bluuebunny/crossref_metadata_2025 huggingface dataset:
Gls-010, a Novel Anti-PD-1 Mab in Chinese Patients with Relapsed or Refractory Classical Hodgkin Lymphoma: Preliminary Impressive Results of a Phase II Clinical Trial
Authors: Song Yuqin , Zhu Jun , Lin Ningjing , Zhang Mingzhi , Bai Hai , Liu Hui , Cui Jie , Ke Xiaoyan , Zhang Huilai , Liu Lihong , Yan Dongmei , Jiang Yongsheng , Zang Aimin , Qi Junyuan , Wang Li , Liu Zhuogang , Xu Bing , Zhang Ying , Zhang Zhihui , Meng Haijin
Published: November 2020

Introduction: Classical Hodgkin lymphoma (cHL) is characterized by genetic alterations in 9p24.1, leading to overexpression of PD-L1 ligand. GLS-010 is a novel fully human anti-PD-1 monoclonal antibody (mAb) and exhibited favorable results in previous Phase I study. The aim of this study was to evaluate the safety and efficacy profile of GLS-010 in Chinese patients (pts) with relapsed or refractory cHL.

Methods: In this multi-center, single-arm Phase II clinical trial, pts with relapsed or refractory cHL after at least 2 lines of prior systemic chemotherapies were enrolled and treated with GLS-010 240mg every 2 weeks until disease progression, death, unacceptable toxicity or withdraw from the study. Efficacy was assessed with the primary endpoint of objective response rate (ORR) by independent review committee (IRC) per Lugano 2014. Adverse events (AEs) were graded by NCI CTCAE v4.03.

Results: As of April 18, 2020, a total of 85 pts with relapsed or refractory cHL received a median of 14.5 treatment cycles (1 cycle include 2 injections). The pretreatment characteristics of the pts are shown in Table 1. At a median follow-up of 15.8 months, 28.2% (24/85) of pts discontinued treatment. As shown in Table 2, treatment-related adverse events (TRAEs) of any grade occurred in 79 (92.9%) of 85 patients, most of which were Grade 1-2. The most common TRAEs were fever (27/85, 31.8%), neutrophil count decreased (17/85, 20%) and alanine aminotransferase (ALT) increased (17/85, 20%). Grade ≥3 TRAEs occurred in 25 (29.4%) pts, most commonly, hepatic function abnormal (5/85, 5.88%) and hyperuricaemia (4/85, 4.71%). For all the 85 pts, ORR was 90.59% (77/85, 95%CI: 82.30-95.85) with 32.9% (28/85) of patients achieving a CR and 57.6% (49/85) of patients achieving a PR (Table 3). Median duration of response (DoR) and progression free survival (PFS) were not reached yet.

Conclusions: GLS-010 showed impressive anti-tumor activity (ORR=90.59%) and manageable safety profile in Chinese patients with relapsed or refractory cHL, and could be a new safe and effective treatment option.

Disclosures

Meng: Guangzhou Gloria Biosciences Co., Ltd.: Current Employment.
In-Silico Transcriptome Analyses of Hemostasis Triggers in Inflamed Vs Normal Mucosa of IBD Patients
Authors: Branca Enrica , Carrà Giovanna , Russo Isabella , Terzolo Massimo , Guerrasio Angelo , Morotti Alessandro
Published: November 2020

Background:inflammatory bowel disease (IBD) is characterized by chronic inflammation associated with an increased tendency to thrombosis and thromboembolic complications. The underlying mechanisms of VTE are not yet fully understood. Several studies reported different expressions of circulating procoagulant factors or fibrinolysis inhibitors in IBD. Others linked podoplanin overexpression with thrombosis, hypercoagulability, and increased risk of VTE. Here, we aimed to identify, among genes able to trigger thrombosis, those aberrantly expressed in IBD samples as compared with matched normal mucosa.

Methods:we analyzed the transcriptome of matched normal and inflamed lesions in 168 patients with Adult Crohn Disease (CD) and 245 pediatric IBD using publicly available datasets. We intentionally assessed the expression levels of triggers and inhibitors of the coagulation system (Tissue Factor, TFI), the fibrinolytic system (PLAU, PLAT, PAI-1/2/3, TAFI, PN-1, SERPINF2, A2M, SNX1, SERPINC1) and platelets activation (podoplanin). Analyses were finally performed using additional datasets with 219 ulcerative colitis (UC) adult patients, 198 IBD patients and compared to normal colon transciptome.

Results:in all datasets, when compared to matched normal mucosal transcriptome, CD and UC inflamed tissues over-expressed Podoplanin mRNA (p=2.46e-25). TF mRNA was also consistently up-regulated in inflamed mucosal of IBD patients, even if paralleled with up-regulation of TFPI as well, questioning on the functional role of TF in IBD. Among the regulators of the fibrinolytic system the urokinase-activator of the plasminogen is consistently deregulated in inflamed areas.

Conclusion: our in-silico analyses on gene expression profiles, using matched normal and pathological mucosa of the same patients, suggested that IBD inflamed mucosa favors platelets activation due to podoplanin over-expression. These observations require further studies to assess the biological role of podoplanin in IBD patients and opens new insights on how to perform antithrombotic prophylaxis in IBD patients.

