bluuebunny/crossref_metadata_2025 huggingface dataset:
An Accurate Prognostic Index Model for Adult HLH (HHLWG-HPI): A Multicenter Retrospective Analysis in China
Authors: Shen Ziyuan , Jin Yingliang , Sun Qian , Xu Linyan , Qiao Jianlin , Huang Shuiping , Xu Kailin , Sang Wei
Published: November 2021

Hemophagocytic lymphohistiocytosis (HLH) is an immune disorder with rapid progression and poor survival. Individual treatment strategy is restricted, due to the absence of precise stratification criteria. In this multicenter retrospective study, we aimed to develop a feasible prognostic model for adult HLH in China. A total of 270 newly diagnosed patients of adult HLH were retrieved from Huaihai Lymphoma Working Group (HHLWG), of whom 184 from 5 medical centers served as derivation cohort, and 86 cases from 3 other centers served as validation cohort. X-Tile program and Maxstat analysis were used to identify optimal cut-off points of continuous variables; univariate and multivariate Cox analysis were used for variables selection, and Kaplan-Meier curve was used to analyze the value of variables on prognosis. The C-index, brier score and calibration curve were used for model validation. Multivariate analysis showed that age, creatinine, albumin, platelet, lymphocyte ratio and alanine aminotransferase were independent prognostic factors. By rounding up the hazard ratios from 6 significant variables, a maximum of 9 points was assigned. The final scoring model of HHLWG-HPI was identified with four risk groups: low risk (≤3 pts), low-intermediate risk (4 pts), high-intermediate risk (5-6 pts), and high risk (≥7 pts), with 5-year overall survival rates of 68.5%, 35.2%, 21.3%, and 10.8%, respectively. The C-indexes were 0.796 and 0.758 in derivation and validation cohort by using bootstrap resampling program. In conclusion, the HHLWG-HPI model provides a feasible and accurate stratification system for individualized treatment strategy in adult HLH.

Disclosures

No relevant conflicts of interest to declare.
Epidemiology and Risk Factors for Infections in Myelodysplastic Syndromes
Authors: Poloni Antonella , Carturan Alberto , Pianelli Michelangelo , Morsia Erika , Lame Dorela , Olivieri Attilio
Published: November 2021

Background: Patients with MDS are at high risk of developing infections, which still represent the most frequent complications and the main cause of mortality and morbidity. We conducted a retrospective study with the aim of evaluating incidence, characteristics and outcome of infectious events in patients with myelodysplastic syndromes (MDS) and the risk factors associated with them.

Patients and methods: 200 patients with MDS or LMMC followed by the Ancona Hematology Clinic in the last 15 years were included. In this group, we evaluated severe infectious episodes and associated risk factors, relating to patient, disease and therapy.

Results: In a cohort of 200 patients, we detected 65 cases of infections of higher than third grade according to WHO. Pneumonia is the most frequent infection, especially on a bacterial basis. Risk factors that have shown significant association with infections ≥ G3 are: comorbidity (p = 0.0086), hemoglobin level (P = 0.0111), neutropenia (P = 0.0152), transfusion dependence during the course of disease (P <0.0001) and hypomethylating therapy (P = 0.0014). A subanalysis confirmed comorbidities as risk factors for the development of pneumonia (P = 0.0322) and azacitidine therapy as a risk factor for sepsis (P = 0.0038). The overall survival of the cohort is approximately 7.5 months and the predictive factors of survival are patient age (P = 0.0005), WHO disease class (P = 0.001), R-IPSS (P <0.0001) and transfusion-dependence during the course of the disease (P <0.0001). Multivariate analysis confirmed age (P = 0.0002), R-IPSS (P = 0.0043) and transfusion-dependence (P = 0.0006) as predictors of survival. Infections above grade 3 do not impact survival. Otherwise, there was a trend of reduced survival in patients who had pneumonia (P = 0.0612) or sepsis (P = 0.0927).

Conclusion: Patients with MDS have a high probability of encountering an infectious problem in the course of the natural history of the disease. In one out of five cases, there are more than one infectious event. Particularly at risk of infection are patients with comorbidities, neutropenia (PMN <800 / mm³), significant transfusion needs and on therapy with hypomethylating agents. These patients should be carefully monitored for infectious complications.

Disclosures

No relevant conflicts of interest to declare.
Eltrombopag As Initial Monotherapy for Transfusion Dependent Patients with Low to Intermediate Risk-1 Myelodysplastic Syndrome: Single Center Experience
Authors: Shen Yingying , Li Hangchao , Xiang Jingjing , Zhu Ni , Li Yuzhu , Liu Qi , Dong Huijie , Wang Bo , Ye Baodong , Lin Shengyun , Shen Yiping , Wu Dijiong
Published: November 2021

Myelodysplastic syndrome (MDS) is a group of disorders involving hematopoietic dysfunction, with a leukemic transformation characteristic. The risk based therapeutic strategy was well applied in clinical. And for patients with IPSS low or intermediate risk-1, NCCN guideline recommended hematopoietic stimulation therapy, immune suppression therapy (IST), as well as demethylation therapy, along with best supportive care. Eltrombopag (EPAG) was firstly approved as the second line option for immune thrombocytopenia (ITP), and recently it was also investigated for treatment of aplastic anemia and MDS. In MDS, based on available reports, the EPAG was mostly used along with hypomethylating-agent, or as salvage treatment after treatment failure. However, there was study showed that the combination of EPAG to azacitidine (AZA) may worsen platelet recovery, with lower response rate and increased trend of leukemic transformation. The successful EPAG monotherapy for MDS case was report, with complete hematologic remission and blasts elimination.

