Primary hyperoxaluria type 1 (PH1) is a rare autosomal recessive disorder of the liver metabolism due to functional deficiency of the peroxisomal enzyme alanine:glyoxylate aminotransferase (AGT). AGT ...deficiency results in overproduction of oxalate which complexes with calcium to form insoluble calcium-oxalate salts in urinary tracts, ultimately leading to end-stage renal disease. Currently, the only curative treatment for PH1 is combined liver-kidney transplantation, which is limited by donor organ shortage and lifelong requirement for immunosuppression. Transplantation of genetically modified autologous hepatocytes is an attractive therapeutic option for PH1. However, the use of fresh primary hepatocytes suffers from limitations such as organ availability, insufficient cell proliferation, loss of function, and the risk of immune rejection. We developed patient-specific induced pluripotent stem cells (PH1-iPSCs) free of reprogramming factors as a source of renewable and genetically defined autologous PH1-hepatocytes. We then investigated additive gene therapy using a lentiviral vector encoding wild-type AGT under the control of the liver-specific transthyretin promoter. Genetically modified PH1-iPSCs successfully provided hepatocyte-like cells (HLCs) that exhibited significant AGT expression at both RNA and protein levels after liver-specific differentiation process. These results pave the way for cell-based therapy of PH1 by transplantation of genetically modified autologous HLCs derived from patient-specific iPSCs.
•Primary hyperoxaluria (PH1) is an inherited disease amenable to liver-targeted ex-vivo gene therapy.•Sendaï vectors allow the generation of viral-free PH1-induced pluripotent stem cells from dermal fibroblasts.•PH1-induced pluripotent stem cells differentiate into functional hepatocytes-like cells.•Hepatic-specific lentiviral vector efficiently rescue AGT expression in hepatocyte like-cells.
Chromosomal rearrangements of the human KMT2A/MLL gene are associated with de novo as well as therapy-induced infant, pediatric, and adult acute leukemias. Here, we present the data obtained from ...3401 acute leukemia patients that have been analyzed between 2003 and 2022. Genomic breakpoints within the KMT2A gene and the involved translocation partner genes (TPGs) and KMT2A-partial tandem duplications (PTDs) were determined. Including the published data from the literature, a total of 107 in-frame KMT2A gene fusions have been identified so far. Further 16 rearrangements were out-of-frame fusions, 18 patients had no partner gene fused to 5'-KMT2A, two patients had a 5'-KMT2A deletion, and one ETV6::RUNX1 patient had an KMT2A insertion at the breakpoint. The seven most frequent TPGs and PTDs account for more than 90% of all recombinations of the KMT2A, 37 occur recurrently and 63 were identified so far only once. This study provides a comprehensive analysis of the KMT2A recombinome in acute leukemia patients. Besides the scientific gain of information, genomic breakpoint sequences of these patients were used to monitor minimal residual disease (MRD). Thus, this work may be directly translated from the bench to the bedside of patients and meet the clinical needs to improve patient survival.
HLA immunization is a common complication of transfusion therapy in 30% to 60% of oncohematologic patients. Evidence shows that leukocytes present in cellular blood products are the main component ...involved in the occurrence of HLA immunization, and several studies showed that leukocyte-poor blood products are less able to induce it. However, leukocyte-poor platelet concentrates obtained by conventional techniques, ie, centrifugation, frequently have a high level of remaining leukocytes. Cotton wool filter Imugard IG 500 can be used to obtain leukocyte-poor cellular blood products. The technique is easy to perform, even in an emergency, and can be used with either packed RBCs or platelet concentrates. Means of 97%, 92%, and 76% elimination of leukocytes are obtained for packed RBCs, pooled standard platelet concentrates, and single-donor platelet concentrates, respectively. Patients were randomized to receive either standard (con-trol group) or filtered (leukocyte-poor group) blood products. Of 112 randomized patients, 69 were evaluable, 35 in the control group and 34 in the leukocyte-poor group. Both groups are comparable according to age, diagnosis, sex ratio, previous transfusions, and pregnancies. There is a significant difference in regard to the HLA immunization rate (31.4% in the control v 11.7% in the leukocyte-poor group, P < .05) and frequency of refractoriness to platelet transfusions (46.6% v 11.7%, P < .05). We conclude that this filtration technique can be an efficient means to reduce the HLA immunization rate in polytransfused oncohemato-logic patients.1988 by Grune & Stratton, Inc. 0006-4971/88/7203-0023$3.00/0
BACKGROUND: Despite blood donor screening, there are still cases of transfusion‐associated hepatitis. From 1988 to 1992, a prospective study was conducted on the incidence of non‐A, non‐B ...posttransfusion hepatitis (PTH). STUDY DESIGN: The present investigation was designed to determine if transfusion recipients with PTH who are negative for hepatitis C virus (HCV) were positive for hepatitis G virus (HGV). Patients admitted for surgery who had normal liver tests and no transfusions during the previous 6 months were enrolled. Alanine amino transferase levels were determined monthly for 6 months after surgery and for 1 year in the case of PTH (defined as alanine aminotranferase twice the upper limit of normal in two consecutive assays). HGV RNA and E2 antibodies were tested for in samples from transfusion recipients with or without PTH and from nontransfused patients. RESULTS: Of the 308 blood recipients who were enrolled in the study, 21 (6.8%) had PTH. HGV RNA was detected at the onset of hepatitis in 3 patients with PTH (14%), 2 of whom were also anti‐HCV and HCV RNA positive. One patient developed E2 antibodies without detectable HGV RNA. Three (10.7%) of 28 recipients of an allogeneic transfusion without PTH developed HGV infection. HGV RNA was also found in two nontransfused patients, which suggests nosocomial transmission of HGV. CONCLUSION: Some cases of PTH are associated with HGV; most cases of postoperative HGV infection are not associated with liver abnormalities; and most PTH cases are not associated with known hepatotropic viruses.