Disclosures

No relevant conflicts of interest to declare.
Impact of Human T-Cell Lymphotropic Virus Infection on Survival Following Stem Cell Transplantation
Authors: Gupta Vishal K , El-Jawahri Areej , Janakiram Murali , Johnson Mariam H , Weinberg Janice , Goodrich Courtney B , Okoev Grigori , Sloan John Mark
Published: November 2020

Human T-Cell Lymphotropic Virus (HTLV) Types 1 or 2 are delta retroviruses that may lead to complications including adult T-cell leukemia-lymphoma (ATLL) and HTLV-1-associated myelopathy (HAM). There is concern that immunosuppression may result in progression from an asymptomatic carrier state to ATLL or HAM. Data on the safety of stem cell transplantation (SCT) in patients with HTLV-1/2 infection are lacking. The Center for International Blood and Marrow Transplant Research (CIBMTR) was queried for all patients who tested positive for HTLV infection in the pre-transplantation workup and underwent either autologous SCT (AutoSCT) or allogeneic SCT (AlloSCT) between 2008-2017. The primary outcome was overall survival up to four years post-SCT, assessed using the Kaplan-Meier statistic. All analyses were univariate and did not adjust for differences in disease, disease stage or patients characteristics. In those who underwent AutoSCT, 54 (0.5%) patients were HTLV-positive and 9,837 were HTLV-negative. In those who underwent AlloSCT, 126 (0.7%) patients were HTLV-positive and 18,084 were HTLV-negative. In the HTLV-negative group, a larger proportion underwent AlloSCT for acute myeloid leukemia in the HTLV-negative group, while Non-Hodgkin lymphoma/other leukemia was more common in the HTLV-positive group (P<0.001). Additional patient demographics are listed in Table 1. No difference in overall survival (p=0.661) or non-relapse mortality (p=0.522) was noted following AutoSCT based on HTLV status. Following AlloSCT, univariate analysis showed the HTLV-positive patients had a statistically significant lower overall survival (Figure 1; p=0.013) and higher non-relapse mortality at year four (Figure 2; p=0.023). In conclusion, survival after AutoSCT seems unaffected by HTLV status. While the underlying indications for AlloSCT differed in HTLV-infected patients, decreased overall survival and increased non-relapse mortality were seen in HTLV-positive patients. Reasons for increased non-relapse mortality may include increased infectious complications or graft-versus-host disease. Further studies are needed to understand the relative contributions of these factors.

Disclosures

Janakiram: Takeda, Fate, Nektar: Research Funding. Sloan:Stemline: Consultancy; Abbvie: Consultancy.
An Immune Risk Score Predicts Survival of Patients with Myelodysplastic Syndrome
Authors: Liang Yang , Hu Fang , Dai Yu-Jun , Wang Yun , Li Huan
Published: November 2020

Background: Myelodysplastic syndrome (MDS) was characterized as ineffective hematopoiesis, increased transformation to acute myeloid leukemia (AML), and accompanied by immune system dysfunction. However, the immune signature of MDS remains elusive.

Methods: The clinical data (age, sex, international prognostic score system (IPSS), hemoglobin, blast, RBC transfusion dependence, and corresponding subject-level survival) as well as expression profiles of MDS (CD34+ cells) were obtained from Gene Expression Omnibus (GEO: GSE 58831; GSE 114922). A robust prognosis model of immune genes was constructed by the least absolute shrinkage and selection operator (LASSO) regression analysis. Survival analysis for prognostic model was carried out through the Kaplan-Meier curve and Log-rank test. The receiver operating characteristic (ROC) curves and area under the curve (AUC) were used to assess the accuracy of prognostic models. Immune score for different subtype were calculated further by single sample gene set enrichment analysis (ssGSEA).

Result: A novel robust immune gene prognostic model indicate that subtype with lower risk score were longer overall survival (OS) than subtype with higher risk score in training cohort (Figure1 A, C). The model was further verified by the validation cohort (Figure1 B, D). The multivariate Cox regression analysis demonstrated the model was an independent prognostic factor for OS prediction with hazard ratios of 56.694 (95% CIs: 9.038−355.648), 3.009 (95% CIs: 1.042−8.692) both in train cohort and external validation cohort respectively (Figure1 G, H). The AUC of 5- year were 0.92 (95% CIs: 0.86 - 0.97) and 0.7 (95% CIs: 0.51 - 0.89) for OS respectively in training cohort and validation cohort (Figure1 E, F). Furthermore, ssGSEA showed higher risk score subtype was significantly associated with higher immune score of check point, human leukocyte antigen (HLA), T cell co-inhibition and type I interferon (IFN) response (Figure1 K-N), which indicating that the poor outcome might be caused by tumor-associated immune response dysfunction partly.

Conclusion: We constructed a robust immune gene prognostic model, which have a potential prognostic value for MDS patients and may provide evidence for personalized immunotherapy.

Figure

Disclosures

No relevant conflicts of interest to declare.
Retrospective Analysis of the Efficacy of Solvent/Detergent Treated Pooled Plasma As Compared to Fresh Frozen Plasma in Thrombotic Thrombocytopenic Purpura: A Single Center Experience
Authors: Klapheke Catherine A , Milito Chelsea , McRae Hannah L , Blumberg Neil , Refaai Majed A
Published: November 2020

Background : Plasma exchange is the primary treatment for acute episodes of Thrombotic Thrombocytopenic Purpura (TTP), a rare condition characterized by the formation of thromboses in small blood vessels, resulting in thrombocytopenia, hemolytic anemia and multi-organ failure. Plasma exchange replacement fluids in TTP include Fresh Frozen Plasma (FFP) and solvent/detergent treated pooled plasma (SDP), such as Octaplas. SDP is a virus-inactivated, pooled human plasma product with standardized plasma protein content, including ADAMTS13, that has been used in the treatment of TTP. This study evaluates the effectiveness of SDP as compared to FFP in the treatment of acute TTP episodes.

Study Design and Methods : A retrospective analysis was conducted comparing SDP- to FFP-treated patients with suspected acute TTP episodes as a primary admitting diagnosis between December 2014 and December 2019. A total of 16 patients were included in this study. The FFP group consisted of 9 patients (6M/3F, median age 44 years), three of whom had relapsed TTP. The SDP cohort had 7 patients (2M/5F, median age 38 years), one of whom had relapsed TTP. The primary outcomes measured included reported thromboembolic and major bleeding events. Secondary outcomes included number of plasma exchange procedures, adverse effects including neurological changes, transfusion of other blood products, ICU and hospital length of stay (LOS), and changes in laboratory values.

Results : There were no adverse transfusion reactions reported in either group. 12 bleeding events were reported from 6 patients in the FFP group and 8 bleeding events from 4 patients in the SDP group (p=0.397). No significant differences were detected in ICU stay, hospital LOS, number of plasma units transfused, daily percentage of change of laboratory values, or changes in neurological status.