To date, limited study was focus on investigating the benefit of EPAG as initial monotherapy for transfusion dependent (TD) patients with low to intermediate risk-1 MDS, as well as its safety. Herein, we reviewed cases of elderly TD-MDS patients who received EPAG as front-line single treatment in our center, and 7 patients was enrolled from July, 2019 to September, 2020. Patients received EPAG 25mg to a maximum 75mg. The side effects, numbers of transfusion, improvement of peripheral blood and bone-marrow blast were recorded.

The 12weeks overall response rate was 85.7%, with 2 robust response (RR), and 4 hematologic response. In patients got response, 66.7% (4/6) became platelet transfusion independent, with an answering period from 19 days to 55 days. No drug adverse events were observed within 12 week-treatment, and no obvious increased blasts as well as myelofibrosis were observed. 2 patients repeated the next-generation sequencing, and no new mutations were found. After the 12-week treatment, the 2 RR patients were still taking EPAG and got continued complete remission with a median follow up of two years. Two patient (No.3 and No.4) relapsed after EPAG withdrawn and became transfusion depend again. Patient No.1 and No.6 died of infection and cerebral hemorrhage during the follow up.

In summary, EPAG may improve the hematopoiesis of TD-patient with low to intermediate risk-1 MDS, and was well tolerant with a dose of 25mg to 75mg for Asian population.

Figure 1 Figure 1.

Disclosures

No relevant conflicts of interest to declare.
T-Cell Phenotype Varies in Distinct Tumor Microenvironments and CD57 + T FH Cells Are Associated with Disease Progression and Inferior Survival in Follicular Lymphoma
Authors: Yang Zhi-Zhang , Kim Hyojin , Wu Hongyan , Tang Xinyi , Krull Jordan E. , Villasboas Jose , Novak Anne J. , Ansell Stephen M.
Published: November 2021

The tumor microenvironment plays a crucial role in mediating the tumor immune response, thereby affecting patient outcomes in follicular lymphoma (FL). Using CyTOF, we analyzed single cell suspensions created from pre-treatment biopsies in a cohort of 82 FL patients and retrospectively extracted patient clinical parameters. Hierarchical analysis of the tumor microenvironment composition stratified these patients into 4 groups: group 1 represents patients with high percentage of monocyte/macrophages/NK cells; patients from group 2 and 3 are rich of intratumoral T and B cells, respectively. Patients with intermediate number of T and B cells are classed into group 4. Intratumoral T cells from these 4 groups exhibited distinct phenotypical characteristics. Group 1 was enriched with KLRG1 +CD8 + T cells, while group 4 showed increased number of CXCR3 - T EM and CD4 + T N cells. Patients from group 2 and 3 featured with increased number of ICOS + T reg cells and CD57 + T FH cells, respectively. We observed that higher numbers of T cells significantly correlated with early disease stage, lack of B-symptoms, event-free survival at 24 months (EFS24) and a favorable overall and EFS survival in patients with histological grade 1 and 2. However, this association was not seen in patients with histological grade 3a and b, suggesting that T cell-mediated anti-tumor immunity is preserved in low grade patients, but overridden in higher grade patients. CITRUS analysis revealed that a cluster that is CD57 + and PD-1 high was significantly more abundant in patients who had disease progression than patients who had a complete response to therapy, suggesting a role of CD57 + T FH cells in promoting malignant cell growth in FL. Consistent with this finding, we observed that higher number of CD57 + T FH cells correlated with an inferior survival in FL. Using CITE-seq technology, we found that CD57 + T FH cells exhibited a substantially different transcriptome when compared to CD57 - T FH cells. Genes that were differentially upregulated in CD57 + T FH cells when compared to CD57 - T FH cells included genes involved in cell survival, compromised inflammatory response, and metabolism activation including GZMK, CCL4, CST7, DUSP2, LGALS3, CYTOR, CHI3L2, SYNE2, CXCL13, CD27 and FABP5. Taken together, our results indicate different tumor microenvironments among FL patient groups that is associated with variable T-cell phenotype. We found that CD57 + T FH cells play an important role in predicting disease progression and patient prognosis in FL.

Figure 1 Figure 1.

Disclosures

Novak: Celgene/BMS: Research Funding. Ansell: Bristol Myers Squibb, ADC Therapeutics, Seattle Genetics, Regeneron, Affimed, AI Therapeutics, Pfizer, Trillium and Takeda: Research Funding.
Subcutaneous Marzeptacog Alfa (Activated) Versus Intravenous rFVIIa for Treatment of Episodic Bleeding in FVIII Deficient Hemophilia a Rats
Authors: Knudsen Tom , Johansen Peter , Reckless Jill , Desai Shraddha , Blouse Grant E.
Published: November 2021

Background: FVIII deficient knock-out (F8 -/-) rats mimic the bleeding incidents seen in severe human hemophilia A (HA). Subcutaneous (SQ) marzeptacog alfa activated (MarzAA), a novel, engineered recombinant activated coagulation Factor VII (rFVIIa) has been shown to effectively treat episodic bleeding in a pilot experiment in HA rats. This study evaluated the effect of single SQ doses of MarzAA and a single intravenous (IV) dose of rFVIIa on episodic bleeding in F8 -/- rats. Moreover, it compared the effect of SQ MarzAA and IV rFVIIa directly.

Methods: Animals were allocated to treatment with either SQ vehicle, SQ MarzAA (60, 180 or 385 µg/kg) or IV rFVIIa (NovoSeven ®) (580 µg/kg) immediately after the bleeding was diagnosed. Doses were based on allometric scaling from humans (Nair AB and Jacob S. J Basic Clin Pharm 2016; 7: 27-31). The primary endpoint of the study was clinical efficacy as rated by a well-defined 4-point scale (Excellent, Good, Fair or Poor), and the efficacy assessment was either treatment success (Excellent or Good) or treatment failure (Fair or Poor) at the 24-hour timepoint. All personnel handling or assessing animals were blinded to the treatment status of each animal except those dosing animals who knew the route of administration.

