Reconstitution of the Multiple Myeloma Microenvironment Following Lymphodepletion with BCMA CAR-T Therapy.

PURPOSE: To investigate the remodeling of multiple myeloma (MM) microenvironment after BCMA-targeted chimeric antigen receptor T cell (CAR-T) therapy. EXPERIMENTAL DESIGN: We performed single-cell RNA sequencing (scRNA-seq) on paired bone marrow specimens (n = 14) from 7 MM patients before (i.e., baseline, 'day -4') and after (i.e., 'day 28') post-lymphodepleted BCMA CAR-T therapy. RESULTS: Our analysis revealed heterogeneity in gene expression profiles among MM cells, even those harboring the same cytogenetic abnormalities. The best overall responses (BORs) of patients over the 15-month follow-up are positively correlated with the abundance and targeted cytotoxic activity of CD8+ effector CAR-T cells on day 28 after CAR-T cell infusion. Additionally, favorable responses are associated with attenuated immunosuppression mediated by regulatory T cells (Tregs), enhanced CD8+ effector T cell cytotoxic activity, and elevated type 1 conventional dendritic cell (cDC1) antigen presentation ability. DC re-clustering inferred intramedullary-originated cDC3s with extramedullary migration. Cell-cell communication network analysis indicated BCMA CAR-T therapy mitigates BAFF/GALECTIN/MK pathway-mediated immunosuppression and activates MIF pathway-mediated anti-MM immunity. CONCLUSIONS: Our study sheds light on MM microenvironment dynamics after BCMA CAR-T therapy, offering clues for predicting treatment responsivity.

CDK7 inhibition induces apoptosis in acute myeloid leukemia cells and exerts synergistic antileukemic effects with azacitidine in vitro and in vivo.

THZ1, a CDK7 inhibitor, has potent antitumor effects in several cancers; however, its role in Acute myeloid leukemia (AML) is unclear. We explored the effects and potential mechanisms of THZ1, alone and in combination with azacitidine (AZA), in AML cells and xenograft models. THZ1 decreased cell viability, induced apoptosis in a dose and time-dependent manner, induced G0/G1 cell cycle arrest, decreased phosphorylated CDK1 and CDK2 expression, and inhibited RNA Pol II phosphorylation at multiple serine sites. The combination of AZA and THZ1 exhibited synergistic antileukemic effects in AML cell lines and primary cells with MCL1 and c-MYC downregulation. Moreover, the combination therapy significantly decreased tumor burden and prolonged animal survival in xenograft mice models. Our data demonstrate that CDK7 inhibition induces the apoptosis of AML cells and exerts a synergistic antileukemia effect with AZA in vitro and in vivo, which supports future exploration of this combination in clinical studies.

Human exposure to a mixture of endocrine disruptors and serum levels of thyroid hormones: A cross-sectional study.

Exposure to endocrine disruptors (EDCs) could disrupt thyroid hormone homeostasis. However, human epidemiological studies reported inconsistent observations, and scarce information on the effect of a mixture of chemicals. The aim of the present study was to examine the associations of multiple chemicals with thyroid hormones among adults from China. We measured serum levels of thyroid hormones and urinary levels of 11 EDCs, including six phthalate metabolites, bisphenol A (BPA), bisphenol F (BPF), bisphenol S (BPS), perchlorate, and thiocyanate among 177 healthy adults without occupational exposure. Associations of multiple urinary analytes with serum thyroid hormones were examined by performing general linear regression analysis and bayesian kernal machine regression analysis. These EDCs were detected in almost all samples. After adjusting for various covariates, we observed only BPF significantly associated with total thyroxin (TT4) (ß=-0.27, 95% confidence interval (CI) [-0.41, -0.14]), total triiodothyronine (TT3) (ß=-0.02 95% CI [-0.03, -0.01]), free T4 (fT4) (ß=-0.02, 95% CI [-0.03, -0.01]), and free T3 (fT3) (ß=-0.04, 95% CI [-0.07, -0.01]), and mono-(2-ethyl-5-oxohexyl) phthalate (MEOHP) and monoethyl phthalate (MEP) positively associated with TT4 (ß=0.24, 95% CI [0.01, 0.48]) and fT4 (ß=0.02, 95% CI [0.01, 0.04]), respectively. Moreover, we observed significant dose-response relationships between TT4 and the mixture of 11 EDCs, and BPF was the main contributor to the mixture effect, suggesting the priority of potential effect of BPF on disrupting thyroid function under a real scenario of human exposure to multiple EDCs. Our findings supported the hypothesis that human exposure to low levels of EDCs could alter thyroid hormones homeostasis among non-occupational healthy adults.

Acute myeloid leukemia with cup-like blasts and FLT3-ITD and NPM1 mutations mimics features of acute promyelocytic leukemia: a case of durable remission after sorafenib and low-dose cytarabine.

