Target Selection for T-Cell Therapy in Epithelial Ovarian Cancer: Systematic Prioritization of Self-Antigens.

Adoptive T cell-receptor therapy (ACT) could represent a promising approach in the targeted treatment of epithelial ovarian cancer (EOC). However, the identification of suitable tumor-associated antigens (TAAs) as targets is challenging. We identified and prioritized TAAs for ACT and other immunotherapeutic interventions in EOC. A comprehensive list of pre-described TAAs was created and candidates were prioritized, using predefined weighted criteria. Highly ranked TAAs were immunohistochemically stained in a tissue microarray of 58 EOC samples to identify associations of TAA expression with grade, stage, response to platinum, and prognosis. Preselection based on expression data resulted in 38 TAAs, which were prioritized. Along with already published Cyclin A1, the TAAs KIF20A, CT45, and LY6K emerged as most promising targets, with high expression in EOC samples and several identified peptides in ligandome analysis. Expression of these TAAs showed prognostic relevance independent of molecular subtypes. By using a systematic vetting algorithm, we identified KIF20A, CT45, and LY6K to be promising candidates for immunotherapy in EOC. Results are supported by IHC and HLA-ligandome data. The described method might be helpful for the prioritization of TAAs in other tumor entities.

Metabolomic analysis of endometrial cancer by high-resolution magic angle spinning NMR spectroscopy.

PURPOSE: To analyze endometrial metabolite profiles between patients with endometrial cancer and controls. METHODS: Seventeen (17) women with endometrium cancer and 18 controls were enrolled in this study. 1H HR-MAS (High Resolution-Magic Angle Spinning) NMR (Nuclear Magnetic Resonance) spectroscopy data obtained from endometrial tissue samples of patients with endometrial cancer and control group were analyzed with bioinformatics methods. RESULTS: Principal component analysis (PCA) and the partial least squares discriminant analysis (PLS-DA) score plots obtained with the multivariate statistical analysis of pre-processed spectral data shows a separation between the samples from patients with endometrial cancer and controls. Analysis results suggest that the levels of lactate, glucose, o-phosphoethanolamine, choline, glycerophosphocholine, phosphocholine, leucine, isoleucine, valine, glutamate, glutamine, n-acetyltyrosine, methionine, taurine, alanine, aspartate and phenylalanine are increased in patients with endometrial cancer compared to the controls. CONCLUSION: The metabolomics signature of patients with endometrial cancer is different from that of benign endometrial tissue.

The effect of motivational video and nutrition on the non-stress test: a randomised controlled clinical trial.

This study aimed to determine the effect of motivational video stimulation and nutrition on the non-stress test (NST). The sample of the study consisted of 360 pregnant women in total, including two experimental groups and one control group (120 pregnant women in each group). After the pregnant women in the study were divided into three groups at a 1:1:1 ratio with the computer-assisted random sampling method, the draw method was used to determine the experimental and control groups. The pregnant women in the first experimental group were allowed to consume cake and juice 30 min before the NST procedure. The pregnant women in the second experimental group were also allowed to consume cake and juice, while they were also shown a video with the content of development and changes in the mother and the foetus during pregnancy, with relaxing music, for about 15-20 min. There was no intervention made in the control group. The data were collected using a Participant Information Form and an NST Findings Registry Form which were created by the researchers based on their review of the relevant literature. The data were analysed using arithmetic means, percentage distributions, ANOVA, Bonferroni and chi-squared tests. It was found that the mean number of foetal movements and the mean number of accelerations in the cake + juice and motivational video groups were higher than those in the control group (p < 0.001). Besides, the pregnant women in the experimental groups were found to have more reactive NST results. As a result, it was determined that the motivational video and cake + juice interventions improved the movement of the foetus, the number of accelerations and the ratio of reactivity in NST, but the two interventions did not have any superiority over each other. It is recommended to offer cake and juice to pregnant women before NST or have them watch a motivational video during NST, for having good NST results.Impact statementWhat is already known about the topic? The false-positive rates of the NST, which enables monitoring foetal movements and foetal heart rates, are high. These false-positive rates of the NST lead to many obstetric complications, in addition to increasing the rates of cesarean-section deliveries. Recent studies showed that there are methods and factors that increase foetal movements and shorten the application period of the NST. It was identified that food intake, music therapy, foetal vibroacoustic and halogen light stimulation before the NST increases foetal movements and shortens the application period of the NST.What this paper adds? In the study, it was found that watching motivational videos and consuming cake and juice increased the number of foetal movements, the number of accelerations and the rate of reactive NST results, but the two interventions did not have superiority over each other.What are the implications of these findings for clinical practice and/or further research? In order to increase reactive NST rates, it is recommended that pregnant women consume cake and fruit juice before the procedure and watch a motivational video during the NST procedure.

