Synergistic suppression of the PI3K inhibitor CAL-101 with bortezomib on mantle cell lymphoma growth

Objective: To investigate the effects of CAL-101, particularly when combined with bortezomib(BTZ) on mantle cell lymphoma(MCL) cells, and to explore its relative mechanisms.Methods: MTT assay was applied to detect the inhibitory effects of different concentrations of CAL-101. MCL cells were divided into four groups: control group, CAL-101 group, BTZ group, and CAL-101/BTZ group. The expression of PI3K-p110σ, AKT, ERK, p-AKT and p-ERK were detected by Western blot. The apoptosis rates of CAL-101 group, BTZ group, and combination group were detected by flow cytometry. The location changes of nuclear factor kappa-B(NF-κB) of 4 groups was investigated by NF-κB Kit exploring. Western blot was applied to detect the levels of caspase-3 and the phosphorylation of AKT in different groups. Results: CAL-101 dose- and time-dependently induced reduction in MCL cell viability. CAL-101 combined with BTZ enhanced the reduction in cell viability and apoptosis. Western blot analysis showed that CAL-101 significantly blocked the PI3K/AKT and ERK signaling pathway in MCL cells. The combination therapy contributed to the inactivation of NF-κB and AKT in MCL cell lines. However, cleaved caspase-3 was up-regulated after combined treatment. Conclusion: Our study showed that PI3K/p110σ is a novel therapeutic target in MCL, and the underlying mechanism could be the blocking of the PI3K/AKT and ERK signaling pathways. These findings provided a basis for clinical evaluation of CAL-101 and a rationale for its application in combination therapy, particularly with BTZ.

再生障碍性贫血病因学的新探索:异常免疫诱导骨髓间充质干细胞的过度脂肪化(英文)

再生障碍性贫血(简称再障)是一种以血细胞减少为表现的疾病。骨髓脂肪过多且造血前体细胞减少是其显著骨髓病理学特征。本文解读和分析了与再障和脂肪细胞相关的论著和关键综述,旨在对骨髓脂肪细胞在再障发病中的作用进行初步探索。这些文献提示,再障相关性毒素和毒性药物,一般也具有诱导骨髓间充质干细胞向脂肪细胞分化的作用。再障的免疫治疗反应时间要远远长于典型免疫相关性血细胞减少症,表明异常免疫的靶细胞不仅仅是造血干/祖细胞,骨髓间充质干细胞也可能也受累。免疫抑制剂和雄激素治疗再障有效,二者也具有抑制骨髓间充质干细胞向脂肪细胞分化的作用。脂肪细胞分泌的很多细胞因子均有抑制骨髓造血作用,包括肿瘤坏死因子(TNF-α),白细胞介素6(IL-6)和干扰素γ(IFN-γ)。动物模型也证实,骨髓过度脂肪化可抑制造血。另外,获得性再生障碍性贫血一般被视作一种免疫介导的疾病,包括免疫细胞和分子介导造血干/祖细胞破坏导致骨髓造血衰竭。综上,骨髓内间充质干细胞向脂肪细胞过度分化可能是再障发病的新的病理学机制之一。

人原发性胃恶性淋巴瘤裸鼠原位移植肝转移模型的建立(英文)

Background and Object ive:In recent years,incidence and mortality of lymphoma are markedly increasing worldwide.However,the pathogenesis and mechanism of invasion and metastasis for lymphoma are not yet fully clarified.It is mainly due to the lack of ideal animal models,which can precisely simulate the invasion and metastasis of lymphoma in the human body.So,it is very necessary to establish a highly metastatic nude mouse model of human lymphoma.This study developed a livermetastatic model of primary gastric lymphoma in nude mice by using orthotopic surgical implantation of histologically intact patient specimens into the corresponding organs of the recipient small animals.Met hods:A histologically intact fragment of liver metastasis derived from a surgical specimen of a patient with primary gastric lymphoma was implanted into the submucosa of the stomach in nude mice.Tumorigenicity,invasion,metastasis,morphologic characteristics(via light microscopy,electron microscopy,and immunohistochemistry),karyotype analysis,and DNA content of the orthotopically transplanted tumors were studied.Result s:An orthotopic liver metastatic model of human primary gastric lymphoma in nude mice(termed HGBL-0304) was successfully established.The histopathology of the transplanted tumors showed primary gastric diffuse large B-cell lymphoma.CD19,CD20,CD22,and CD79α were positive,but CD3 and CD7 were negative.The serum level of lactate dehydrogenase(LDH) was elevated [(1010.56 ± 200.85) U/L].The number of chromosomes ranged from 75 to 89.The DNA index(DI) was 1.45 ± 0.25(that is,heteroploid).So far,the HGBL-0304 model has been passed on for 45 generations of nude mice.A total of 263 nude mice were used for the transplantation.Both the growth and resuscitation rates of liquid nitrogen cryopreservation of the transplanted tumors were 100%.The transplanted tumors autonomically invasively grew and damaged a whole layer in the stomach of nude mice.The metastasis rates of liver,spleen,lymph node,and peritoneal seeding were 100%,94.3%,62.6%,and 43.5%,respectively.Conclusions:The study successfully establishes an orthotopic liver metastatic model of human primary gastric lymphoma in nude mice.The HGBL-0304 model can completely simulate the natural clinical process of primary gastric lymphoma and provides an ideal animal model for the research on the biology of metastasis and antimetastatic experimental therapies of primary gastric lymphoma.