Conclusions:These data confirm the previously reported efficacy of SDP for treating suspected/confirmed TTP.There were no significant differences in thromboembolic or bleeding events between patients who received FFP as compared to SDP. The non-significant differences observed in laboratory values throughout the duration of plasma exchange procedures as well as adverse reactions and clinical outcomes between both groups confirm the safety of SDP when used interchangeably with FFP for the treatment of TTP.

Table

Disclosures

Blumberg: CSL Behring: Consultancy. Refaai:CSL Behring: Consultancy; Octapharma: Research Funding; Instrumentation Laboratory: Research Funding; iLine Microsystems: Research Funding; Diagnostica Stago: Consultancy.
A Systemic Review on Efficacy of Ibrutinib-Based Regimens for the Treatment of Waldenstrom Macroglobulinemia
Authors: Khurshid Qasim , Dar Abdul Jabbar , Mirza Muhammad Ali , Ahmad Muhaddis Ejaz , Jaan Ali , Aamir Sobia , Abdullah Syed Maaz , Kashyap Richi , Khan Israr , Khan Waqas , Ali Sundas , Akbar Arshia , Javed Saad , Qamar Fazeel , Anwer Faiz
Published: November 2020

Introduction:

Ibrutinib works by inhibiting downstream signaling after the interaction between the mutated MYD88 (Leu265Pro) protein and BTK. In 2015, Ibrutinib, an inhibitor of Bruton's tyrosine kinase (BTK), was approved by the U.S Food and Drug Administration (FDA) for patients with symptomatic Waldenstrom Macroglobulinemia (WM).

Methods:

We performed a comprehensive literature search following PRISMA guidelines. Beginning with articles published after April 2015, we used databases like PubMed, Embase, Clinicaltrials.gov, Cochrane Library, and Web of Science. A total of 580 articles were identified initially, and after a detailed screening, we finalized 4 studies involving 248 WM patients.

Results:

The total number of patients who received Ibratunib based regimens were 248. The dose ranged from 140-560 mg. The overall response rate ranged from 90-92%, very good partial response ranged from 13-23%, and the partial response ranged from 36-75%.

1) Ibtrutunib +Rituximab:

In a study by Dimopoulos et al., (N=150), the overall response rate (ORR) was 92% vs 47% in control vs placebo respectively. The progression-free survival (PFS) of 30 months was 82% vs 28% respectively.

2) Ibrutinib alone:

In a study by Dimopoulos et al., with (N=31), ORR was 90%, a very good progression rate (VGPR) was 13 %, and the partial response (PR) was 58%. In a study by Treon et al., (N=63) ORR was observed in Treon et al. at 90.50%, VGPR was 15%, PR was 36%, no response (NR) was 57%, with overall survival (OS) was 24 months. In a study by Richard et al., (N=4), PR was noted at 75%.

Conclusion:

Ibtrutunib used for the treatment of WM showed an overall response rate of 92%. Infections, cytopenia, bleeding, hypertension and atrial fibrillation were major side effects reported. However, due to the recent FDA approval of the drug to be used for the patients of WM there is very limited studies on Ibrutinib. We recommend future randomized prospective trials to understand the efficacy and safety profile of Ibrutinib for WM treatment.

Disclosures

Anwer: Incyte, Seattle Genetics, Acetylon Pharmaceuticals, AbbVie Pharma, Astellas Pharma, Celegene, Millennium Pharmaceuticals.:Honoraria, Research Funding, Speakers Bureau.
Anlotinib Induced Apoptosis and Regulated the Chemosensitivity and Immune-Related Properties of Leukemia Stem Cells By Inhibiting JAK2-STAT3/5 Signaling
Authors: Liu Long , Jiang Long Yue , Xu Bing
Published: November 2020

Acute myeloid leukemia (AML) is derived from small populations of leukemia stem cells (LSCs) characterized by the self-renewal and chemoresistant properties. Residual LSCs after chemotherapy remain as the critical barriers to cure. Clearance of LSCs might rationally lead to an improvement of clinical outcome. Recently studies showed that JAK/STAT signaling play an important role in the self-renewal of AML-LSCs due to increased growth factor (GF) receptor expression such as c-kit, FLT3, CD123 and altered GF signaling by activating tyrosine kinases. Therefore, targeting such tyrosine kinases might be a strategy to eliminate LSCs. Anlotinib displayed its anti-tumor activity in lung cancer by targeting tyrosine kinase of VEGFR, FGFR, PDGFR and c-kit. However, whether anlotinib could inhibit the GF receptor-related tyrosine kinase overactivation and its downstream JAK-STAT signaling, and subsequently kill LSCs or regulate LSCs biology remains largely unknown.

To explore whether anlotinib could exert effective ani-LSCs activity, we treated LSC like cell lines (CD34+CD38-KG-1 and Kasumi-1) with anlotinib, and found anlotinib could effectively induce apoptosis of LSC-like cells in a dose- and time-dependent manner. Similar results were observed in primary CD34+CD38-AML LSCs; notably, anlotinib did not significantly kill normal CD34+ cells in vitro. Additionally, the anti-LSC activity of anlotinib was further confirmed in the xenograft mouse model by injection of Kasumi cells (LSC-like cell line) into irradiated female BALB/c nude mice.

To determine whether anlotinib could inhibit the over activation of the GF receptor-related tyrosine kinase, we performed western blot at 12h after anlotinib treatment when LSC-like cells did not showed significant apoptosis. As a result, anlotinib inhibit c-kit phosphorylation and JAK2 activation. Intriguingly, unlike JAK2 inhibitors, anlotinib could not only the inhibit phosphorylation of STAT3 and STAT5 but also downregulate their expression.