Abstract
Background: Panitumumab is an antibody targeting the epidermal growth factor receptor (EGFR) for which an important role has been suggested in TNBC. Consequently, we evaluated a combination ...of the standard chemotherapy (FEC 100 followed by docetaxel) with panitumumab as neoadjuvant therapy of operable TNBC. Complete pathologic response (pCR) was the primary endpoint, with clinical response, toxicity, and outcome as secondary endpoints. An investigation of biomarkers possibly predictive of pCR accompanied this trial. Here we focus on tumor recurrence, after a median follow up of 33 months 25-40 as on April, 1, 2013.
Methods: Sixty patients (pts) with stage II-IIIA TNBC were prospectively included. Systemic neoadjuvant treatment (ST) consisted of the anti-EGFR antibody panitumumab combined with FEC 100, followed by 4 cycles of docetaxel. All pts underwent surgery after ST completion. Patient characteristics: median tumor size: 40 mm 20-120; SBR grade III: 71.7%; pCR rate: 55.3% and 46.8% (the Sataloff and the Chevallier classifications, respectively). Paraffin-embedded and frozen tumor samples were collected before and after ST for biomarker analysis. EGFR, IGF-1R, MET, cytokeratins 5/6 and 8/18, PTEN, P-cadherin, ALDH1, Ki-67, p53, tumoral FOXP3 expression and the number of FOXP3+ or CD8+ tumor-infiltrating lymphocytes (TIL) were evaluated by immunohistochemistry.
Results :.We have observed 9 recurrences: 1 local and 8 distant recidives, including 1 both local and distant.
The distant recidives (metastases) were as follows: brain (4 pts); brain and lungs (1 pt); lungs only (1 pt), pleura (1 pt); liver (1 pt). 6 out of the 8 metastatic pts died and all were non-pCR post-ST. The 2 alive pts had brain metastases, but reached a pCR after the ST.
Among the 9 relapsed pts 6 were 55 years old or less at the diagnosis. Seven out of those 9 pts had tumors with the clinical size equal or higher than 4 cm.
As previously reported (SABCS 2012, abstract 1081), the pCR-predictive biomarkers in this study were high CD8+ TIL count (p = 3.4.10−6) and high ratio between the CD8+ and FOXP3+ TIL counts (CD8+/FOXP3+ > 1.23, p = 8.5.10−5). With this in mind, we have evaluated whether those parameters, assessed before or after the ST, could predict the recurrences. No difference was found in the preoperative CD8+ and the FOXP3+ TIL counts, as well as in the CD8+/FOXP3+ ratio, between the patiens who have recurred and the others.
Conclusion : As it has been reported in previous studies, in our cohort of TNBC pts, the relapses occurred early after the administration of the last systemic treatment. The patients who relapsed died rapidly and most of them have not reached pCR after the ST. In addition, half of the metastatic pts got brain deposits. This implies that research on the resistance factors in TNBC should focus on those important for seeding of the “sanctuaries”, like brain. This research is ongoing in our group.
Citation Information: Cancer Res 2013;73(24 Suppl): Abstract nr P3-14-19.
Abstract
Background: TNBC is a heterogenous group of tumors for some of which the Epithelial Growth Factor Receptor pathway (EGFR) may play an important role. We evaluated the efficacy and toxicity ...of an anti-EGFR antibody (cetuximab) combined with docetaxel, which were given to the TNBC patients (pts) in the neoadjuvant setting. A biomarker analysis accompanied this trial, aiming to identify predictors of response.
Methods: 35 patients with stage II-IIIA TN breast disease were prospectively included in this multicentre pilot study. Systemic therapy (ST) consisted of 6 cycles of docetaxel (100 mg/m2) each 3 weeks, in combination with weekly cetuximab (first dose 400mg/m2 then 250 mg/m2/week) for 6 cycles. All patients underwent surgery at completion of ST.
