinical Insight Interpreter tertiary analysis software is a sensitive and specific approach for the detection and interpretation of clinically important variants in myeloid neoplasms.

The implementation of this NGS assay allows for a comprehensive evaluation of the mutational spectrum of myeloid malignancies in Lebanese patients. The detection of these actionable gene mutations contributes to an early diagnosis, appropriate individualized treatment, enhanced prediction of target therapy response, and improved clinical outcome.

Unsurprisingly, ASCT improved OS across all patients irrespective of induction type. Further study with a prospective randomised control trial is required.

Conclusion Patients that achieve CR/CRi until FU90 after DLIs have a significantly better outcome than the remaining patients. However, the MRD status in patients in CR/CRi at FU90 had no prognostic effect in the patient cohort investigated here.