Rhabdomyolysis induced by simvastatin-diltiazem interaction in unrecognized hypothyriodism

Simvastatin ，一个 3-hydroxy-3-methylglutaryl 辅酶 A reductase 禁止者，广泛地与 hypercholesteremia 被规定到病人，它的肌肉发达的毒性是广泛地， simvastatin 的 reported.The 新陈代谢由与 simvastatin.Diltiazem 交往在临床的事件取决于细胞色素 P450 3A4 ( CYP3A4 )和 CYP3A4 罐头结果的禁止者的 enzymic 活动是 CYP3A4 的一个中等禁止者，它被知道增加我们报导的 simvastatin.Here 的浆液集中有 unrecogniz

NADPH Oxidase Accounts for Changes in Cerebrovascular Redox Status in Hindlimb Unweighting Rats

Objective The roles of cerebrovascular oxidative stress in vascular functional remodeling have been described in hindlimb-unweighting （HU） rats. However, the underlying mechanism remains to be established. Methods We investigated the generation of vascular reactive oxygen species （ROS）, Nox2/Nox4 protein and mRNA levels, NADPH oxidase activity, and manganese superoxide dismutase （MnSOD） and glutathione peroxidase-1 （GPx-1） mRNA levels in cerebral and mesenteric smooth muscle cells （VSMCs） of HU rats. Results ROS production increased in cerebral but not in mesenteric VSMCs of HU rats compared with those in control rats. Nox2 and Nox4 protein and mRNA levels were increased significantly but MnSOD/GPx-1 mRNA levels decreased in HU rat cerebral arteries but not in mesenteric arteries. NADPH oxidases were activated significantly more in cerebral but not in mesenteric arteries of HU rats. NADPH oxidase inhibition with apocynin attenuated cerebrovascular ROS production and partially restored Nox2/Nox4 protein and mRNA levels, NADPH oxidase activity, and MnSOD/GPx-1 mRNA levels in cerebral VSMCs of HU rats. Conclusion These results suggest that vascular NADPH oxidases regulate cerebrovascular redox status and participate in vascular oxidative stress injury during simulated microgravity.

Imatinib-induced decompensated heart failure in an elderly patient with chronic myeloid leukemia: case report and literature review

Because it is safe and well tolerated, imatinib is a standard first-line therapy for chronic myeloid leukemia (CML). Although there have been sporadic reports of imatinib-induced cardiotoxicity, including left ventricle (LV) dysfunction and heart failure, the evidence for it is contradictory. Here, we reported a case of an 88-year-old male patient with CML developed decompensated heart failure following imatinib therapy. Four days after the initiation of imatinib, the patient developed orthopnea, edema and a pleural effusion accompanied by abdominal distension, nausea and vomiting. The chest X-ray film showed an enlarged cardiac profile. The echocardiogram demonstrated a decreased LV ejection fraction and enlarged left-side cardiac chambers. B-type natriuretic peptide concentrations were markedly increased. The patient recovered soon after the withdrawal of imatinib and introduction of comprehensive therapy for heart failure. Imatinib-induced cardiotoxicity in elderly patients is a potentially serious complication thatmerits further evaluation.