Some previous researches raised the possibility of a novel acute myeloid leukemia (AML) entity presenting cup-like cytomorphology with mutations of both FLT3 and NPM1 or one of them. However, the clinical implications of this subtype remain unknown. We describe a 63-year-old patient belonging to this distinct AML subtype, who presented similar features of acute promyelocytic leukemia (APL) including nuclear morphology, negative for CD34 and HLA-DR, and abnormal coagulation. He had no response to both arsenic trioxide and CAG regimen (cytarabine, aclarubicin, and G-CSF). Given that the patient carried the FLT3-ITD mutation, we switched to a pilot treatment of FLT3 inhibitor sorafenib combined with low-dose cytarabine (LDAC). To date, the patient achieved durable complete remission over 58 months. These findings suggest that AML with cup-like blasts and FLT3-ITD and NPM1 mutations mimic APL, and the prognosis of this subtype may be improved by sorafenib combined with LDAC.

Evaluation of efficacy and safety of DPP-4 inhibitors for Chinese elderly patients with type 2 diabetes mellitus.

BACKGROUND: The safety of hypoglycemic drugs should be paid more attention to in elderly patients with type 2 diabetes mellitus due to their concomitant diseases, physiological decline of liver and kidney function and cognitive decline. The aim of this study was to evaluate the efficacy and safety of DPP-4 inhibitors in elderly patients with type 2 diabetes mellitus. METHODS: From January 2010 to November 2018, 300 patients with type 2 diabetes mellitus who were over 60 years old were enrolled in the outpatient clinic of Geriatric Medical Center. Their medication records and follow-up medical records were used for retrospective analysis. The duration of treatment with DPP-4 inhibitors was more than 3 months. The changes of fasting blood glucose (GLU), glycosylated hemoglobin (HbA1C), body weight, body mass index (BMI) and liver and kidney function were compared before and after treatment. RESULTS: The average age of 300 patients (212 males and 88 females) was 73.7 ± 9.1 years old, BMI was 26.5 ± 2.8 kg/m2 and the duration of diabetes was 10.7 ± 8.2 years. The results of retrospective analysis showed that HbA1C decreased by 0.27% after treatment (P < 0.001). In the group of DPP-4 inhibitors used for less than 12 months, there was no difference in liver transaminase (ALT and AST) between before and after treatment, whereas in the group of DPP-4 inhibitors used formore than 12 months, liver transaminase decreased statistically compared with after treatment (P < 0.001). The incidence of fatty liver in elderly diabetic patients decreased after using DPP-4 inhibitors. There was no significant change in serum creatinine level and creatinine clearance rate in elderly patients with type 2 diabetes mellitus after treatment of DPP-4 inhibitor. In addition, the body weight and BMI of the patients decreased significantly (P < 0.001). No hypoglycemic reaction and gastrointestinal discomfort were found in the medical records. CONCLUSION: After DPP-4 inhibitors were used in elderly patients with type 2 diabetes mellitus, the elevated glycosylated hemoglobin could be controlled with improved safety of liver and kidney, and might have the effect of weight loss.

Augmented Rac1 Expression and Activity are Associated with Oxidative Stress and Decline of ß Cell Function in Obesity.

BACKGROUND: The aim of this study was to clarify the relationship among Rac1 expression and activation, oxidative stress and ß cell dysfunction in obesity. METHODS: In vivo, serum levels of glucose, insulin, oxidative stress markers and Rac1 expression were compared between ob/ob mice and C57BL/6J controls. Then, these variables were rechecked after the administration of the specific Rac1 inhibitor-NSC23766 in ob/ob mice. In vitro, NIT-1 ß cells were cultured in a hyperglycemic and/or hyperlipidemic state with or without NSC23766, and the differences of Rac1 expression and translocation, NADPH oxidase(Nox) enzyme activity, reactive oxygen species (ROS) and insulin mRNA were observed. RESULTS: ob/ob mice displayed abnormal glycometabolism, oxidative stress and excessive expression of Rac1 in the pancreas. NSC23766 injection inhibited the expression of Rac1 in the pancreas, along with amelioration of oxidative stress and glycometabolism in obese mice. Under hyperglycemic and/or hyperlipidemic conditions, Rac1 translocated to the cellular membrane, induced activation of the NADPH oxidase enzyme and oxidative stress, and simultaneously reduced the insulin mRNA expression in NIT-1 ß cells. Inhibiting Rac1 activity could alleviate oxidative stress and meliorate the decline of insulin mRNA in ß cells. CONCLUSIONS: Rac1 might contribute to oxidative stress systemically and locally in the pancreas in obesity. The excessive activation and expression of Rac1 in obesity were associated with ß cell dysfunction through ROS production.

[Monosomal karyotype among adult acute myeloid leukemia: clinical characteristic and prognostic analysis].