Patient characteristics : mean age 48 28-67; TNM: T1: 3%, T2: 73%, T3: 24%; N0: 61%, N1-N2: 39%; mean tumor size 40 mm 15-100; SBR: grade III: 73%, grade II: 27%. The median number of cycles was for docetaxel 6 1-6 and for cetuximab 15 1-18. Pathological complete response (pCR) rate was 24% according to the Chevallier and Sataloff classifications; 28% if we consider breast pCR only. Overall clinical response rate was 57% (22% CR). Conservative surgery was performed in 75% of cases. The main side effect was skin toxicity: grade II: 39%, grade III: 36%, grade IV: 3%. Other side effects were: neutropenia grade IV: 12.7%, febrile neutropenia: 1.3%, hand-foot syndrome grade III: 3%, grade II: 3%, ungueal toxicity grade III: 3%, grade II: 33%.
Paraffin-embedded and frozen samples were systematically collected before and after ST for biomarker studies. Germinal BRCA1 mutations and EGFR, KRAS, BRAF and PI3KCA somatic mutations were analyzed by NGS. EGFR, MET, cytokeratins 5/6 and 8/18, PTEN, P-cadherin, ALDH1, Ki67, p53 and the number of FOXP3+ or CD8+ tumor-infiltrating lymphocytes (TIL) were evaluated by immunohistochemistry.
Results: The biomarker analysis was interpretable on the samples from 21 pts (3 pCR and 18 non-pCR).
We applied the ROC curve to identify the best cut-off value for Ki67, EGFR, MET, cytokeratin 5/6 and 8/18, p53, ALDH1, PTEN, P-cadherin and the FOXP3+ or CD8+ TIL counts. None of these biomarkers was predictive of pCR except for the CD8+/FOXP3+ TIL count ratio. pCR rate was higher in the pts with the ratio equal or higher than 2.75 than in the others (43% versus 0%, p = 0.047).
BRCA1 mutations were detected in 16% of pts. PI3K and EGFR somatic mutations were observed in 1 and 3 patients, respectively. The presence of the mutations was not predictive of pCR.
Conclusion: Similarly to the previously reported trial by our group (SABCS 2012, abstract 1081), the immune component of the tumor microenvironment plays a very important role in the TNBC response to cytotoxic therapy.
Citation Information: Cancer Res 2013;73(24 Suppl): Abstract nr P1-08-34.
Currently, no single diagnostic modality allows the distinction between early progression (EP) and pseudo-progression (Psp) in glioblastoma patients. Herein we aimed to identify the characteristics ...associated with EP and Psp, and to analyze their diagnostic value alone and in combination.
We reviewed the clinical, conventional magnetic resonance imaging (MRI), and molecular characteristics (MGMT promoter methylation, IDH mutation, and EGFR amplification) of glioblastoma patients who presented an EP (n=59) or a Psp (n=24) within six months after temozolomide radiochemotherapy. We analyzed relative cerebral blood volume (rCBV) and relative vessel permeability on K2 maps (rK2) in a subset of 33 patients using dynamic-susceptibility-contrast MRI.
In univariate analysis, EP was associated with neurological deterioration, higher doses of dexamethasone, appearance of a new enhanced lesion, subependymal enhancement, higher rCBV and rK2 values. Psp occurred earlier after radiotherapy completion and was associated with IDH1 R132H mutation, and MGMT methylation. In multivariate analysis, rCBV, rK2, and MGMT methylation status were independently associated with EP and Psp. All patients with a methylated MGMT promoter and a low rCBV (<1.75) were classified as Psp while all patients with an unmethylated MGMT promoter and a high rCBV (≥1.75) were classified as EP. Among patients with discordant MGMT methylation and rCBV characteristics, higher rK2 values tended to be associated with EP.
Combined analysis of MGMT methylation, rCBV and vessel permeability on K2 maps seems helpful to distinguish EP from Psp. A prospective study is warranted to confirm these results.
Abstract
Study question
Is ovarian stimulation (OS) with r-hFSH:r-hLH more effective than OS with r-hFSH alone in women aged 35-42 with normal ovarian reserve treated with ART?
Summary answer
...Compared to r-hFSH alone, OS with r-hFSH:r-hLH for ART treatment was associated with a significant increase in clinical pregnancy and live birth per initiated cycle.
What is known already
Meta-analyses and secondary analyses of clinical data provide evidence that OS during ART treatment with r-hFSH:r-hLH is associated with improved reproductive outcomes when compared to r-hFSH alone in various patient groups with: a) advanced maternal age , b) low ovarian reserve or , c) hypo response to OS. Other subgroups of patients might also benefit of the use of r-hFSH:r-hLH during OS for ART treatment. We aimed to determine if, in a real-world setting, OS with r-hFSH:r-hLH is associated with improved clinical benefit when compared to r-hFSH alone in patients aged 35-42 years with normal ovarian reserve.