Chemoresistance and immune evasion were the key features of LSCs, JAK2-STAT3/5 signaling was reported to involved in chemoresistance by upregulating anti-apoptotic proteins such as Bcl-2 ,Mcl-1 and also involved in immune escape by inducing immune suppressive molecules such as PD-L1 ,TGF-β.Thus we evaluated Bcl-2 expression and found a significant decrease in LSC-likes cells after anlotinib treatment. Similarly, PD-L1 and TGF-β were also significantly downregulated after anlotinib treatment.

In conclusion, anlotinib not only displayed the effective anti-LSCs activity but also might regulate the chemoresistance and immune evasion of LSC by downregulating the anti-apoptotic proteins and suppressive molecules such as PD-L1, TGF-β respectively. Consequently, anlotinib might has the potential to contribute to a deeper clearance of LSCs by combining with chemotherapy or immunotherapy.

Disclosures

No relevant conflicts of interest to declare.
The Prognostic Impact of Dynamic Changes of Genetic Risk Stratification in Multiple Myeloma
Authors: Fan Huishou , Liu Jiahui , Yuan Chenglu , Mao Xue-Han , LI Xiaoqing , Du Xin , Li Zengjun , Hao Mu , Yan Yuting , Deng Shuhui , Du Chenxing , Li Chengwen , Zhao Jiawei , Yi Shuhua , Zou Dehui , Zhao Yaozhong , Zhan Fenghuang , Tai Yu-Tzu , Wang Jianxiang , Anderson Kenneth , Qiu Lugui , An Gang
Published: November 2020

Purpose: Most data on the prognostic impact of cytogenetic abnormalities come from the setting of newly diagnosed multiple myeloma, it remains unclear whether the dynamic changes of cytogenetic aberrations affect the prognostic evaluation in multiple myeloma.

Methods: We analyzed the prognostic impact of dynamic changes of cytogenetic abnormalities with a cohort consisting of 80 paired patients with consecutive cytogenetic data both at diagnosis and recurrence among 568 patients with newly diagnosed multiple myeloma.

Results : Three patterns of recurrence were established from 80 paired patients: Pattern A (40%) consisted of 32 patients without new cytogenetic abnormalities at the time of progression. Pattern B (15%) consisted of 12 patients harboring new standard risk (SR) cytogenetic aberrations. Pattern C (45%) consisted of 36 patients with new high-risk (HR) cytogenetic abnormalities. The median overall survival (mOS; P <0.001) and median progression-free survival (mPFS; P =0.013) differed significantly among three patterns. Four groups including 60 paired patients were further established according to genetic risk stratification changes. There were 15 patients who kept SR in group 1 (25%); 12 SR patients who evolved into HR group in group 2 (20%); 17 HR patients without new HR cytogenetic abnormalities in group 3 (28%); and 16 HR patients harboring new HR cytogenetic aberrations in group 4 (27%). The mOS (P=0.001) also differed significantly among these four groups.

Conclusions: Dynamic cytogenetic changes significantly affected prognostic evaluation in multiple myeloma. Patients harboring new HR cytogenetic abnormalities and escalated genetic risk stratification at relapse had worse outcomes.

Figure 1

Disclosures

Anderson: Bristol Myers Squibb: Membership on an entity's Board of Directors or advisory committees; Janssen: Membership on an entity's Board of Directors or advisory committees; Millenium-Takeda: Membership on an entity's Board of Directors or advisory committees; Sanofi-Aventis: Membership on an entity's Board of Directors or advisory committees; Oncopep and C4 Therapeutics.: Other: Scientific Founder of Oncopep and C4 Therapeutics.; Celgene: Membership on an entity's Board of Directors or advisory committees; Gilead: Membership on an entity's Board of Directors or advisory committees.
De-Escalation or Discontinuation of TKI in Patients with Chronic Myeloid Leukemia:a Multicentral,Open-Lable, Prospective Trial in China
Authors: Luo Jie , Tan Yaxian , Zhou Xuan , Xiaoli Liu , Liu Qifa , Xu Na
Published: November 2020

Aim:To observe and assess whether TKI can be de-escalated or discontinued without occurrence of molecular relapse in CML patients,who with continuous effective TKI over 5 years and maintain MR4.5(BCR-ABLIS≤0.0032%) at least 18 months.

Methods: In this study,we conducted half standard TKI dose or discontinued treatment in patients with CML who were aged 18 years and older and duration effective TKI treatment at least 5 years,duration MR4.5 monitored by quantitative polymerase chain reaction(Q-PCR) in recent 18 months.Blood concentration and immune function were also monitored.Molecular relapse was defined as loss of MMR(BCR-ABLIS>0.1%), these patients restore the original dose at once.

Results: 44 patients were enrolled between Jan 2016,and Jun 2019,35 in de-escalation group and 9 in discontinue group.In the de-escalation group,45.7% males,median age was 48(23-75) years,valid TKI therapy duration 84(60-204) months,and median sustained MR4.5 was 25(19-160) months.Follow-up 1 year,5(14.3%) of these 35 patients occurrence of molecular relapsed(3 before 6 months,one at month 8, one at month 9),all of them showed decreases in their BCR-ABL levels after reintroduction of original TKI dose.In the discontinue arm,55.6% males,the age ranged from 28 to 56,the median effective TKI treatment was 72(60-108) months and remained of MR4.5 was 36(18-72) months.Molecular recurrence-free events were 66.7% at 12 months,3 patients relapsed events occurred in discontinuous month 1,month 6 and month 12,respectively.All of them showed great reaction when treatment was restarted.Supervised patients blood concentration between standard treatment group and half-dose group,founded half-dose group was lower than standard group(1083.61±371.80ng/ml vs. 1622.28±771.36ng/ml ,P=0.000).The immune function of the relapse and non-relapse arm was observed,in relapsed group the proportion of CD3-/CD56+ cells was declined significantly(P=0.013).

Conclusions: Treatment-free remission(TFR) has become the main therapeutic target for CML patients. With 85.7% TFR in 12 months follow-up after de-escalation TKI treatment ,our study demonstrates that de-escalated TKI would suitable for Chinese CML patients who with a sustained deep molecular response.The vivo insufficient of drug concentration and the damaged of NK cells may cause the recurrence events.