OBJECTIVE: To explore the clinical characteristics and prognostic value of monosomal karyotype (MK) patients in adult acute myeloid leukemia (AML). METHODS: We retrospectively studied 45 patients of MK⁺ in newly-diagnosed adult AML in our center from Oct 2000 to Dec 2012. Clinical characteristics, cytogenetic data and prognostic features were analyzed in the cohort of MK⁺ patients. RESULTS: MK was found in 45 patients (19.0%) of 237 newly-diagnosed adult AML with cytogenetic data available at diagnoses. Among these 45 cases, there were 28 male (62.2%) and 17 female (37.8%). Median age of MK⁺ patients at diagnose was 58(18-91) years old. The presence of -5(31.1%) and -7(17.8%) were the most common chromatid among MK⁺ AML patients. MK was much more prevalent among elderly patients. Among AML patients, the proportions of MK⁺ patients younger than 30, 30 to 59 and older than 60 years old groups were 11.5%, 17.7% and 22.4%, respectively. There was no difference between MK⁺ and MK⁻ patients in gender distribution (P=0.545). There was also no difference between MK⁺ and MK⁻ patients in the distribution of FAB castigation (P=0.239). Median survival of MK⁺ AML patients was 6.5 months. Cumulative 5-year overall survival (OS) of was 5.2%. Forty-three MK⁺ patients (43/45, 95.6%) also had a complex karyotype (CK). Two cases that did not meet the CK had not achieved complete remission (CR), and died within 6 months. There were 12 patients who were CK⁺ in 192 MK⁻ patients. The differences of OS and CR rates between MK⁺CK⁺ patients and MK⁻CK⁺ were statistically significant (P<0.05). CONCLUSION: The increased detection rate of MK with age was associated with lower CR and OS in AML patients.

[The cytogenetic characteristics of 178 acute myeloid leukemia patients].

OBJECTIVE: To explore the cytogenetic characteristics of acute myeloid leukemia (AML) patients. METHODS: The karyotype analysis was performed in 178 AML using the short-term culture of bone marrow cell and G-banding technique. RESULTS: Among the 178 patients, 171 had enough metaphases for analysis and 128 (74.9%) had clonal karyotypic abnormalities. Twenty-seven patients were secondary to myelodysplastic syndrome (MDS-AML), with 25 (92.6%) patients carrying clonal karyotypic abnormalities. Among the remaining 144 patients of de novo AML, 103 (71.5%) had clonal karyotypic abnormalities. The rate of abnormal clonal karyotype was higher in MDS-AML than that of de novo AML (P = 0.021). Among the 171 patients, 41 (24.0%) were in favorable risk group, 80(46.8%) in intermediate risk group and 50 (29.2%) in adverse risk group. t(15;17) was the most common chromosomal aberration. The majority intermediate risk chromosomal aberration was normal karyotype. The most common cytogenetic abnormality among adverse group was a complex karyotype. Adverse cytogenetic aberrations, such as -5/5q-, -7/7q-, frequently occurred in conjunction with one another as part of a complex karyotype. Totally 75 patients were 60 years or older, among them, 16.0% were in favorable risk group, 48.0% in intermediate risk group and 36.0% in adverse risk group. Among 96 younger patients, 30.2% were in favorable risk group, 45.8% in intermediate risk group and 24.0% in adverse risk group. The rate of favorable risk chromosomal aberration was lower in elder patients than in younger (P = 0.031). The rate of adverse risk chromosomal aberration and the rate of monosomal karyotype were higher in MDS-AML than in de novo AML patients (P < 0.001). CONCLUSIONS: The most common favorable, intermediate and adverse chromosomal aberrations were t(15;17), normal karyotype and complex karyotype respectively. The karyotype was poor in MDS-AML and elder AML patients.

[Expression of the P1B, P2A, P3AB and P3D protein of hepatitis A virus in prokaryotic cell and antigenicity analysis].

OBJECTIVE: To express P1B, P2A, P3AB and P3D cDNA gene fragments in prokaryotic system using thioredoxin fusion expression system; to investigate the antigenicity and application of recombinant protein. METHODS: By using PCR technique, P1B, P2A, P3AB and P3D gene fragments were cloned. Choosing M47 as the expressive vector, the recombinant plasmid P1B, P2A, P3AB and P3D was constructed and expressed in Escherichia coli after inducing by IPTG. By anion exchange and affinity chromatography, purified recombinant protein was obtained. By using Western Blot analysis and indirect ELISA to detect its antigenic activity. RESULTS: Four recombinant plasmids was proved to be constructed successfully by sequencing and the correct molecular weight of their expression products. Recombinant proteins were obtained in BL21 (DE3) and purified after Ni2+ affinity chromatography. Western Blot analysis and indirect ELISA showed that P2a had specific antigenicity. CONCLUSION: The P2a protein expressed in prokaryotic system was proved to have specific antigenicity. The indirect ELISA distinguished 24 positive sera from 24 negative sera. It is very likely that P2a can be an antigen to diagnose acute patients of hepatitis A and differentiate inactivated vaccine-induced immunity from an infection.