RETROSPECTIVE ANALYSIS OF BORDERLINE OVARIAN TUMORS: OUTCOMES AT A SINGLE CENTER.

We wanted to discuss our experiences in the approach to borderline ovarian tumors, which constitute a group different from epithelial ovarian tumors with respect to their biological structure in line with retrospective information gathered from our cases. A total of 25 patients operated on for the indication of adnexal masses diagnosed as borderline ovarian tumors based on frozen section results were included in our study. Patient age, tumor diameter, tumor markers and surgeries performed were discussed in the light of the literature. Statistical analyses were performed using the SPSS software. The patient mean age was 43.84±11.34 years. The mass was localized in the right (n=13), left (n=11) or both (n=1) adnexal regions. The mean tumor diameter was 12.9±5.84 cm. Histopathologic examination established the diagnosis of serous borderline (n=14 patients) and mucinous borderline (n=11) ovarian tumors. Although the results of our study are consistent with current literature data, a greater number of current studies should be performed on borderline ovarian tumors, which are defined as a class of tumors different from epithelial ovarian tumors.

The impact of thyroid-stimulating hormone levels in euthyroid women on intrauterine insemination outcome.

BACKGROUND: The aim of this study was to examine the effect of thyroid-stimulating hormone (TSH) levels on intrauterine insemination (IUI) outcomes among euthyroid women. METHODS: A retrospective cohort study was conducted. A total of 302 women who started their first IUI cycle in our fertility center were included in this study. The patients were categorized into two groups based on their preconception TSH values: 0.38-2.49 mIU/Land 2.50-4.99 mIU/L. The clinical pregnancy rate was the main outcome parameter. As secondary parameters, we evaluated the differences in spontaneous abortion rate, live-birth delivery rate, and perinatal outcomes according to the preconception TSH threshold (< 2.5 and < 5.00 mIU/L). RESULTS: There was no significant difference between the two groups with respect to clinical pregnancy, miscarriage, and live-birth rates with an odds ratio of 1.67 (95% CI: 0.79-3.53), 1.08 (95% CI: 0.09-13.1), and 1.79 (95% CI: 0.77-4.2), respectively. In addition, there were no significant differences in perinatal outcomes (gestation at delivery, birth weight, and neonatal intensive care unit-administration rate) between the two groups. CONCLUSIONS: Our findings indicate that among euthyroid patients, preconception TSH values in the high-normal range (between 2.5 and 4.9 mIU/L) do not have a negative effect on IUI outcomes. TRIAL REGISTRATION: This study is retrospectively registered by Ethical Review Board at Inonu University in 19th December 2017; Ethics approval no is 2017-27-20.

Treatment of unruptured cornual pregnancies by local injections of methotrexate or potassium chloride under transvaginal ultrasonographic guidance.