Disclosures

No relevant conflicts of interest to declare.
A Case of CD5 Positive Hairy Cell Leukemia withBRAF V600EMutation and Lymph Node Involvement; Unique Case Report and Review of the Literature:
Authors: Javidiparsijani Sara , Choi Yun Kyong , Arbini Arnaldo A.
Published: November 2020

Objective:

Hairy cell leukemia (HCL) is distinct indolent neoplasm of small mature B cells accounting for 2% of the lymphoid leukemias. The classic immunophenotypic profile of HCL includes bright expression of B cell surface antigen and immunoglobulins and lack of both CD5 and CD10. The tumor cells are predominantly present in the spleen, peripheral blood and bone marrow. Aberrant expression of CD5 and lymph node involvement are very rare. A case of CD5(+) HCL with a documentedBRAF V600Emutation has not been previously reported. This case is also unique due to lymph node involvement by HCL. To our knowledge, this is the only case report of a documentedBRAF V600Emutated CD5(+) HCL along with lymph node involvement in the literature.

Method:

We are presenting a 45 years old man with multiple prior medical conditions including diabetes, COPD and interstitial pneomonitis with new onset of B-symptoms and neutropenia/monocytopenia. Peripheral blood flow cytometry revealed 21% CD5(+) CD10(-) monoclonal B cell population. Further workup revealed the neoplastic cells are positive for CD11c, CD103 and CD25. Bone marrow study revealed an extensive involvement of the marrow.By immunohistochemistry, the cells are also positive for CD5, CD20, BRAF and cyclin D1. Cytogenetic studies show that the cells are negative t(11;14) and molecular studies revealedBRAF V600Emutation. Reexamination of the prior wedge lung resection revealed intrathoracic lymph node involvement.

Results and conclusion:

The most common differential diagnoses of a CD5(+) B cell lymphoma involving the bone marrow is chronic lymphocytic leukemia (CLL) and mantle cell lymphoma. The key into the diagnosis of this case was marked monocytopenia that lead to addition of HCL antibody tubes despite the CD5 positivity and detection of CD25, CD103 and CD11c antigens in the leukemic cells. Bone marrow biopsy show extensive involvement by a CD5(+) cyclin D1(+) B cell lymphoma. The most important differential diagnosis in this case was mantle cell lymphoma. The diagnosis of HCL was confirmed by detection ofBRAF V600Emutation and absence of t (11;14). Retrospective examination of the intrathoracic lymph nodes show one lymph node involvement by hairy cell leukemia cells.

The patient was treated with Vemurafenib and his follow up flow cytometry revealed only 0.60% leukemic involvement. Incorporation of clinical data at the time of flow cytometric examination is essential to aid the hematopathologist in having a broader differential diagnoses and addition of appropriate tests that lead to the more accurate diagnosis and treatment.

Disclosures

No relevant conflicts of interest to declare.
Thrombopoietin Protects Neural Cells and Endothelial Cells from Apoptosis Via PI3K/AKT Pathway
Authors: Li Liang , Wang Junyan , Ye Jieyu , Yang Liuming , Chong Beng H , Yang Mo
Published: November 2020

Background: Thrombopoietin (TPO) is a hematopoietic growth factor that regulates the production of platelets and stimulates production and differentiation. The expression of TPO and TPO receptor (c-mpl) in the central nervous system (CNS) has been identified. However, the role of TPO in neural cells and endothelial cells were not clear.

Methods: C17.2 and human umbilical vein endothelial (HUVEC) cells were treated with CoCl2, TPO, or TPO + CoCl2. TPO was added into the culture medium 48 h before CoCl2 treatment. The cell viability and apoptosis of each group were tested by Cell Counter Kit 8 (CCK-8) assay and flow cytometry. The expression of Caspase-3 and mitochondrial membrane potential (MMP) were then determined by flow cytometry with Caspase-3-PE and JC-1. The effect of TPO in the PI3K/AKT pathway was detected by using Western blot.

Results: TPO has a dose-dependent effect on the growth of C17.2 cells. LY-294002 pretreatment suppressed the TPO-induced AKT activation and abolished the prosurvival effect of TPO. Via the Bcl-2/BAX signaling pathway, TPO exerted an anti-apoptotic effect by suppressing mitochondria membrane potentials. We also investigated the protective effect of TPO on human endothelial cells. The cell viability of HUVECs decreased gradually with the enhancement of CoCl2 at a gradient of chemical concentrations (r= -0.997). CoCl2 dramatically increased apoptosis of HUVECs, whereas pre-treatment with TPO rescued cell apoptosis induced by CoCl2 (P<0.01). Further investigation found that TPO decreased the expression of Caspase-3 and inhibited the reduction of MMP induced by CoCl2 (P<0.05). TPO increased the activation of PI3K/AKT pathway in HUVECs.

Conclusion: TPO has a protective effect against apoptosis of neural cells and endothelial cells through activating the PI3K/AKT pathway, thus decreasing the expression of apoptosis protease Caspase-3 and inhibiting the reduction of MMP.

Disclosures

No relevant conflicts of interest to declare.
An Azaindole-Based Small Molecule Hzx-02-059 Induces Methuosis in B-Cell Acute Lymphoblastic Leukemia through the PI3K/AKT Axis
Authors: Lu Zhen , Feng Liying , Chen Qinwei , Xu Bing
Published: November 2020

Methuosis is a newly defined nonapoptotic cell death characterized by the accumulation of large liquid-filled vacuoles in cytoplasm, which is distinct from necrosis, autophagy and apoptosis. A novel azaindole-based methuosis inducer, named HZX-02-059, displays cytotoxic effects to a series of cancer cellular models via blocking the role of PIKfyve. The discovery of HZX-02-059 sheds light on a novel therapeutic option for refractory cancer patients who have limited effective treatments. Despite significant advances in treatments, over 50% of adult patients with B-cell acute lymphoblastic leukemia (B-ALL) fail to achieve durable responses with current intensive chemotherapies, which highlight an urgent need of new therapeutic regimens for this patient population. Here, we applied HZX-02-059 to B-ALL cell lines and mice models to uncover its potential role and underlying mechanism.