Results: A total of 86 F8 -/- rats was enrolled in the study. Of these, 61 rats presented treatment eligible bleeds between 3 and 10 weeks of age. No statistically significant difference in bleeding severity score were found across groups on diagnosis. As assessed by the clinical outcome at 24 hours, all three SQ MarzAA dose groups exhibited a statistically significant effect on treatment response when compared to SQ vehicle treatment (Figure 1). Conversely, no statistically significant effect could be identified when the single IV rFVIIa 580 µg/kg dose group was compared to vehicle. The overall treatment success rates at 24 hours were: SQ vehicle: 8%, SQ MarzAA 60 µg/kg: 58%, SQ MarzAA 180 µg/kg: 67%, SQ MarzAA 385 µg/kg: 85%, and IV rFVIIa 580 µg/kg: 33%. When compared directly using allometric scaling of clinical doses, SQ MarzAA at 385 µg/kg exhibited a statistically superior effect compared to IV rFVIIa at 580 µg/kg (p=0.0154, Fischer's exact test).

Conclusion: Single doses of SQ MarzAA were effective in treating episodic bleeding in HA rats and statistically distinguishable from vehicle at all dose levels tested. When clinically relevant doses were compared directly to rFVIIa, SQ MarzAA compared favorably to IV rFVIIa. Taken together, these data provide robust nonclinical evidence that a single dose of SQ MarzAA may be successful in treating episodic bleeding when administered after bleeding has started.

Figure 1 Figure 1.

Disclosures

Knudsen: Catalyst Biosciences: Current Employment, Current holder of individual stocks in a privately-held company. Blouse: Catalyst Biosciences: Current Employment.
Genetic Screening of Patients with Evans Syndrome: A Single Centre Analysis
Authors: Miano Maurizio , Guardo Daniela , Grossi Alice , Palmisani Elena , Fioredda Francesca , Terranova Paola , Lupia Michela , Cappelli Enrico , Dell'Orso Gianluca , Lanciotti Marina , Ceccherini Isabella , Dufour Carlo
Published: November 2021

Background:

Evans syndrome (ES) is a rare disorder defined as the simultaneous or sequential presence of autoimmune haemolytic anemia and immune thrombocytopenia but it can also be considered as the presence of at least two autoimmune cytopenias. Recent reports have shown that ES is often a manifestation of an underlying congenital immune dysregulation syndrome that, in some case, can benefit from specific treatments.

Aims:

The aim of this study is to investigate the clinical/immnunological characteristics and the underlying genetic background of a single centre cohort of patients with ES.

Methods:

Data were obtained from a retrospective charts' review of patients with a diagnosis of ES in our centre. Genetic studies were performed with NGS analysis of 315 genes related to both hematologic and immunological disorders as congenital bone marrow failure syndromes, primary immunodeficiencies, and primary immune regulatory disorders.

Results:

Fourteen patients (23 males, 17 females) with a median age at onset of 6 years (range 0-16) were studied. ES was concomitant and sequential in 18 (45%) and 22 (55%) patients, respectively. Nine/40 (8%) patients had a family history of autoimmunity. Other abnormal immunological features and signs of lymphoproliferation were present in 24/40 (60%) and 29/40 (72%) of cases, respectively. Seventeen out of 40 (42%) children fitted the ALPS diagnostic criteria. The remaining 15 (37%) and 9 (22%) were classified as having an ALPS-like phenotype and an isolated ES, respectively. Twenty patients (50%) were found to have an underlying genetic defect on TNFRSF13B, FAS, CTLA4, IKBGK, CARD11, LIG4, LRBA, STAT3, CASP10 and ADA2 genes. Table 1 shows the details of clinical/immunological characteristics of patients with or without a genetic diagnosis. No significant differences were noted between the two groups.

Conclusions:

This study shows that half of patients with ES have a genetic background, secondary to Primary Immunodeficiencies. Therefore, an immunological screening and an extended molecular evaluation should be offered to all patients, since specific genetic diagnosis may benefit from targeted treatments.

Figure 1 Figure 1.

Disclosures

No relevant conflicts of interest to declare.
Online Patient-Reported Platform Detects Trend of Increased COVID-19 Risk and Severity for Multiple Myeloma Patients on Active Lenalidomide-Based Therapy
Authors: Liu Emily I. , Sweeney Nathan W. , Ahlstrom Jennifer M.
Published: November 2021

Background: Since the start of the COVID-19 pandemic, "high-risk" patients, such as cancer patients, have had to reconsider their current medical treatments and other treatment alternatives to best minimize their risk for contracting COVID-19. Lenalidomide has immunomodulatory properties that stimulate the production of T-cells which help combat against infections which may include diseases such as COVID-19. In this abstract, we investigate whether lenalidomide protects multiple myeloma (MM) patients from contracting COVID-19 and whether lenalidomide decreases the severity of COVID-19 events (including hospital or intensive care unit [ICU] admissions, and need of assisted ventilation) for patients that contract the virus (PMID: 32950467, PMID: 32353254).

Methods: MM patient data was collected through HealthTree ® Cure Hub for Multiple Myeloma (healthtree.org) and the relative risk was calculated to compare the risk of contracting COVID-19 between patients taking lenalidomide and those who were not at the time of contracting COVID-19. The odds ratio was calculated to measure lenalidomide's effect on the severity of COVID-19 if contracted. These events include whether a patient was hospitalized, had to go to the ICU, or required oxygen therapy.