Study design, size, duration
A non-interventional study including women aged 35-42 years with normal ovarian reserve biomarkers (menstrual cycle day 2 or 3 FSH<12IU/L or 9<AFC<20 with a 2<diameter<10 mm, or AMH ≥1.2ng/ml) undergoing OS for ART treatment (fresh transfer only; GnRH antagonist protocol) in 12 French centers between 1/1/2008 and 31/12/2016, with a follow-up period up to 31/12/2017. The analysis included two cohorts: a) 4,323 OS cycles with r-hFSH alone, and b) 1,070 OS cycles with r-hFSH:r-hLH.
Participants/materials, setting, methods
Anonymized data were extracted from the Retrospective Multicenter Statistical database, including 12 French centers. A generalized linear model was used to address lack of randomization. The fixed effect part included age, an ovarian reserve estimator and stimulation; the random effect included age, AMH, AFC, FSH, cause of infertility and number of attempts, center and patient effect. Clusters (matched subsets) were built based on the propensity score by using a two-stage algorithm.
Main results and the role of chance
We included in our analysis 4,472 women receiving ART treatment after OS with r-hFSH (mean age: 37.8 years; 4,323 cycles) or with r-hFSH:r-hLH (mean age: 38.1 years; 1,070 cycles). In the r-hFSH cohort the mean AMH level was 2.81 ng/ml and mean AFC was 12.0, in the r-hFSH:r-hLH cohort they were 2.14 ng/ml and 10.0, respectively. The mean total r-hFSH dose were 2,275 IU (r-hFSH cohort) and 3,002 IU (r-hFSH:r-hLH cohort). Mean ± SD number of oocytes were 8.1 ± 5.7 (r-hFSH cohort) and 6.1 ± 4.3 (r-hFSH:r-hLH cohort); mean ± SD number of embryos were 4.0 ± 3.4 (r-hFSH cohort) and 3.3 ± 3.2 (r-hFSH:r-hLH cohort). The unadjusted ongoing pregnancy rate reached 17.1% in the r-hFSH cohort and 20.4% in the r-hFSH:r-hLH cohort, whilst the unadjusted live birth rate reached 17.0% and 20.2%, respectively. Unadjusted pregnancy failure rate was 6.6% and 3.7%, respectively.
After adjusting for confounders, when compared to OS with r-hFSH, OS with rhFSH:rhLH was associated with a higher clinical pregnancy rate (Risk ratio,RR 1.232; 95% CI 1.058 to 1.436; p-value = 0.007) and a higher live birth rate (RR 1.452; 95% CI 1.227-1.1719, p-value <0.001) per initiated cycle.
Limitations, reasons for caution
Since this was a real-world data study, no random assignment of stimulation was performed, and some residual confounding is possible due to unmeasured confounders. Live births resulting from further transfers of frozen embryos were not included in the analysis.
Wider implications of the findings
This study demonstrated that OS with r-hFSH:r-hLH improves live birth rate in women > 35 years old with a normal ovarian reserve, compared to r-hFSH only. Sound RWE studies can be combined with RCTs to guide clinical and payer’s decision making, paving the way to precision medicine.
Trial registration number
Not applicable
Clustered cases of Pseudomonas aeruginosa infection in immunocompromised patients' wards require rapid characterization of a potential epidemic to guide investigations and identify the potential ...source of contamination.
To design and evaluate a rapid and simple typing method for P. aeruginosa in comparison to whole genome sequencing (WGS).
A simplified polymerase chain reaction based on multiple-locus variable-number of tandem repeats analysis (MLVA) was designed and used to investigate cases of P. aeruginosa infection and colonization in a paediatric haematology department. The method was compared to WGS by using the Illumina method.
On the 17 isolates recovered from 15 children (eight from blood cultures, three from urinary tract infections, one from sputum and five stool isolates), MLVA distinguished 10 different profiles, and seven isolates from six children shared the same profile. Analysis by WGS revealed that these seven isolates belonged to sequence type ST111 and serotype O12, allowing at least three different genotypes to be distinguished among them. Five environmental strains had three MLVA profiles; one was shared with a clinical isolate but WGS excluded any relationship.
The simplified and inexpensive MLVA method enabled the exclusion, in less than 5 h, of most of the unrelated isolates and thus to focus investigations on a small number of cases, whereas WGS, taking several days of work, drew definitive conclusions concerning the outbreak and the genetic relationships of the ST111 isolates circulating in the department. We conclude that sequential use of both methods is the optimal strategy to investigate clustered cases of P. aeruginosa infections.
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