Morphology changes of HZX-02-059-treated B-ALL cells were observed under the light microscope, and these treated cells were to certain extent collapsed into debris. Of note, higher concentration of this compound induced the accumulation of substantial larger vacuoles in cytoplasm. To assess the cytotoxicity of this compound, we performed cell counting kit-8 (CCK-8) and colony forming unit (CFU) assays. As expected, HZX-02-059 significantly inhibited cancer cell proliferation and reduced the capability of cell colony forming in a dose-dependent manner. Next, we carried out an EdU assay to determine the influence of HZX-02-059 on cell cycle distribution. As a result, B-ALL cells treated with HZX-02-059 were mainly blocked at G2/M phase. Western blotting was conducted to investigate the underlying molecular basis of HZX-02-059 in B-ALL. To the end, we found that HZX-02-059 negatively regulated the proteins involved in the PI3K/AKT pathway. Also, this drug decreased the expression of the pathway downstream transcription factors NF-ǐB and c-Myc, suggesting that the dysregulation of the PI3K/AKT axis is a potential mechanism of action of HZX-02-059 against B-ALL.

To further evaluate itsin vivotherapeutic effect, B-ALL patient-derived xenograft (PDX) mice were established and administrated with HZX-02-059. The result revealed a survival benefit of the treatment as compared with the control, which grants this molecule great potential for clinical usage. Thus, our results suggest that HZX-02-059 is a potent anti-BALL reagent in preclinical models that warrant further investigation of this drug as an alternative therapeutic strategy for B-ALL patients who are resistant to conventional therapies.

Disclosures

No relevant conflicts of interest to declare.
Clinical Trial to Evaluate an Approved ITP Therapy Targeting Spleen Tyrosine Kinase (SYK) for Prevention and Treatment of COVID-19 Related Complications
Authors: Dummer Wolfgang , Markovtsov Vadim V. , Tong Sandra , Masuda Esteban
Published: November 2020

Fostamatinib is a potent SYK inhibitor that targets activation of both the innate and acquired immune systems caused by tissue damage or pathogen signals via C-type lectin receptors (CLR) as well as by antibody-antigen immune complexes via Fc receptors (Figure 1). SYK is also a key modulator of pathways involved in thrombosis formation. Fostamatinib is FDA-approved for chronic immune thrombocytopenia (ITP) and has been evaluated in over 4000 patients for treatment of allergic, autoimmune, and neoplastic disorders. Fostamatinib is well tolerated, with a consistent safety profile across a number of diseases. In rheumatoid arthritis patients, fostamatinib reduced the plasma levels of IL-6.1 The active metabolite (R406) was protective in mouse models of LPS-induced acute lung injury (ALI)2 and LPS- or antibody-induced acute kidney injury (AKI).3 SYK inhibition decreased the incidence of thrombosis in mouse models of thromboembolism,4 and the incidence of thromboembolic events was decreased in patients treated with fostamatinib compared with other treatments for ITP. These immunomodulatory effects suggest that SYK inhibition represents a new therapeutic strategy for the treatment of COVID-19, which led to design of a clinical study of fostamatinib in hospitalized COVID-19 patients.

COVID-19 patients are at risk for a potentially fatal acute respiratory distress syndrome (ARDS), cytokine storm, severe systemic capillary leak syndrome, thromboembolic events, and multi-organ dysfunction/failure. SYK is a master regulator of signal transduction pathways implicated in these COVID-19 associated complications, which involves hyperactivation of both innate and acquired immune systems. Targeting SYK could prevent the cytokine storm, downstream activation of endothelial cells and platelets, and influx of neutrophils and monocytes into lungs or kidney leading to ALI and AKI, respectively.

Methods: A randomized, double-blind, placebo-controlled, multi-center, Phase 3 study will evaluate the efficacy and safety of fostamatinib in adult, hospitalized, high-risk COVID-19 patients. Patients will be treated with fostamatinib 150mg BID for up to 28 days plus standard of care. Improvement in clinical status will be assessed using the 8-point ordinal scale.

References

1. Weinblatt ME et al.Arth Rheum2008;58:3309-18

2. Nadeem A et al.Int Immunopharm2019;68:39-47

3. Al-Harbi NO, et al.Biochimie2019;158: 102-10

4. Van Eeuwijk JMM et al.Arterioscler Thromb Vasc Biol2016;36:1247-53

Results and Conclusions: Trial in Progress

Disclosures

Dummer: Rigel:Current Employment, Current equity holder in publicly-traded company.Markovtsov:Rigel:Current Employment, Current equity holder in publicly-traded company.Tong:Rigel:Current Employment, Current equity holder in publicly-traded company.Masuda:Rigel:Current Employment, Current equity holder in publicly-traded company.
A Systematic Review of Evidence Supporting the Use of Autologous Cell Vaccines in the Treatment of Hematological Malignancies
Authors: Bastin Donald , Khan Sarwat , Montroy Joshua , Kennedy Michael A. , Forbes Nicole , Martel Andre B , Baker Laura , Gresham Louise , Boucher Dominique , Wong Boaz , Diallo Jean-Simon , Fergusson Dean , Lalu Manoj , Auer Rebecca C , Kekre Natasha
Published: November 2020

By presenting a patient's own tumor antigens to their immune system, autologous cancer cell vaccines can drive a robust polyclonal adaptive response. In this systematic review and meta-analysis, we investigated the safety and efficacy of these vaccines administered to patients with hematologic malignancies. Our primary outcomes of interest were safety and clinical response, with secondary outcomes including overall, disease-free and progression-free survival, relapse rate, correlative immune assays and health-quality related metrics. We identified 14 studies with 332 patients enrolled, of which 200 were ultimately treated with at least one dose of the vaccine. While we identified both patient-related and technical issues that might limit the feasibility of these trials, very few serious adverse events (AEs) were reported overall, with only 31.5% of patients suffering any AEs. Grade II or lower AE was observed in 10 (71.4%) of the 14 studies. Of the 3 (21.4%) reporting grade IV AEs, two observed the AE in one patient only, and one reported a 20% incidence of grade II-IV AEs. Of 58 evaluable patients, the complete response rate was 21% [95% CI, 10%-38%)] and overall response rate was 36% [95% CI, 24%-49%]. Analysis of individual patient level data (n=50) revealed a 5-year overall survival of 68.7% (SE 7.1%) and disease-free survival of 67.4% (SE 8.1%). Despite the clear safety and a signal towards efficacy, our review has identified several factors limiting administration of these vaccines, which should be considered when developing future clinical trials.