OBJECTIVE: To demonstrate the outcome of intralesional management and show the safety of local treatment of cornual pregnancy. METHODS: Eight patients were treated with local methotrexate or potassium chloride injection. All patients underwent transvaginal ultrasound examination and were diagnosed by the criteria defined by Timor-Tritsch. In the case of fetal heart beat observation, potassium chloride was injected; and in the case of no heart beat detection, methotrexate was used. A follicle aspiration needle was inserted directly into the gestational sac under transvaginal guidance. RESULTS: Although it has been considered to be a risk factor, none of the patients in our study had previous ectopic pregnancy, history of infertility / in vitro fertilization, or cornual pregnancy. One of the patients had a medical history of abortion. In four cases, methotrexate was injected, and three patients received potassium chloride as a local treatment. None of the patients had any complication in the peri- or postoperative period. CONCLUSION: Using a local approach, the treatment agent can reach the area of the cornual pregnancy in high concentrations. Based on this case series, a local approach seems to be an effective and fertility-sparing method for treating unruptured cornual pregnancies.

Combined metabolic activators improve metabolic functions in the animal models of neurodegenerative diseases.

BACKGROUND: Neurodegenerative diseases (NDDs), including Alzheimer's disease (AD) and Parkinson's disease (PD), are associated with metabolic abnormalities. Integrative analysis of human clinical data and animal studies have contributed to a better understanding of the molecular and cellular pathways involved in the progression of NDDs. Previously, we have reported that the combined metabolic activators (CMA), which include the precursors of nicotinamide adenine dinucleotide and glutathione can be utilized to alleviate metabolic disorders by activating mitochondrial metabolism. METHODS: We first analysed the brain transcriptomics data from AD patients and controls using a brain-specific genome-scale metabolic model (GEM). Then, we investigated the effect of CMA administration in animal models of AD and PD. We evaluated pathological and immunohistochemical findings of brain and liver tissues. Moreover, PD rats were tested for locomotor activity and apomorphine-induced rotation. FINDINGS: Analysis of transcriptomics data with GEM revealed that mitochondrial dysfunction is involved in the underlying molecular pathways of AD. In animal models of AD and PD, we showed significant damage in the high-fat diet groups' brain and liver tissues compared to the chow diet. The histological analyses revealed that hyperemia, degeneration and necrosis in neurons were improved by CMA administration in both AD and PD animal models. These findings were supported by immunohistochemical evidence of decreased immunoreactivity in neurons. In parallel to the improvement in the brain, we also observed dramatic metabolic improvement in the liver tissue. CMA administration also showed a beneficial effect on behavioural functions in PD rats. INTERPRETATION: Overall, we showed that CMA administration significantly improved behavioural scores in parallel with the neurohistological outcomes in the AD and PD animal models and is a promising treatment for improving the metabolic parameters and brain functions in NDDs.

Combined metabolic activators improve cognitive functions in Alzheimer's disease patients: a randomised, double-blinded, placebo-controlled phase-II trial.

BACKGROUND: Alzheimer's disease (AD) is associated with metabolic abnormalities linked to critical elements of neurodegeneration. We recently administered combined metabolic activators (CMA) to the AD rat model and observed that CMA improves the AD-associated histological parameters in the animals. CMA promotes mitochondrial fatty acid uptake from the cytosol, facilitates fatty acid oxidation in the mitochondria, and alleviates oxidative stress. METHODS: Here, we designed a randomised, double-blinded, placebo-controlled phase-II clinical trial and studied the effect of CMA administration on the global metabolism of AD patients. One-dose CMA included 12.35 g L-serine (61.75%), 1 g nicotinamide riboside (5%), 2.55 g N-acetyl-L-cysteine (12.75%), and 3.73 g L-carnitine tartrate (18.65%). AD patients received one dose of CMA or placebo daily during the first 28 days and twice daily between day 28 and day 84. The primary endpoint was the difference in the cognitive function and daily living activity scores between the placebo and the treatment arms. The secondary aim of this study was to evaluate the safety and tolerability of CMA. A comprehensive plasma metabolome and proteome analysis was also performed to evaluate the efficacy of the CMA in AD patients. RESULTS: We showed a significant decrease of AD Assessment Scale-cognitive subscale (ADAS-Cog) score on day 84 vs day 0 (P = 0.00001, 29% improvement) in the CMA group. Moreover, there was a significant decline (P = 0.0073) in ADAS-Cog scores (improvement of cognitive functions) in the CMA compared to the placebo group in patients with higher ADAS-Cog scores. Improved cognitive functions in AD patients were supported by the relevant alterations in the hippocampal volumes and cortical thickness based on imaging analysis. Moreover, the plasma levels of proteins and metabolites associated with NAD + and glutathione metabolism were significantly improved after CMA treatment. CONCLUSION: Our results indicate that treatment of AD patients with CMA can lead to enhanced cognitive functions and improved clinical parameters associated with phenomics, metabolomics, proteomics and imaging analysis. Trial registration ClinicalTrials.gov NCT04044131 Registered 17 July 2019, https://clinicaltrials.gov/ct2/show/NCT04044131.