Phase I study of chimeric antigen receptor modified T cells in treating HER2-positive advanced biliary tract cancers and pancreatic cancers

This phase I clinical trial （NCT01935843） is to evaluate the safety, feasibility, and activity of chimeric antigen receptor-engineered T cell （CART） immunotherapy tar- geting human epidermal growth factor receptor 2 （HER2） in patients with advanced biliary tract cancers （BTCs） and pancreatic cancers （PCs）. Eligible patients with HER2-positive （〉50%） BTCs and PCs were enrolled in the trial. Well cultured CART-HER2 cells were infused following the conditioning treatment composed of nab- paclitaxel （100-200 mg/m2） and cyclophosphamide （15-35 mglkg）. CAR transgene copy number in the peripheral blood was serially measured to monitor the expansion and persistence of CART-HER2 cells in vivo. Eleven enrolled patients received 1 to 2-cycle CART- HER2 cell infusion （median CAR＋ T cell 2.1× 10^6/kg）. The conditioning treatment resulted in mild-to-moderate fatigue, nausea/vomiting, myalgialarthralgia, and lym- phopenia. Except one grade-3 acute febrile syndrome and one abnormal elevation of transaminase （〉9 ULN）, adverse events related to the infusion of CART-HER2 cells were mild-to-moderate. Post-infusion toxicities included one case of reversible severe upper gastroin- testinal hemorrhage which occurred in a patient with gastric antrum invaded by metastasis 11 days after the CART-HER2 cell infusion, and 2 cases of grade 1-2delayed fever, accompanied by the release of C-reactive protein and interleukin-6. All patients were evaluable for assessment of clinical response, among which 1 obtained a 4.5-months partial response and 5 achieved stable disease. The median progression free survival was 4.8 months （range, 1.5-8.3 months）. Finally, data from this study demonstrated the safety and feasibility of CART-HER2 immunotherapy, and showed encourag- ing signals of clinical activity.

An analytical biomarker for treatment of patients with recurrent B-ALL after remission induced by infusion of anti-CD19 chimeric antigen receptor T(CAR-T) cells

Anti-CD19 chimeric antigen receptor-modified T(CAR-T-19) cells have emerged as a powerful targeted immunotherapy for B-cell lineage acute lymphoblastic leukemia with a remarkable clinical response in recent trials. Nonetheless, few data are available on the subsequent clinical monitoring and treatment of the patients, especially those with disease recurrence after CAR-T-19 cell infusion. Here, we analyzed three patients who survived after our phase I clinical trial and who were studied by means of biomarkers reflecting persistence of CAR-T-19 cells in vivo and predictive factors directing further treatment. One patient achieved 9-week sustained complete remission and subsequently received an allogeneic hematopoietic stem cell transplant. Another patient who showed relapse after 20 weeks without detectable leukemia in the cerebrospinal fluid after CAR-T-19 cell treatment was able to achieve a morphological remission under the influence of stand-alone low-dose chemotherapeutic agents. The third patient gradually developed extensive extramedullary involvement in tissues with scarce immune-cell infiltration during a long period of hematopoietic remission after CAR-T-19 cell therapy. Long-term and discontinuous increases in serum cytokines(mainly interleukin 6 and C-reactive protein) were identified in two patients(Nos. 1 and 6) even though only a low copy number of CAR molecules could be detected in their peripheral blood. This finding was suggestive of persistent functional activity of CAR-T-19 cells. Combined analyses of laboratory biomarkers with their clinical manifestations before and after salvage treatment showed that the persistent immunosurveillance mediated by CAR-T-19 cells would inevitably potentiate the leukemia-killing effectiveness of subsequent chemotherapy in patients who showed relapse after CAR-T-19-induced remission.

Treatment of CD20-directed Chimeric Antigen Receptor-modified T cells in patients with relapsed or refractory B-cell non-Hodgkin lymphoma:an early phase IIa trial report

Patients with relapsed or refractory non-Hodgkin lymphoma have a dismal prognosis.Chimeric Antigen Receptor(CAR)-modified T cells(CART cells)that targeted CD20 were effective in a phase I clinical trial for patients with advanced B-cell lymphomas.We performed a phase IIa trial to further assess the safety and efficacy of administering autologous anti-CD20 CART(CART-20)cells to patients with refractory or relapsed CD20^(+)B-cell lymphoma.Eleven patients were enrolled,and seven patients underwent cytoreductive chemotherapy to debulk the tumors and deplete the lymphocytes before receiving T-cell infusions.The overall objective response rate was 9 of 11(81.8%),with 6 complete remissions(CRs)and 3 partial remissions;no severe toxicity was observed.The median progression-free survival lasted for 46 months,and 1 patient had a 27-month continuous CR.A significant inverse correlation between the levels of the CAR gene and disease recurrence or progression was observed.Clinically,the lesions in special sites,specifically the spleen and testicle,were refractory to CART-20 treatment.Collectively,these results together with our data from phase I strongly demonstrated the feasibility and efficacy of CART-20 treatment in lymphomas and suggest large-scale patient recruitment in a future study.This study was registered at www.clinicaltrials.org as NCT01735604.