Results: There were 1,123 patients involved in comparing lenalidomide with the risk of contracting COVID-19, including patients that never tested positive for COVID-19. Surprisingly, our results showed that patients who are taking lenalidomide have a 10% higher risk for contracting COVID-19 than those who are not; however, these findings were insignificant. Furthermore, 40 patients were involved in investigating lenalidomide's effect on decreasing severe COVID-19 symptoms for MM patients. Our results showed that the odds of patients experiencing severe COVID-19 were 1.95 times more for those on lenalidomide than those who were not.

Conclusions: Despite the insignificance of lenalidomide during COVID-19, our results indicated that taking lenalidomide may not be beneficial in lowering the risk for MM patients in contracting COVID-19. Furthermore, lenalidomide may also not decrease the severity of COVID-19 symptoms for MM patients that did contract COVID-19. Our results may help MM patients and their providers decide whether to continue their use of lenalidomide or to seek alternative treatment options.

Disclosures

Ahlstrom: Pfizer: Other: Patient Advisory; Janssen: Other: Patient Advisory; Takeda: Other: Patient Advisory; Bristol Myers Squibb: Other: Patient Advisory.
Activity of Limited-Schedule Bendamustine and Methylprednisolone in Chronic Lymphocytic Leukemia
Authors: Quan Walter JR , Kadakia Meet , Krishnan Nithya , Stack Anthony , Rogers Christy , Quan Francine Marie
Published: November 2021

Bendamustine was approved by the United States Food and Drug Administration in 2008 for the treatment of Chronic Lymphocytic Leukemia (CLL). Corticosteroids have been administered in regimens with chemotherapy for CLL for more than 40 years. Despite this, the combination of Bendamustine and Methylprednisolone has not been explored in CLL. We describe our experience with Bendamustine 100 mg/m2 and Methylprednisolone 125 mg IV on days 1 and 2 and Peg-filgastrim 6 mg subcutaneously on day 3 every 28 days for 3-4 cycles in 12 patients with CLL. Cycles have been limited to attempt to prevent cumulative marrow toxicity due to Bendamustine. Patient characteristics: 7 males/5 females, median age-58, Binet stage B (8), C (2), A (2); ZAP70/CD38+ (5), Trisomy 12 (3), 13q- (2), normal cytogenetics (2); median ECOG status 1, no prior therapy in 8. Median number of cycles received = 3 (two received 4 cycles). Most common toxicities: nausea/emesis (5), fever/flu-like symptoms (4), arthralgia/myalgia (4). There have been no treatment-related deaths. One patient was briefly hospitalized for pneumonia. Complete responses have been seen in 6 patients (median duration = 50.7 months; range: 12.1+ to 87.3 months). Partial responses have been seen in 4 patients (median duration = 17.3+ months; range: 3.3 to 26.9+ months). The combination of Bendamustine and Methylprednisolone given in a limited schedule is well-tolerated and is active in CLL.

Disclosures

No relevant conflicts of interest to declare.
Post-Transfusion Takotsubo Cardiomyopathy
Authors: Badami Krishna G
Published: November 2021

In three recent cases an initial transfusion-related adverse event (TRAE) occurred that had a significant respiratory component, and other features suggesting a severe allergic reaction. All were treated as such. Shortly after treatment the nature of the event changed to something resembling transfusion-associated circulatory overload (TACO).

These cases suggest the possibility that a severe TRAE that is not TACO may precipitate TACO. In other words, some transfusions, sometimes, may deliver a 'double whammy'. A possible mechanism might be through the development of Takotsubo, or stress, cardiomyopathy - a cause of acute, reversible, heart failure. Takotsubo cardiomyopathy typically presents with chest pain or shortness of breath. Ventricular wall dysfunction occurs, and characteristic echocardiographic abnormalities are seen. ECG and biochemical features suggest cardiac ischaemia. It is commoner in post-menopausal women. A range of 'stressors' - hypotension, asthma attacks, accidents, medical procedures, pain, bad news, etc. - may precipitate it. 1 Catecholamines such as adrenaline are believed to play a central role. They cause direct toxicity to the myocardium, and also coronary artery spasm, and increased cardiac workload. 2 One other such case is reported. There, a 48 years old post-menopausal woman developed acute reversible heart failure with the features of Takotsubo cardiomyopathy, following urticaria and pruritus, whilst having a platelet transfusion. No adrenaline was given, but the authors postulate that adrenergic and histaminergic elements may have combined to produce the effect on the heart. 3 Thus, in some patients the 'perfect storm' compounded of borderline cardiac function,catecholamine release (secondary to the stress of the underlying disorder, the transfusion, and the TRAE),histamine release (in allergic TRAE),the treatment of the allergic reaction with adrenalinerecent or ongoing volume overload

may be sufficient to precipitate or exacerbate TACO. Careful consideration of cases with similar features is needed. Potential precipitating factors are modifiable or preventable.

References:

1. https://www.health.harvard.edu/heart-health/takotsubo-cardiomyopathy-broken-heart-syndrome

2. https://www.ahajournals.org/doi/full/10.1161/CIRCULATIONAHA.116.027121

3. Zhou JQ, Choe E, Ang L, et al. Stress-induced cardiomyopathy associated with a transfusion reaction: A case of potential crosstalk between the histaminic and adrenergic systems. Exp Clin Cardiol.2011; 16: 30-32

Figure 1 Figure 1.

Disclosures

No relevant conflicts of interest to declare.
PDGF-BB and Its Role in Megakaryopoiesis and Thrombocythemia
Authors: Yang Mo , Li Liang , Luo Yi , Su Weiqing , Kong Huimin , Chong Beng H , Ye Jieyu
Published: November 2021

Background: Essential Thrombocythemia (ET) is characterized by persistently elevated platelet counts in the context of a normal red cell mass. However, the molecular mechanism of ET is still under investigation. Our previous studies demonstrated the presence of functional PDGF receptors (PDGFR) on megakaryocytes and their ability to mediate hematopoiesis and megakaryopoiesis (Ye et al, Haematologica, 2010). The role of PDGF-BB on megakaryocytic progenitors (CD41+, CD34+ cells) and its mechanisms on ET will be further studied in this project.