Mortality after kidney transplantation: 10-year outcomes.

OBJECTIVES: In the past decade, advances in immunological therapy have increased the survival of kidney recipients and their grafts. However, it has not achieved the desired level of improvement. This study aims to reveal the mortality among kidney recipients. METHODS: Medical data of the patients, who had undergone kidney transplantation (KT) between November 2010 and December 2020, were retrospectively reviewed. Inclusion criteria were adult kidney recipients, who had died. Exclusion criteria were pediatric recipients, recipients of en bloc and dual KT, recipients with missing data, and recipients with a primary non-functioning graft. The recipients were grouped according to their donor type; Group 1 (from a living donor) and Group 2 (from a deceased donor). Subgroup analyses were done for mortality by time-period post-transplant and for infectious causes of mortality. RESULTS: Of 314 recipients, 35 (11.14%) died. Twenty-nine recipients were included in the study (Group 1: 17 and Group 2: 12). The most common cause of mortality was infection (58.6%), and the second was cardiovascular disease (CVD) (24.1%). Sepsis developed in 29.4% of infection-related deaths, while COVID-19 constituted 23.5% of infection-related deaths. CONCLUSION: Early diagnosis and treatment of infectious and CVD are important to improve survival in kidney recipients.

OBJETIVOS: En la última década, los avances en la terapia inmunológica han aumentado la supervivencia de los receptores de riñón y sus injertos. Sin embargo, no se pudo lograr el nivel de mejora deseado. Este estudio tiene como objetivo revelar la mortalidad entre los receptores de riñón. MATERIALES Y MÉTODOS: Se revisaron retrospectivamente los datos médicos de los pacientes, que se habían sometido a un trasplante de riñón entre Noviembre de 2010 y Diciembre de 2020. Los criterios de inclusión fueron los receptores de riñón adultos, que habían fallecido. Los criterios de exclusión fueron los receptores pediátricos, los receptores de trasplantes de riñón dual y en bloque, los receptores con datos faltantes y los receptores con un injerto primario no funcionante. Los receptores se agruparon según su tipo de donante; Grupo 1 (de un donante vivo) y Grupo 2 (de un donante fallecido). Se realizaron análisis de subgrupos para la mortalidad por período de tiempo posterior al trasplante y para las causas infecciosas de mortalidad. RESULTADOS: De 314 beneficiarios, 35 (11,14%) fallecieron. Se incluyeron 29 receptores en el estudio (Grupo 1:17; Grupo 2:12). La causa más común de mortalidad fue la infección (58,6%) y la segunda fue la enfermedad cardiovascular (24,1%). La sepsis se desarrolló en el 29,4% de las muertes relacionadas con la infección, mientras que el COVID-19 constituyó el 23,5% de las muertes relacionadas con la infección. CONCLUSIÓN: El diagnóstico y tratamiento tempranos de enfermedades infecciosas y cardiovasculares es importante para mejorar la supervivencia de los receptores de riñón.

Laparoscopic management of a translocated intrauterine device embedded in the gastric serosa.

Intrauterine devices (IUD) are the most common contraceptive methods all over the world. Besides many advantages, there are also some complications of this method. The most important complication of IUD is uterine perforation. The reported incidence of IUD perforation ranges from 0.2 to 9.6 per thousand insertions. In this article we would like to report a case that has an IUD migrated to the gastric serosa. A 41 years-old Turkish woman presented with abdominal pain. Plain X-ray of the abdomen demonstrated two IUDs, one in the upper quadrant of the abdomen, the other in the pelvic area and determined in the endometrium by TV-USG (transvaginal ultrasonography). Laparascopy was performed and the IUD, lying over the omentum of stomach was removed. This report demonstrates the first case in the literature that has an IUD embedded in the gastric serosa and also a second IUD in uterine cavity. The management was performed laparoscopically. No complication was observed during and after operation.