PROSPERO registration number CRD42019140187

Disclosures

Diallo: Virica Biotech: Other: Owner and Executive. Auer:Imugene: Other: Scientific Advisory Board. Kekre:Gilead: Consultancy, Honoraria; Novartis: Consultancy, Honoraria; Celgene: Consultancy, Honoraria.
CD56briCD27+CD11b+/- NK Cells Mediated Remission of Agvhd Via Immunoregulation
Authors: Ni Ming , Zhao Peng , Kang Qian , Zhao Mei , Liu Min , Zhang Fengqi , Huang Shisi , Ma Dan , Zhang Yaming , Zhao Jiangyuan , Wang Jishi
Published: November 2020

Background: In our previous study CD56briCD27+CD11b+/- NK cells show the potential of the remission of aGvHD, while the mechanism behind it is still unknown. Here, three hypothesis have been proposed according to further pilot experiment and literature.

Method: 74 samples of patients after allo-HSCT were collected in the affiliated hospital of Guizhou medical university. Immunomonitoring with respect to the phenotype and function of CD56briCD27+CD11b+/- NK cells was performed on rested peripheral blood mononuclear cells (PBMCs) using multiparametric flow cytometry. Furthermore, the western-blot has been performed to determine the expression of HO-1 protein in sorted CD56briCD27+CD11b+/- NK cells as well.

Result: (1) Our result shows a positive correlation between CD56briCD27+CD11b+/- NK cells and Th22, Tfh as well as Th1 like Tfh cells. Interestingly, a dramatic higher expression of homing marker CD62L has been observed on CD56briCD27+CD11b+/- NK cells. Thus we hypothesis that the CD56briCD27+CD11b+/- NK cells may migrate into the lymphoid tissues and regulate the differentiation of naïve T cells. (2) On the other hand, an elevated percentage of NKG2D has been found on CD56briCD27+CD11b+/- NK cells in patients with aGvHD. Meanwhile a high expression of Fas and exhaustion markers have been observed on CD4+ and CD8+ T cells. These results suggest that T cells may be eliminated by CD56briCD27+CD11b+/- NK cells. (3) Besides, our work displays that the expression of Heme Oxygenase-1 (HO-1) in CD56briCD27+CD11b+/- NK cells positively correlated with the secretion of IL-10. Compared with the patients with aGvHD, a higher expression of HO-1 and IL-10 in CD56briCD27+CD11b+/- NK cells has been found in patients without aGvHD at day 30 after allo-HSCT .

Conclusion: According to these findings, the CD56briCD27+CD11b+/- NK cells may result in the remission of aGvHD via immunoregulation by regulation of T cells and secretion of IL-10.

Disclosures

No relevant conflicts of interest to declare.
A Unique Case of CSF3R-Mutated Chronic Neutrophilic Leukemia with Thrombocytosis
Authors: Tizro Parastou , Yacur Melissa , Liu Min-Ling , Nava Victor E. , Aggarwal Anita
Published: November 2020

Chronic neutrophilic leukemia (CNL) is a well-recognized, extremely rare myeloproliferative neoplasm (MPN) with only ~200 reported cases until 2017. The current WHO diagnostic criteria include leukocytosis of ≥ 25 x 109/L with ≥ 80% neutrophils/bands, < 10% circulating neutrophil precursors and <1% blasts (<5% bone marrow blasts). Exclusion criteria includes dysplasia, or clinical/molecular diagnosis of other MPN. The WHO classification was updated in 2016 to include CSF3R T618I and other activating mutations in CSF3R as diagnostic. SETBP1 and ASXL1 mutations are frequent but unspecific in CNL and appear to be of prognostic significance. Hepatosplenomegaly and a hypercellular bone marrow with marked granulocytic hyperplasia are common in CNL, but thrombocytosis has not been reported previously.

We report a 72-year-old man with past medical history of hypertension and hepatitis C status post-treatment, who presented with WBC 38.4k/cmm, Hgb 6.3g/dl, MCV 102 fl and platelet count of 2601k/cmm. Reactive etiology of thrombocytosis was ruled out. Peripheral blood smear revealed rare blasts. Bone marrow biopsy was hypercellular (60-80%) with M:E ratio of 10:1, granulocytic and megakaryocytic hyperplasia, clustering of dyspoietic megakaryocytes, no erythroid or myeloid dysplasia and minimal patchy reticulin fibrosis suggestive of primary MPN. Chromosome analysis showed normal karyotype of 46 XY. Molecular studies were negative for BCR-ABL, JAK-2 V617F, CALR and MPL, Next Generation Sequencing revealed mutations in the CSF3R gene (Q781* and T618I) and concurrent inactivating ASXL1-G646fs*12 and activating U2AF1-Q157P mutations. These findings were compatible with a diagnosis of CNL.

We describe an unusual case of CNL confirmed by CSF3R gene mutations (T618I and Q781*) with significant thrombocytosis and without splenomegaly. Although the T618I mutation has commonly been reported in CNL, to the best of our knowledge, the Q781* mutation is novel in CNL, and has only been reported before in severe congenital neutropenia (SCN). Association of Q781* mutation in CSF3R gene with significant thrombocytosis in our patient needs further investigation.