Methods: ELISA, CFU assay, immunofluorescence microscope, flow cytometry and NOD/SCID mice were used in this study.

Results: Bone marrow plasma levels of PDGF-BB in ET patients (n=18) and normal control (n=10) were tested and found an increased PDGF-BB levels in ET patients (2071.2±124.8 pg/ml), compared with normal control (1382.5±128.3pg/ml) (P=0.002). In vitro experiment, PDGF-BB promoted the ex vivo expansion of human hematopoietic stem (CD34 +) and progenitor (CD41 + CD61 +) cells. More significantly, PDGF enhanced the engraftment of human CD45 + cells and their myeloid subsets (CD33 +, CD14 + cells) in NOD/SCID mice. PDGF-BB stimulated megakaryopoiesis via PDGFR and its signalling. It also showed a direct stimulatory effect of PDGF-BB on c-Fos, GATA-1 and NF-E2 expressions in megakaryocytes. We speculate that these transcription factors might be involved in the signal transduction of PDGF-BB on the regulation of megakaryopoiesis. PDGF-BB also enhanced platelet recovery in mice model with radiation-induced thrombocytopenia. Studies showed that PDGF, like TPO, significantly promoted platelet recovery and the formation of CFU-MK in this irradiated-mouse. An increased number of hematopoietic stem/progenitor cells and a reduction of apoptosis were found in the bone marrow histology sections. In the CHRF apoptotic model, PDGF-BB had a similar anti-apoptotic effect as TPO on megakaryocytes. We also demonstrated that PDGF-BB activated the p -Akt , p-Jak2 and p-Stat3 expression, while addition of imatinib mesylate reduced p-Akt, p-Jak2 and p-Stat3 expression in CHRF cells.

Conclusion: Our findings suggested that PDGF-BB is likely to be mediated via PDGF receptors with subsequent activation of the Akt and Jak2/ Stat3 pathways in megakaryopoiesis. These studies provide a possible explanation that PDGF-BB and its signaling may be involved in the molecular mechanism of essential thrombocythemia.

Disclosures

No relevant conflicts of interest to declare.
Genomic Analysis of Cutaneous CD30-Positive Lymphoproliferative Disorders
Authors: Abdulla Farah R , Zhang Weiwei , Wu Xiwei , Honda Kord , Qin Hanjun , Cho Hyejin , Querfeld Christiane , Zain Jasmine , Rosen Steven T. , Chan Wing C. , Parekh Vishwas , Song Joo Y.
Published: November 2021

Primary cutaneous CD30+ T-cell lymphoproliferative disorders (CD30+ LPD) are the second most common cutaneous lymphomas. According to the World Health Organization (WHO), CD30+ LPD include primary cutaneous anaplastic large cell lymphoma (pcALCL) and lymphomatoid papulosis (LyP) as well as borderline lesions. pcALCL and LyP is thought to represent two ends of a spectrum of diseases that have different clinical presentations, clinical courses, and prognoses in their classic forms, but share the same histology of medium to large CD30+ atypical lymphoid cell infiltrates. Because the behavior of these entities is different clinically and prognostically, we aim to search for oncogenic genomic variants using whole exome sequencing (WES) that drive the development of LyP and pcALCL.

Clinical information, pathology, immunohistochemistry, and T-cell rearrangements on six cases of LyP and five cases of pcALCL were reviewed to confirm the rendered diagnosis prior to WES of all specimens. All cases of CD30 LPD had recurrent mutations in at least one of the epigenetic modifying genes, with the most frequent mutations found in the mixed lineage methyltransferase family involved in the methylation of H3K4: CREBBP (27%), KMT2A (36%), KMT2D (36%), SETD2 (27%), and SMARCA4 (27%) (Figure). Within the JAK/STAT pathway, mutations of STAT3 were observed in 2 cases of pcALCL (n=2, 18%), STAT5B in one case of LyP (n=1, 9%) and JAK1 mutation in one case of LyP(n=1, 9%). Lastly, mutations were also identified in the T-cell signaling pathway. 3/5 cases of pcALCL demonstrated loss of function mutations in EOMES, a T-box transcription factor important in lymphocyte development. We had two cases of C-ALCL and one case of LyP with NOTCH1 mutations supporting the importance of this gene in T-cell lymphomas.

TP53 mutations and copy number loss is more characteristic of aggressive lymphomas and these abnormalities were absent in pcALCL and LyP and may explain the indolent nature of CD30+ LPD.

While the JAK/STAT pathway, T-signaling, and epigenetic alterations possibly play a role in the pathogenesis of these diseases, genes involved in cell-cycle control and apoptosis (ie. TP53) that are characteristic of more aggressive diseases were not identified. Extending this investigation to a larger number of samples will allow us to detect additional mutations that may help distinguish between CD30+ LPDs of LyP and pcALCL as well as other histologic mimics such as systemic ALCL and large cell transformation of MF.

Figure 1 Figure 1.

