Contiguous follicular lymphoma and follicular lymphoma in situ harboring N-glycosylated sites

International audienceFollicular lymphoma in situ (FLIS) is composed of a clonal B-cell population harboring the typical t(14;18) hallmark of follicular lymphoma (FL), forming unconventional BCL2 Bright CD10 + cell foci in an otherwise normal reactive lymph node (LN). The diagnosis of FLIS is made on the fortuitous discovery of unconventional BCL2 Bright CD10 + cell foci. 1 Several studies recently demonstrated that FLIS are already advanced precursors in follicular lymphomagene-sis, but not necessarily committed to malignant transformation. 2,3 However, the relationship between FLIS and FL still remains unclear, as only a minority (<5%) of FLIS patients eventually develop FL. This is in line with the usually indolent progression of the disease, and the genomic instability observed in FLIS cells, which can engage FL precursor cells either in an evolutionary malignant process, or to an evolutionary dead end. 4 We report the case of a 35-year old male patient who presented with a cervical adenopathy. Histological examination of the excised LN displayed an altered architecture suggestive of FL, consisting of high number of monomorphic large follicles, uniformly spread in the cortical and medullary areas. Most follicles contained a predominant population of small cleaved cells with scant macrophages and mitoses. The mantle zone was reduced or absent. However, in a minor cortical area, a few follicles showed features mimicking residual classical germ cells (GC), including a smaller size, higher cell polymorphism, and a preserved mantle zone (Figure 1A). The BCL2 immunostaining (clone 100) was negative in follicles displaying a typical FL pattern. In contrast, follicles located in the pseudo-residual area were BCL2bright, i.e. more strongly stained than the surrounding mantle zone and reactive T cells (Figure 1B). Most follicles were only slightly positive for Ki67 (Online Supplementary Figure S1A). Both BCL2 – and BCL2 + follicles were CD10 positive (Online Supplementary Figure S1B) and contained a BCL2/JH break-point evidenced by fluorescence in situ hybridization (FISH) (Figure 1C). Taken together these results suggested the diagnosis of simultaneous occurrence of BCL2 – FL (grade I/II) and of BCL2 + FLIS in the same LN. We decided to further analyze those two lesions independently, and performed macrodissection in order to proceed with individual molecular analyses when required. Sanger sequenc-ing revealed that both FLIS and FL shared the same BCL2/JH sequence at the t(14;18)+ breakpoint, and thus originated from the same clone (Figure 1D). We tested two other anti-BCL2 antibodies (E17, SP66) directed against other epitopes, but the staining remained BCL2-in the FL area of the LN, similar to the anti-BCL2 antibody (clone 100) staining (Figure 1E and F). We thus sequenced exons 1 to 3 of the BCL2 gene (B-cell CLL/lym-phoma 2, NG_009361.1). Punctual mutations, resulting in amino acid substitutions, were found in the FL component (Online Supplementary Table S1), and were indeed located in the targeted aa41 to aa54 epitope of clone 100 (mutation

Azacitidine Plus Venetoclax for the Treatment of Relapsed and Newly Diagnosed Acute Myeloid Leukemia Patients

Venetoclax (VEN) belongs the BH3-mimetic class that selectively targets BCL-2, activating apoptosis. The combination of VEN and azacitidine (AZA) has changed the paradigm of treatment of newly diagnosed (ND) acute myeloid leukemia (AML) patients ineligible for intensive chemotherapy. There is scarce evidence for the use of VEN&ndash;AZA for relapsed or refractory (R/R) AML. We compared the outcome of 39 R/R AML and 38 ND AML patients treated between 01/20 and 12/21. The median age was 69 (22&ndash;86) and 73 (61&ndash;81) in the R/R and ND groups, respectively. Adverse cytogenetics were found in 36% of patients in the R/R group and 59% of patients in the ND group. Overall response rate was 37% in R/R AML, including 13% CR, 8% CRi, 3% PR and 13% MLFS, and 58% in the ND AML, including 32% CR, 13% CRi and 13% MLFS. Adverse cytogenetics was associated with treatment failure in the R/R group (Relative Risk = 0.13, p = 0.005). Median overall survival (OS) was 5.9 months in the R/R group and 9.4 months in the ND group. Median OS was 2.2 months in the adverse cytogenetics group versus 8.7 months in the intermediate cytogenetics group in the R/R group (p = 0.02). Median leukemia-free survival was not different between the two groups (9.4 months and 10.3 months), indicating that VEN&ndash;AZA can be an efficient salvage treatment for selected R/R AML patients. In conclusion, VEN&ndash;AZA is a promising treatment for ND AML and for selected R/R AML patients