Disclosures

No relevant conflicts of interest to declare.
Nicotinamide (NA) Treatment Improves Response to G-CSF in Severe Congenital Neutropenia (SCN) Patients
Authors: Deordieva Ekaterina , Skokowa Julia , Shcherbina Anna , Novichkova Galina , Maschan Alexey , Welte Karl
Published: November 2020

Background: SCN is an inherited bone marrow failure syndrome with markedly reduced granulocyte numbers and functions that often manifests with life-threatening bacterial infections. G-CSF treatment is the primary therapeutic approach, yet some SCN patients require high doses of G-CSF which increases the risk of acute myeloid leukemia development, or do not respond to GCSF at al. We have demonstrated that nicotinamide (NA) treatment increases granulocyte numbers in healthy volunteers.

Aim: To asses the effect of NA treatment in SCN and cyclic neutropenia (CyN) patients.

Methods: 15 SCN and three CyN patients ages 0.5 to 31 (M-5.47) years were orally treated with 20 mg/kg/day of NA in addition to G-CSF therapy. G-CSF dose varied between 0.6 and 50.8 µg/kg/day before start of NA treatment.

Results: After three months of NA treatment, absolute neutrophil counts (ANC) increased from median 0.80 x 109/l (Q1 0.61, Q3 1.29 x 109/l) to 1.77 x 109/l (Q1 1.49, Q3 2.33 x 109/l) (p < 0.001). This allowed G-CSF dose reduction from 15 µg/kg/day (Q1 5, Q3 25 µg/kg/day) to 10 µg/kg/day (Q1 1.77, Q3 15.16 µg/kg/day) in 17 patients, and complete G-CSF tapering in one patient. After one year of observations, ANC were 1.50 x 109/l (Q1 0.99, Q3 2.86 x 109/l), median increase of ANC from start of therapy was 0.89 x 109/l fold (Q1 0.39, Q3 1.57; р = 0.004). NA was well tolerated, without severe adverse events. We also observed a marked reduction of the frequency and severity of bacterial infections upon combination of NA and G-CSF.

Conclusions: In our group of SCN/CyN patients, NA treatment led to increased neutrophil counts and decreased required G-CSF doses with continuous clinical and laboratory responses. We also observed a reduction of the frequency and severity of bacterial infections. Therefore, the use of NA in combination with a reduced dose of G-CSF for the treatment of SCN and CyN patients is promising and should be further investigated in a larger cohort of patients.

Disclosures

No relevant conflicts of interest to declare.
Differential Use of Rap1 Effectors for Integrin Activation in Platelets and Lymphocytes
Authors: Lagarrigue Frederic , Sun Hao , Gingras Alexandre R , Ginsberg Mark H
Published: November 2020

Rap1 is a major convergence point of the platelet and lymphocyte signaling pathways that result in talin1 binding to the integrin β-cytoplasmic domain and consequent integrin activation, effective hemostasis and immune functions. The nature of the connection between Rap1 and talin1 in integrin activation is an important remaining gap in our understanding of this process. Deletion of RIAM in T cells partially suppressed αLβ2 and α4β7 integrin activation but did not block activation of α4β1. In sharp contrast, deletion of both Rap1a and Rap1b isoforms or deletion of talin1 profoundly suppressed activation of α4β1 in addition to the other two integrins. The loss of homing of T cells to peripheral lymphoid organs was correlated with the loss of integrin activation in these mutant T cells. We recently identified two Rap1 binding sites in talin1 F0 and F1 domains and generated mice bearing point mutations, which block Rap1 binding to both talin1 F0 and F1 domains (R35E,R118E). In platelets, which lack endogenous RIAM, a talin1 (R35E,R118E) mutation suppresses activation of integrin αIIbβ3 and platelet aggregation to a similar extent to that observed in Rap1a,b null platelets. In contrast, talin1 (R35E,R118E)-expressing T cells manifest only partial loss of integrin activation, whereas RIAM null T cells expressing talin1 (R35E,R118E) exhibit a much more profound defect. Importantly, overexpression of RIAM can completely rescue the integrin activation defect in talin1 (R35E,R118E)-expressing T cells indicating that Rap1 binding to RIAM and talin1 are partially redundant in T cells. These findings reveal that although both Rap1 and talin1 are essential for integrin action in blood cells, the connections between them vary in different blood cells and that the final steps in integrin activation and blood cell function can be manipulated in a cell-type specific manner.

Disclosures

No relevant conflicts of interest to declare.
Venous Thromboembolic Disease in Children Associated with COVID-19
Authors: Narang Shalu , Morparia Kavita , Sharma Prachi
Published: November 2020

Venous thromboembolism (VTE) in children is rare and VTE in association with antiphospholipid antibodies (lupus anticoagulant, cardiolipin antibodies and beta-2 glycoprotein I) is even rarer. The development of these antibodies can be due to an acute infection or an autoimmune disease. Recently, these antibodies have been noted to be present in conjunction with SARS-CoV-2 in adult patients with VTE.

We report 5 cases of VTE associated with presence of antiphospholipid antibodies in children over a short period during the coronavirus 19 pandemic. One child had SARS-CoV-2 PCR positive on nasopharyngeal swab while 2 had positive antibodies (IgM) to SARS-CoV-2. One adolescent female had features of Multi-Inflammatory System in Children (MIS-C) a few weeks before presenting with multiple pulmonary emboli and DVT. Thrombotic events included DVT of lower extremity (3 patients), multiple PE (2 patients), orbital vein thrombosis (1 patient) and upper extremity DVT (1 patient). Two of these patients had both DVT and PE. All patients had one or more antiphospholipid antibody present. Two adolescent females had new onset of systemic lupus erythematous. All patients were treated with systemic anticoagulation with heparin (initially with unfractionated heparin if clinically unstable and later changed to low molecular weight heparin).

We report a cluster of adolescents with unusual thrombotic complications with antiphospholipid antibody positivity during the coronavirus 19 pandemic, 3 with positive virologic criteria, one with delayed presentation after MIS-C like features and one with close family member exposure. We hypothesize that VTE in children is likely due to development of antiphospholipid antibodies when associated with SARS-CoV-2 infection.

Disclosures

No relevant conflicts of interest to declare.