Disclosures

Abdulla: Caris Life Licenses: Current Employment. Zain: Kiyoaw Kirin, Secura Bio, Seattle Genetics: Honoraria; Secura Bio, DaichiSankyo, Abbvie: Research Funding; Secura Bio, Ono , Legend, Kiyowa Kirin, Myeloid Therapeutics Verastem Daichi Sankyo: Consultancy.
Characterising Tumour and Microenvironmental Responses to R-CHOP in Immunocompetent Mouse Models of DLBCL
Authors: Maybury Bernard D , Bewicke-Copley Findlay , Johnson Peter , Fitzgibbon Jude , Calado Dinis Pedro
Published: November 2021

R-CHOP chemoimmunotherapy has been the standard of care for diffuse large B cell lymphoma (DLBCL) for 20 years. The lymphoma and microenvironment responses to R-CHOP have not been studied in detail in an experimental setting. Here we describe the early effects of R-CHOP on lymphoma cells and infiltrating T cells in novel mouse models of DLBCL. Using cre-lox recombination to induce B cell-specific overexpression of MYC, IKK2 and BLIMP1 drives the formation of unique high grade B cell lymphomas with an activated B cell (ABC) phenotype and an intact immune environment. Using flow cytometry and RNA sequencing we compare the response to R-CHOP in these mice with the response in a cell line lymphoma model. We also performed partial splenectomy prior to R-CHOP treatment to study paired samples pre-treatment and at relapse by whole-exome and RNA sequencing. Treatment with anti-CD20 alone induced very few changes in lymphoma cells in either model, but CHOP chemotherapy in combination with rituximab induced changes in lymphoma cell and T cell phenotypes. In particular, extracellular matrix and cell adhesion gene signatures were enriched in lymphoma cells after treatment, in both disease models studied. After injection of the cell line, memory populations of CD8+ T cells are expanded but this change is reversed by chemotherapy, whereas in the cre-lox conditional model 40% of CD8+ T cells are exhausted with smaller chemotherapy-associated changes. In the paired pre-treatment/relapse samples we observed clonal selection and increased mutation burden at relapse, associated with diverse transcriptional changes. Mutations were newly detected or expanded at relapse in various lymphoma-associated genes including Trp53, Samhd1 and Ubr5. Our results suggest that DLBCL responses to treatment and biology at relapse are principally driven by tumour-specific factors, but there are some commonalities across model systems which may be amenable to therapeutic modulation. The differences in T cell responses between conditional tumours and cell-line tumours demonstrate the limitations of cell lines for studying treatment responses in vivo.

Figure 1 Figure 1.

Disclosures

Johnson: Boehringer Ingelheim: Consultancy; Janssen: Consultancy; Kite Pharma: Honoraria; Oncimmune: Consultancy; Epizyme: Consultancy, Research Funding; Bristol-Myers: Honoraria; Incyte: Honoraria; Genmab: Honoraria; Takeda: Honoraria; Novartis: Honoraria; Morphosys: Honoraria; Celgene: Honoraria; Kymera: Honoraria. Fitzgibbon: Epizyme: Research Funding. Calado: Myricx Pharma: Consultancy, Current holder of individual stocks in a privately-held company, Current holder of stock options in a privately-held company, Patents & Royalties: Cancer Treatments. WO patent WO 2020/128475 A1 (2020).
A Snapshot of Venous Thromboembolism (VTE): Retrospective Audit of Events at a Metropolitan Tertiary Referral Hospital
Authors: Chong Julianne , Dissanayake Rukmal K , Younan Sarah , Chan Michael V , Yang Yao , Wong Doo Nicole , Chen Vivien
Published: November 2021

Background

Facility-level review of hospital-acquired venous thromboembolism (HA-VTE) cases, including pulmonary embolism (PE) and deep vein thrombosis (DVT), can provide insight into the local drivers of this preventable complication.

Aim

To determine the nature of HA-VTE, patient characteristics, VTE risk assessment completion and appropriateness of prescribed VTE prophylaxis at a metropolitan tertiary referral hospital and to compare the number of HA-VTE detected by audit of imaging scans with those reported by clinical coding.

Methods

We conducted a retrospective electronic medical record audit encompassing all patients diagnosed with HA-VTE via ventilation-perfusion scan, computed tomographic pulmonary angiography and/or doppler ultrasound using a demographic and VTE risk assessment audit tool during selected months of 2017-2019. We compared the number of manually audited cases with the number of performance unit-coded cases for the same months.

Results

The months of May 2017, October 2017, April to July 2018 and April to July 2019 were included. There was a significant difference between HA-VTE detected via manual audit (147 events) and hospital coding (18 events), p=0.002. Manual audit patients were majority non-surgical (65%), female (58%), over 60 years (80%), at moderate VTE risk (71%) with reduced mobility (52%). There were 108 DVT-only (73%), 23 PE-only (16%) and 16 DVT plus PE events (11%). Notable risk factors were moderate to major surgery (30%), active malignancy or cancer treatment (24%) and active infection (27%). Most patients were prescribed appropriate VTE prophylaxis (74%) and had documented VTE risk assessment (68%). Appropriate VTE prophylaxis was significantly associated with using an electronic clinical decision support tool during VTE risk assessment (p=0.024).

Conclusion

Facility-driven HA-VTE audits provide opportunity to developed targeted initiatives for at-risk patients at a local level. Greater numbers of HA-VTE identified via manual audit suggest the need for future real-time documentation of HA-VTE to assist with efficient case review.

Disclosures

No relevant conflicts of interest to declare.
Neutrophil Extracellular Traps Promote Joint Injury in Hemophilia
Authors: Kaminski Tomasz W. , Brzoska Tomasz , Tutuncuoglu Egemen , Ragni Margaret V. , Sundd Prithu
Published: November 2021

Epidemiological evidence suggests that recurring episodes of joint-bleeding contribute to the development of hemophilic arthropathy (H) in 70-85% of hemophilia patients. Despite major advances in the treatment to prevent joint bleeding, HA continues to be a major morbidity affecting hemophilia patients and the etiological mechanism contributing to the progression of HA remains poorly understood. Recent evidence suggests that the accumulation of blood in the joints may lead to the release of erythrocyte-derived DAMPs (eDAMPs) such as heme and hemoglobin that can promote sterile inflammation, however, the innate immune pathways contributing to this pathophysiology remain unknown.