Clinical and molecular influences of H3K27me3 HIST1 mark in acute myeloid leukemia with normal cytogenetics

De nombreuses altérations épigénétiques ont été décrites dans les leucémies aiguës myéloïdes (LAM). Nous avons récemment mis en évidence un enrichissement anormal en la marque histone H3K27me3, située sur 70 kb du cluster HIST1 (6p22) dans 50% des échantillons de patients atteints d'une LAM avec un caryotype normal (CN). Nous étudions dans ce travail, les conséquences cliniques et moléculaires de cette marque. H3K27me3 HIST1high est associé à 1) une meilleure survie des patients en analyse multivariée 2) la sous-expression du mRNA de plusieurs gènes histones (HIST1H1D, HIST1H2BG et HIST1H2BH) 3) un enrichissement fonctionnel en gènes associés à la réponse immunitaire ou inflammatoire, en faveur d’un engagement dans la différenciation granulocytaire, surexprssion confirmée pour trois de ces gènes (CYBB, FCN1 et CLEC4A) par RT-qPCR dans les blastes de patients H3K27me3 HIST1high 4) une diminution de la quantité absolue de protéine histone linker H1d par spectormétrie de masse et par Western blot et 5) une meilleure sensibilité à la différenciation induite par l'acide rétinoïque dans la lignée OCI-AML3 avec un KD de H1d (augmentation de l’expression des marqueurs de différenciation CD11B et CD11C, présence de granules intra-cytoplasmiques et expression des gènes CYBB et ITGAM). En conclusion, le biomarqueur H3K27me3 HIST1high est associé à une meilleure survie dans les LAM-CN NPM1mut, un phénotype plus différencié des blastes et une sous-expression génique et protéique de certains histones incluant le sous-type histone linker H1d. H1d semble être important dans la différenciation des blastes de LAM NPM1mut et pourrait constituer une cible thérapeutique.The epigenetic machinery is frequently altered in acute myeloid leukemia (AML). We previously described an abnormal histone H3K27me3 repressive enrichment covering 70 kb on the HIST1 cluster (6.p22). In the present work, we further studied the medical significance and the molecular impact of this new epigenetic biomarker. We observed that H3K27me3 HIST1high is associated with 1) a better patients' outcome in multivariate analysis, 2) a lower histone mRNA expression of several histone genes (HIST1H1D, HIST1H2BG and HIST1H2BH), 3) a mature granulocytic gene expression profile including immune or inflammatory responsive genes, (we confirmed the higher expression of three of these genes, CYBB, FCN1 and CLEC4A, using qPCR in the H3K27me3 HIST1high patients' samples), 4) a decrease in histone linker H1d absolute protein abundance by Mass spectrometry and by Western blot analyses and 5) a better retinoic acid sensitization of the H1d KD OCI-AML3 cell line (i.e. increase of CD11B and CD11C expression on cell surface, higher percentage of cytoplasmic granules and mRNA up-expression of two mature granulocytic genes, CYBB and ITGAM). To conclude, this study showed that epigenetic silencing of the HIST1 locus by the H3K27me3 mark is associated with a better outcome, but also a mature gene expression profile in the NPM1mut subgroup of patients. We suggested that H1d has an important role of histone linker expression in AML blast cell differentiation. This protein could constitute a new epigenetic target

Venetoclax in Acute Myeloid Leukemia: Molecular Basis, Evidences for Preclinical and Clinical Efficacy and Strategies to Target Resistance