The relation between the presence of a giant primary ovarian leiomyoma and the occurrence of epilepsy as a paraneoplastic syndrome.

INTRODUCTION: Primary ovarian leiomyoma is a quite rare tumor and usually it is small and unilateral. Most cases are asymptomatic. This benign tumor is usually diagnosed incidentally on routine pelvic examination or in pathologic specimens at surgery or at autopsy. CASE PRESENTATION: In this case, a patient with her first epileptic seizure was admitted to our emergency department. Following the initial treatment in neurology clinics, patient claimed of an abdominal pain, so by abdominal ultrasonography, it was determined that the patient had a giant abdominal mass measured 30 x 28 x 15 cm in diameter which was clinically diagnosed as malign ovarian tumor. CONCLUSION: At laparotomy, a radical hysterectomy was performed. It was reported as ovarian leiomyoma on pathological evaluation. After 15 days of surgery, patient is discharged without any complications and also no more epileptic seizures.

Comparison of the demographic and clinical features of pregnant and non-pregnant patients undergoing appendectomy.

BACKGROUND: This retrospective study aims to compare the demographic and clinicopathological characteristics of the pregnant and non-pregnant patients who underwent appendectomy with a presumed diagnosis of acute appendicitis. METHODS: Between June 2009 and January 2019, 431 reproductive-aged of female patients underwent appendectomy with a presumed diagnosis of acute appendicitis. Patients were divided into two groups considering their pregnancy status: pregnant group (n=48) and non-pregnant group (n=383). Both groups were compared with respect to demographic, clinical and histopathological features. RESULTS: No statistically significant difference was found between pregnant and non-pregnant groups except total bilirubin level (p=0.019) and ultrasonographic findings (p=0.016). In the non-pregnant group, negative appendectomy and perforation rates were 26% and 10.5%, where these rates for the pregnant group were 20.8% and 4.2%. Sensitivity, specificity and accuracy rates of ultrasonography for the pregnant group were 50%, 100% and 58.5%, where these rates for the non-pregnant group were 67.3%, 57.9% and 65%. The pregnancy date was the first trimester in 52.1%, the second trimester in 29.2% and the third trimester in 16.7% of the pregnants. None of the term births (87.5%) resulted in neither a fetal nor a maternal complication. However, 12.5% of the preterm births resulted in neonatal mortality. CONCLUSION: Although not statistically significant, this study points out relatively lower rates of negative appendectomy and perforated acute appendicitis among pregnant patients, which is related to the overly attentive evaluation of pregnants admitted due to acute abdomen.

Neural precursor cell-expressed developmentally down-regulated 4-like: a new biomarker in the pathophysiology of endometrial cancer.

Objectives Endometrial cancer is the most frequent tumor of the female genital tract. Ubiquitin is a small protein (8.5 kDa) found in all eukaryotic cells, binds to substrate proteins via a three-phase enzymatic pathway referred to as ubiquitination and plays an important role in cellular stability. Neural precursor cell-expressed developmentally down-regulated 4-like (NEDD4L) functions in the last phase of this enzymatic process. In this study, we investigated NEDD4L protein expression in endometrial cancer. Methods The study participants were divided into patients with benign endometrial pathologies (Group 1, n = 23), patients with endometrial hyperplasia (Group 2, n = 21) and patients with endometrial cancer (Group 3, n = 20). NEDD4L expression was detected by immunohistochemical staining and H scores were calculated to standardize staining intensity. Statistical analysis was performed using SPSS 16.0. Results NEDD4L expression levels according to H scores were significantly lower in patients diagnosed with endometrial cancer compared with those with benign endometrial pathologies. Conclusion NEDD4L is involved in maintaining cell stability, and reduced NEDD4L expression as a result of gene mutation may disrupt this balance in favor of tumorigenesis.