In the study, we used a model of puncture-induced knee joint injury in FVIII-total knockout (F8TKO) mice and blood samples from hemophilia-A patients diagnosed with HA. Intravital multi-photon-excitation fluorescence intravital (in vivo) microscopy of injured synovium in live F8TKO or control mice was conducted to assess neutrophil-platelet aggregation and NETs generation in the knee-joint. Imaging-flow-cytometry and ELISA assays were used to estimate the number of circulating NETs in plasma of patients diagnosed with HA and mice after the knee-injury procedure. Scoring of the bleeding severity, histology, IHC and confocal imaging of joints were conducted to quantify the joint injury in mice.

F8TKO but not control mice manifested knee-joint injury and severity of bleeding 5-days post knee-injury. Progression of knee-joint injury was associated with increased neutrophil accumulation and NETs shedding within the synovium of F8TKO mice. Circulating NETs were significantly abundant in the plasma of hemophilia patients diagnosed with HA and F8TKO mice following knee-injury but not plasma of control humans or mice.

These findings are the first to suggest that NETs contribute to pathogenesis of HA in hemophilia. Currently, experiments are underway to identify the innate immune pathways that promote NETs shedding, leading to joint-damage in hemophilia.

Disclosures

Ragni: Takeda Therapeutics: Membership on an entity's Board of Directors or advisory committees; Bioverativ (Sanofi): Membership on an entity's Board of Directors or advisory committees; BioMarin Pharmaceutical: Membership on an entity's Board of Directors or advisory committees; Alnylam (Sanofi): Membership on an entity's Board of Directors or advisory committees; University of Pittsburgh: Research Funding; Spark Therapeutics: Membership on an entity's Board of Directors or advisory committees. Sundd: CSL Behring Inc: Research Funding; Novartis: Membership on an entity's Board of Directors or advisory committees, Research Funding; Bayer: Research Funding.
Screening Chest CT Prior to Allogenic Transplantation - High Rates of Occult Abnormalities
Authors: Alhomoud Mohammad , Gomez-Arteaga Alexandra , Mayer Sebastian A. , Hsu Jingmei , Phillips Adrienne A. , Shore Tsiporah B. , van Besien Koen
Published: November 2021

Introduction: Pulmonary complications constitute a major cause of morbidity and mortality in the post-hematopoietic cell transplant (HCT) period. While Chest X-ray (CXR) is customarily used for screening, we have utilized chest computed tomography (CT) within one month of transplant. Here we aim to characterize the prevalence of abnormalities and their impact on the eligibility for allogenic (allo) HCT and outcomes post-transplant.

Methodology: We conducted a single center retrospective study of all patients who were evaluated for allogenic HCT from January 2013 through December 2020 in New York Presbyterian Hospital - Weill Cornell Medicine (NYP-WCM). All patients who had chest CT as part of their pre-transplant evaluation were included for analysis.

Results: We identified 478 patients who had Chest CT screening. In 396 CT chest was normal or confirmed previous abnormalities. Eighty-two patients (17%) had previously undetected abnormalities (Figure 1). The most frequent abnormalities were pulmonary nodules (defined as nodules of 4 mm or greater) in 27 patients (31%), ground-glass opacities (GGO) in 21 patients (25%), Pneumonia in 18 patients (21%). Miscellaneous findings not related to the primary disease were found in 12 patients (14%) including thyroid nodules, breast nodules and hemangioma of the liver. A new malignancy was found in 6 patients (7%) and incidental pulmonary embolism (PE) in 2 patients (2%) (Figure 2). There were 5 (6%) patients who were ultimately excluded from transplant due to CT chest findings (simultaneous CXR showed abnormalities in only 1 out of 5). Three of those patients were found to have invasive fungal infection, and the other two had unrecognized metastatic lung cancer. For 19 patients (23%), transplant was delayed for diagnostic procedure and/or treatment of pulmonary findings (CXR showed abnormalities in only 3 patients out of 19). The most common reason for delay was lung infection requiring treatment. Thirty-two patients (41%) out of 77 patients with abnormal CT scan who eventually underwent transplant, have died . Sixteen died after relapse, and 16 from non-relapse mortality (NRM) with pulmonary complications playing a role in 13 patients (Figure 3).

Conclusion: 17% of patients who had a pre-transplant CT chest had abnormalities that warranted further evaluation. In 23% of these patients, these findings led to a delay in transplant for further optimization. Six percent were deemed ineligible for transplant due to absolute contraindications that were incidentally discovered on chest CT. Initial screening CXR failed to identify a significant number of abnormalities. Our data suggests that chest CT imaging should be part of the routine pre-transplant evaluation.

Figure 1 Figure 1.

Disclosures

No relevant conflicts of interest to declare.
Neutrophil Metabolic Rewiring in Severe and Mild COVID-19
Authors: Li Yafeng , Hook Jessica S , Ding Qing , Chung Stephen , Moreland Jessica , Agathocleous Michalis
Published: November 2021

SARS-CoV-2 infection can cause severe respiratory disease resulting in hospitalization and death in some people, but in others it causes a mild disease or is asymptomatic. The causes of severe COVID-19 are of profound public health importance. Severe COVID-19 is marked by an exacerbated innate immune response leading to hyperinflammation which damages host tissues. Broad inhibition of hyperinflammation by dexamethasone is a mainstay of severe COVID-19 treatment. However, more targeted therapeutic approaches to dampen hyperinflammation are lacking. Patients typically exhibit increased neutrophil numbers in the blood and lung, and increased formation of neutrophil extracellular traps. Previous work profiled the metabolome of plasma in severe COVID-19 patients and found several metabolic aberrations as compared to mild COVID-19 patients or healthy controls. However, the metabolome of immune cell populations in COVID-19 has not been characterized to our knowledge. In addition, it is not known if metabolic changes observed in severe COVID-19 mechanistically contribute to hyperinflammation and host tissue damage or if they are simply markers of pathology.