Venetoclax is a BH3-mimetics agent specifically interacting with the antiapoptotic protein BCL-2, facilitating cytochrome c release from mitochondria, subsequent caspases activation, and cell death. Utilization of venetoclax has profoundly changed the landscape of treatment for the poor-prognosis category of AML patients unfit for intensive chemotherapy. In the phase III VIALE-A study, Venetoclax, in combination with the hypomethylating agent azacitidine, showed a 65% overall response rate and 14.7-month overall survival, in comparison with 22% and 8 months in the control arm. These results led to the widespread use of venetoclax in this indication. Other combination regimens, consisting of low-intensity, intensive, or targeted therapies are currently under evaluation. Despite promising results, preventing relapses or resistance to venetoclax is still an unmet clinical need. Numerous studies have been conducted to identify and overcome venetoclax resistance in preclinical models or in clinical trials, including the inhibition of other antiapoptotic proteins, the induction of proapoptotic BH3-only proteins, and/or the targeting of the mitochondrial metabolism and machinery

Patients' Non-Medical Characteristics Contribute to Collective Medical Decision-Making at Multidisciplinary Oncological Team Meetings.

BACKGROUND:The contribution of patients' non-medical characteristics to individual physicians' decision-making has attracted considerable attention, but little information is available on this topic in the context of collective decision-making. Medical decision-making at cancer centres is currently carried out using a collective approach, at MultiDisciplinary Team (MDT) meetings. The aim of this study was to determine how patients' non-medical characteristics are presented at MDT meetings and how this information may affect the team's final medical decisions. DESIGN:Observations were conducted at a French Cancer Centre during MDT meetings at which non-standard cases involving some uncertainty were discussed from March to May 2014. Physicians' verbal statements and predefined contextual parameters were collected with a non-participant observational approach. Non numerical data collected in the form of open notes were then coded for quantitative analysis. Univariate and multivariate statistical analyses were performed. RESULTS:In the final sample of patients' records included and discussed (N = 290), non-medical characteristics were mentioned in 32.8% (n = 95) of the cases. These characteristics corresponded to demographics in 22.8% (n = 66) of the cases, psychological data in 11.7% (n = 34), and relational data in 6.2% (n = 18). The patient's age and his/her "likeability" were the most frequently mentioned characteristics. In 17.9% of the cases discussed, the final decision was deferred: this outcome was positively associated with the patients' non-medical characteristics and with uncertainty about the outcome of the therapeutic options available. LIMITATIONS:The design of the study made it difficult to draw definite cause-and-effect conclusions. CONCLUSION:The Social Representations approach suggests that patients' non-medical characteristics constitute a kind of tacit professional knowledge that may be frequently mobilised in physicians' everyday professional practice. The links observed between patients' attributes and the medical decisions made at these meetings show that these attributes should be taken into account in order to understand how medical decisions are reached in difficult situations of this kind

Erythroleukemia: Classification

Abstract Acute erythroid leukemia (AEL) is a rare (2%–5%) form of acute myeloid leukemia (AML). Molecular alterations found in AEL resemble those of other AMLs. We report a classification of AELs in three major classes, with different prognosis and some specific features such as a tendency to mutual exclusion of mutations in epigenetic regulators and signaling genes

Factors associated with final decisions made at MDT meetings: Univariate and multivariate comparisons (logistic regression).

<p>Factors associated with final decisions made at MDT meetings: Univariate and multivariate comparisons (logistic regression).</p

Distribution of patients’ non-medical characteristics mentioned at MDT meetings (N = 95 cases).

<p>Distribution of patients’ non-medical characteristics mentioned at MDT meetings (N = 95 cases).</p

Strategy of case-records selection for the analysis.

<p>Strategy of case-records selection for the analysis.</p

Factors associated with the mention of patients’ non-medical characteristics: univariate and multivariate comparisons (logistic regression).

<p>Factors associated with the mention of patients’ non-medical characteristics: univariate and multivariate comparisons (logistic regression).</p