Angiogenesis related gene expression profiles of EA.hy926 cells induced by irbesartan： a possible novel therapeutic approach

Background Angiogenesis occurs commonly in various physiological and pathological processes. Improving blood supply through promoting angiogenesis is a novel approach for treating ischemic diseases. Angiotensin II type 1 receptor blockers （ARBs） dominate the management of hypertension, but evidence of their role in angiogenesis is contradictory. Here we explored the angiogenic effects of ARBs through characterizing gene expression of the human umbilical vein endothelial cell line EA.hy926 exposed to irbesartan. Methods The human umbilical vein endothelial cell line EA.hy926 was grown for 72 hours after treatment with different concentrations of irbesartan. The cell proliferative capacity was assessed by CCK8 assay at 24, 48 and 72 hours. Gene expression levels in EA.hy926 cells responding to irbesartan were measured under optimal proliferation conditions by microarray analysis using Affymetrix U133 plus 2.0. The differential expression of genes involved in angiogenesis was identified through cluster analysis of the resulting microarray data. Quantitative RT-PCR and Western blotting analyses were used to validate differential gene expression related to the angiogenesis process. Results In the 104, 105, 106 mol/L treatment groups, cell proliferation studies revealed significantly increased proliferation in EA.hy926 cells after 24 hours of irbesartan treatment. However, after 48 and 72 hours of treatment with different concentrations of irbesartan, there was no significant difference in cell proliferation observed in any treatment group. We selected the group stimulated with irbersartan at a concentration of 10.6 mol/L for microarray experiments. Statistical analysis of the microarray data resulted in the identification of 56 gene transcripts whose expression patterns were significantly correlated, negatively or positively, with irbesartan treatment. Cluster analysis showed that these genes were involved in angiogenesis, extracellular stimulus, binding reactions and skeletal system morphogenesis. Of these 56 genes we identified seven genes （VEGF, KDR, PTGS2, PLXND1, ROB04, LM02, and COL5A1） involved in the angiogenesis process, qRT-PCR analysis of these genes confirmed the microarray results. Protein expression of three VEGF pathway genes （VEGF, KDR, and PTGS2） was further confirmed by Western blotting. Conclusions Our study showed that irbesartan may induce angiogenic effects in vascular endothelial cells. It suggested that the mechanism of angiogenic effects of ARBs might be attributed to the signaling cascade from angiotensin receptors in the VEGF pathway. It also provided evidence indicating that ARBs could be used as a novel therapeutic approach to treat chronic ischemic heart disease as well as anti-hypertensive agents.

Efficiency and side effects of anti-CD38 CAR T cells in an adult patient with relapsed B-ALL after failure of bi-specific CD19/CD22 CAR T cell treatment

(CAR)T cells and immune checkpoint blockade therapies has shown remarkable outcomes in the field of cancer treatment.1,2 In particular,great success has been achieved in B cell acute lymphocytic leukemia(B-ALL)treated with anti-CD19 CAR T cells.2 However,the loss,mutation,and reduced expression of CD19 are crucial causes for the failure of CAR T cell treatment in leukemia patients.3,4 CD38,a 45-kDa type II glycoprotein,is detectable in BALL,and several studies have indicated the efficacy and safety of CD38 monoclonal antibodies in clinical applications,suggesting that CD38 represents a suitable therapeutic target for the treatment of B-ALL.5,6 In addition,we developed a CAR against CD38,and our preclinical study indicated that anti-CD38 CAR T(CAR T-38)cells could specifically lyse CD38-positive tumor cells.7 Herein,we present the anti-leukemia ability of CAR T-38 cells in an adult relapsed B-ALL patient after failure of bi-specific CD19/CD22 CAR T cell treatment,and this administration of CAR T-38 cells was accompanied by the occurrence of target-mediated toxicities and severe and uncontrollable cytokine release syndrome(CRS).

Long-term safety and efficacy of CART-20 cells in patients with refractory or relapsed B-cell non-Hodgkin lymphoma:5-years follow-up results of the phase I and IIa trials

For refractory or relapsed(r/r)B-cell non-Hodgkin lymphoma(NHL),the response rates to conventional salvage chemotherapy are 27–44%.1 Chimeric antigen receptors(CARs)efficiently redirect T-cell specificity and cytotoxicity to cells expressing the targeted Ag in an HLA-independent manner.2 The early phase clinical trials of CART cells for(r/r)B-cell NHL have demonstrated promising results..