Renal cell carcinoma diagnosed during pregnancy: a case report and literature review.

Diagnosing cancer during pregnancy is uncommon. Although pregnancies with concomitant malignancies have been reported, urological tumours are possibly the most rarely identified tumours during pregnancy. Renal cell carcinoma appears to be the most common urological malignancy during pregnancy. In this case report, we discuss successful management of a patient who was diagnosed with renal cell carcinoma during the antenatal period.

Assessment of relationships between novel inflammatory markers and presence and severity of preeclampsia: Epicardial fat thickness, pentraxin-3, and neutrophil-to-lymphocyte ratio.

OBJECTIVE: The aim of this study was to evaluate the relation of three new inflammatory markers with presence and severity of preeclampsia and to compare the predictive values of all markers for presence of this setting. METHODS: In this study, a total of 100 consecutive pregnants with a diagnosis of preeclampsia and 40 healthy pregnants between October 2014 and April 2015 were included. Epicardial fat tissue was calculated by two-dimensional transthoracic echocardiography, and pentraxin-3 and neutrophil-to-lymphocyte ratio were measured by using an enzyme-linked immunosorbent assay method and routine blood count analysis, respectively. RESULTS: Epicardial fat thickness (p < 0.001), pentraxin-3 (p < 0.001), and neutrophil-to-lymphocyte ratio (p < 0.001) were found to be significantly increased in the preeclampsia as compared to the healthy pregnants. Furthermore, epicardial fat thickness (p = 0.002), pentraxin-3 (p < 0.001), and neutrophil-to-lymphocyte ratio (p < 0.001) were significantly elevated in the severe preeclampsia compared to mild preeclampsia. In the multivariate analysis, epicardial fat thickness (p = 0.013), pentraxin-3 (p = 0.04), and neutrophil-to-lymphocyte ratio (p < 0.001) were found as significant independent predictors of presence of preeclampsia after adjusting for other risk factors. CONCLUSION: Epicardial fat thickness, neutrophil-to-lymphocyte ratio, and pentraxin-3 are important markers that provide an additional information beyond that provided by conventional methods in predicting presence and severity of preeclampsia.

The relationships between blood pressure, blood glucose, and bone mineral density in postmenopausal Turkish women.

BACKGROUND: Hypertension, diabetes mellitus, and osteoporosis are important comorbidities commonly seen in postmenopausal women. The aim of the present study was to investigate the relationships between blood pressure, blood glucose, and bone mineral density (BMD) in postmenopausal Turkish women. METHODS: In this cross-sectional study, 270 consecutive patients who were admitted to an outpatient clinic with vasomotor symptoms and/or at least 1 year of amenorrhea were included. The patients were categorized into three groups according to their blood pressure and metabolic status as follows: normotensive, hypertensive nondiabetics, and hypertensive diabetics. The T- and z-scores of the proximal femur and lumbar vertebrae were measured with the dual-energy X-ray absorptiometry method to assess the BMD of the study groups. RESULTS: Lumbar vertebral T-scores (P<0.001), lumbar vertebral z-scores (P<0.003), and proximal femoral T-scores (P<0.001) were demonstrated to be significantly lower in the hypertensive diabetic group compared to the hypertensive nondiabetic and normotensive groups. Systolic blood pressure was significantly inversely correlated with lumbar vertebral T-scores (r=-0.382; P=0.001), lumbar vertebral z-scores (r=-0.290; P=0.001), and proximal femoral T-scores (r=-0.340; P=0.001). Moreover, diastolic blood pressure was significantly inversely correlated with lumbar vertebral T-scores (r=-0.318; P=0.001), lumbar vertebral z-scores (r=-0.340; P=0.001), and proximal femoral T-scores (r=-0.304; P=0.001). Hypertension (odds ratio [OR]: 2.541, 95% confidence interval [CI]: 1.46-3.48, P=0.003), diabetes mellitus (OR: 2.136, 95% CI: 1.254-3.678, P=0.006), and age (OR: 1.069, 95% CI: 1.007-1.163, P=0.022) were found to be significant independent predictors of osteopenia in a multivariate analysis, after adjusting for other risk parameters. CONCLUSION: The present study is the first to evaluate the relationships between blood pressure, blood glucose, and BMD in postmenopausal Turkish women. Moreover, both hypertension and diabetes were demonstrated as significant independent predictors of osteopenia in postmenopausal Turkish women. Clinicians should be aware of the high risk of developing osteopenia in diabetic hypertensive postmenopausal women.