To understand how metabolism can regulate the innate immune system in COVID-19 and to explore possible therapeutic targets, we carried out a comprehensive metabolomic study of neutrophils purified from healthy controls, mild/moderate COVID-19 patients, and severe COVID-19 patients. Severe COVID-19 patients were defined as having Acute Respiratory Distress and being treated in the intensive care unit. Plasma from the same patients and healthy controls was also collected and metabolomics was performed to study how COVID-19 impacts the neutrophil and plasma metabolome in tandem.

Several metabolic pathways were significantly different between mild COVID-19 patients, severe COVID-19 patients, and healthy controls, consistent with rewired neutrophil and plasma metabolome in severe COVID-19. Differentially abundant metabolites belonged to central carbon metabolism, nucleotide metabolism, amino acid metabolism and other pathways. Variation in the PMN metabolome was largely independent from variation in the plasma metabolome. This suggests that processes internal to neutrophils drive changes in neutrophil metabolism. Metabolomics in 3 mouse models of hyperinflammation showed common or distinct metabolic features that change in inflammation compared to metabolomics of severe COVID-19 patients. Therefore changes in some metabolites comprise a general hyperinflammatory metabolic signature while others are more specific to severe COVID-19. Isotope tracing experiments were carried out to understand how nutrients were utilized in neutrophils in context of inflammation. We will describe mechanistic work testing whether metabolic features we observe in severe COVID-19 patients regulate the inflammatory response of human neutrophils.

Disclosures

No relevant conflicts of interest to declare.
Central Line Associated Bloodstream Infection Rate Among Hematologic Malignant Patients Who Received Antimicrobial Prophylaxis in the Inpatient Setting
Authors: Abbagoni Sujith , Le Thuy , Bryan Locke J. , Mian Mohammad A. H. , Bradshaw Danielle , Fox Ashley D , Jillella Anand P , Okereke Chijioke , Kolhe Ravindra , Savage Natasha M. , Kota Vamsi
Published: November 2021

Introduction:

Patients with hematological malignancies are at increased risk of infections which leads to increased morbidity and mortality. Particularly, they are at an increased risk of developing central line associated bloodstream infection (CLABSI) since they frequently require central venous catheters for various essential reasons including chemotherapy, blood transfusions, and parenteral nutrition. Prophylaxis with antibiotics is the key in the management for these patients but is not always effective and sometimes associated with complications. Hence, the main objective of the study is to examine the rate of developing CLABSI in patients with hematological malignancies receiving antimicrobial prophylaxis and in those who did not receive prophylaxis.

Methods

We conducted a retrospective study involving patients diagnosed with a hematological malignancy admitted to the Augusta University Medical Center from 2002 to 2019. IRB approval was obtained to review the patients' charts. Patients who developed CLABSI are defined as those who had positive blood cultures after central venous catheter had been placed. Percentage of patients who received prophylaxis and those who didn't get antimicrobial prophylaxis were calculated. The difference between the two percentages was examined to determine whether or not it is statistically significant.

Results

The charts of 268 patients admitted for hematological malignancies were reviewed. Among 173 patients receiving antibiotic prophylaxis (65.0%), 78 (45.1%) developed CLABSI. Among the patients who didn't receive prophylaxis, 64 (68.8%) developed CLABSI. The difference between the two groups was statistically significant (p<0.01), and the relative risk of developing CLABSI in patients who did not receive prophylaxis is 0.57 (95% confidence interval 0.41 - 0.79).

Conclusion

Patients who had been diagnosed with a hematologic malignancy and admitted at the academic institution from 2002 to 2019 were analyzed for the risk of developing CLABSI that was associated with the use of antibiotic prophylaxis. We found that the non-prophylactic group has statistically significant higher risk of developing CLABSI than the prophylactic group. Thus prophylaxis with antibiotics may play a major role in decreasing the rate of CLABSI in patients admitted with hematological malignancy.

Disclosures

No relevant conflicts of interest to declare.
Systematic analysis of CCCH zinc finger family in Brassica napus showed that BnRR-TZFs are involved in stress resistance
Authors: Pi Boyi , Pan Jiao , Xiao Mu , Hu Xinchang , Zhang Lei , Chen Min , Liu Boyu , Ruan Ying , Huang Yong
Published: November 2021

Background

CCCH zinc finger family is one of the largest transcription factor families related to multiple biotic and abiotic stresses.Brassica napusL., an allotetraploid oilseed crop formed by natural hybridization between two diploid progenitors,Brassica rapaandBrassica oleracea. A systematic identification of rapeseed CCCH family genes is missing and their functional characterization is still in infancy.

Results

In this study, 155CCCHgenes, 81 from its parentB. rapaand 74 fromB. oleracea, were identified and divided into 15 subfamilies inB. napus. Organization and syntenic analysis explained the distribution and collinearity relationship ofCCCHgenes, the selection pressure and evolution of duplication gene pairs inB. napusgenome. 44 diploid duplication gene pairs and 4 triple duplication gene groups were found inB. napusof CCCH family and the segmental duplication is attributed to mostCCCHgene duplication events inB. napus. Nine types of CCCH motifs exist inB. napusCCCH family members, and motif C-X7/8-C-X5-C-X3-H is the most common and a new conserved CCH motif (C-X5-C-X3-H) has been identified. In addition, abundant stress-related cis-elements exist in promoters of 27 subfamily IX (RR-TZF) genes and their expression profiles indicated thatRR-TZFgenes could be involved in responses to hormone and abiotic stress.

Conclusions

The results provided a foundation to understand the basic characterization and genes evolution ofCCCHgene family inB. napus, and provided potential targets for genetic engineering in Brassicaceae crops in pursuit of stress-tolerant traits.