MicroRNA profiling reveals aberrant microRNA expression in adult ETP-ALL and functional studies implicate a role for miR-222 in acute leukemia.

Early T-cell precursor acute lymphoblastic leukemia (ETP-ALL) has been identified as high-risk subgroup in acute T-cell lymphoblastic leukemia (T-ALL). To investigate the immature and myeloid nature of ETP-ALL we examined global microRNA (miRNA) expression in adult ETP-ALL. miRNA profiling of ETP-ALL (n=8), non-ETP T-ALL (n=6), and healthy controls was performed and results were validated in independent cohorts of 66 ETP-ALL and 111 non-ETP T-ALL using real-time RT-PCR. Furthermore, in vitro studies were performed on deregulated miRNAs in acute leukemia. We identified miR-221 and miR-222 as the most upregulated and six miRNAs (miR-151-3p, miR-19a, miR-20b, miR-342-3p, miR-363, and miR-576-3p) as downregulated in ETP-ALL compared to non-ETP T-ALL. In the validation cohorts, miR-221 and miR-222 were significantly upregulated in ETP-ALL, and miR-363 and miR-19a were downregulated in ETP-ALL. ETS1, downregulated in ETP-ALL, was identified as direct target of miR-222. In our in vitro studies miR-222 significantly inhibited proliferation, and caused cell cycle arrest and apoptosis in leukemic cells. In conclusion, our study revealed aberrant miRNA expression in ETP-ALL, with miR-221 and miR-222 as the most overexpressed miRNAs and implied a functional role for miR-222 in leukemic cells. Importantly, miR-222 may impact leukemogenesis by altering expression of the proto-oncogene ETS1 in acute leukemia.

FLT3 mutations in early T-cell precursor ALL characterize a stem cell like leukemia and imply the clinical use of tyrosine kinase inhibitors.

Early T-cell precursor acute lymphoblastic leukemia (ETP-ALL) has been identified as high-risk subgroup of acute T-lymphoblastic leukemia (T-ALL) with a high rate of FLT3-mutations in adults. To unravel the underlying pathomechanisms and the clinical course we assessed molecular alterations and clinical characteristics in a large cohort of ETP-ALL (n = 68) in comparison to non-ETP T-ALL adult patients. Interestingly, we found a high rate of FLT3-mutations in ETP-ALL samples (n = 24, 35%). Furthermore, FLT3 mutated ETP-ALL was characterized by a specific immunophenotype (CD2+/CD5-/CD13+/CD33-), a distinct gene expression pattern (aberrant expression of IGFBP7, WT1, GATA3) and mutational status (absence of NOTCH1 mutations and a low frequency, 21%, of clonal TCR rearrangements). The observed low GATA3 expression and high WT1 expression in combination with lack of NOTCH1 mutations and a low rate of TCR rearrangements point to a leukemic transformation at the pluripotent prothymocyte stage in FLT3 mutated ETP-ALL. The clinical outcome in ETP-ALL patients was poor, but encouraging in those patients with allogeneic stem cell transplantation (3-year OS: 74%). To further explore the efficacy of targeted therapies, we demonstrate that T-ALL cell lines transfected with FLT3 expression constructs were particularly sensitive to tyrosine kinase inhibitors. In conclusion, FLT3 mutated ETP-ALL defines a molecular distinct stem cell like leukemic subtype. These data warrant clinical studies with the implementation of FLT3 inhibitors in addition to early allogeneic stem cell transplantation for this high risk subgroup.