Molecular Criteria for Defining the Naive Human Pluripotent State.

Recent studies have aimed to convert cultured human pluripotent cells to a naive state, but it remains unclear to what extent the resulting cells recapitulate in vivo naive pluripotency. Here we propose a set of molecular criteria for evaluating the naive human pluripotent state by comparing it to the human embryo. We show that transcription of transposable elements provides a sensitive measure of the concordance between pluripotent stem cells and early human development. We also show that induction of the naive state is accompanied by genome-wide DNA hypomethylation, which is reversible except at imprinted genes, and that the X chromosome status resembles that of the human preimplantation embryo. However, we did not see efficient incorporation of naive human cells into mouse embryos. Overall, the different naive conditions we tested showed varied relationships to human embryonic states based on molecular criteria, providing a backdrop for future analysis of naive human pluripotency.This study was supported by grants from the Simons Foundation (SFLIFE #286977 to R.J) and in part by the NIH (RO1-CA084198) to R.J., from the Swiss National Science Foundation and the European Research Council (KRABnKAP, No. 268721) to D.T. The work in J.R.E’s laboratory was supported by the Howard Hughes Medical Institute and Gordon and Betty Moore Foundation (GBMF3034) and the Mary K. Chapman Foundation. J.R.E is an Investigator of the Howard Hughes Medical Institute. T.W.T. is supported by a Sir Henry Wellcome Postdoctoral Fellowship (098889/Z/12/Z), J.P. by a Foundation Bettencourt Award and by the Association pour la Recherche sur le Cancer (ARC), M.I. by a postdoctoral training grant from the Fonds de la Recherche en Santé du Québec. R.J. is co-founder of Fate Therapeutics and an adviser to Stemgent.This is the final version of the article. It first appeared from Cell Press via http://www.cell.com/cell-stem-cell/abstract/S1934-5909(16)30161-

Safety and efficacy of a single intraarticular injection of a novel enhanced protein solution (JTA-004) compared to hylan G-F 20 in symptomatic knee osteoarthritis: a randomized, double-blind, controlled phase II/ III study

peer reviewedBackground: New minimally invasive treatments are vital to delay joint replacement surgery in patients with knee osteoarthritis. This study was designed to select the most effective among three formulations of an enhanced protein solution containing clonidine, hyaluronic acid, and human plasma (JTA-004), and compare the safety and efficacy of intra-articular administration of the selected formulation with a reference treatment (hyaluronic acid) in symptomatic knee osteoarthritis patients. Methods: In this two-stage, double-blind, phase II/III study conducted in 12 Belgian centers, 50–79 year-old patients with primary knee osteoarthritis were randomized (1:1:1:1) to receive one dose of one of three JTA 004 formulations (differing in clonidine concentration [50 or 100 μg/ml] and volume [2 or 4 ml]) or the reference treatment (hylan G-F 20). Patients were evaluated using Western Ontario McMaster Universities (WOMAC®) Scores and the Short-Form health survey up to 6 months post-injection (Month 6). Drug consumption and safety were evaluated. Results: Among 164 treated patients, 147 completed the study. The JTA-004 formulation containing 200 μg clonidine and 20 mg hyaluronic acid in 2 ml (JTA-200/2) was selected based on interim results at Month 6. The difference in adjusted mean change in WOMAC Pain Subscale Score from baseline (JTA-200/2 minus reference group) at Month 6 was − 9.49 mm; statistical superiority of JTA-200/2 over the reference was not demonstrated. No statistically significant differences in adjusted mean changes from baseline between JTA-200/2 and reference groups were observed for Pain, Physical Function and Stiffness Subscales WOMAC Scores, Total WOMAC Score, and Well being Score at any timepoint, although JTA-200/2 induced larger improvements in WOMAC Scores than the reference. Statistically significantly larger improvements in WOMAC Pain Subscale Scores for JTA-004 versus the reference were observed in post-hoc analyses on pooled data from all JTA-004 formulations at Month 6 (p = 0.030) and Month 3 (p = 0.014). All JTA-004 formulations had clinically acceptable safety profiles. Conclusions: This study provided preliminary evidence of the safety of intra-articular injection of JTA 004 in knee osteoarthritis patients. Phase III randomized controlled trials with larger sample sizes are needed to evaluate the efficacy of JTA-004 in knee osteoarthritis

Hominoid-Specific Transposable Elements and KZFPs Facilitate Human Embryonic Genome Activation and Control Transcription in Naive Human ESCs

Expansion of transposable elements (TEs) coincides with evolutionary shifts in gene expression. TEs frequently harbor binding sites for transcriptional regulators, thus enabling coordinated genome-wide activation of species- and context-specific gene expression programs, but such regulation must be balanced against their genotoxic potential. Here, we show that Krüppel-associated box (KRAB)-containing zinc finger proteins (KZFPs) control the timely and pleiotropic activation of TE-derived transcriptional cis regulators during early embryogenesis. Evolutionarily recent SVA, HERVK, and HERVH TE subgroups contribute significantly to chromatin opening during human embryonic genome activation and are KLF-stimulated enhancers in naive human embryonic stem cells (hESCs). KZFPs of corresponding evolutionary ages are simultaneously induced and repress the transcriptional activity of these TEs. Finally, the same KZFP-controlled TE-based enhancers later serve as developmental and tissue-specific enhancers. Thus, by controlling the transcriptional impact of TEs during embryogenesis, KZFPs facilitate their genome-wide incorporation into transcriptional networks, thereby contributing to human genome regulation. Transposable elements (TEs) are key to the evolutionary turnover of regulatory sequences but potentially toxic to the host. Trono and colleagues demonstrate that KRAB zinc-finger proteins tame the activity of TEs during human early embryogenesis, thus allowing for their genome-wide incorporation into species-specific transcriptional networks.National Institutes of Health (Grant R37HD045022)National Institutes of Health (Grant R01-NS088538

Rapid, postmortem 9.4 T MRI of spinal cord injury: correlation with histology and survival times.

High field magnetic resonance imaging (MRI) has been increasingly used to assess experimental spinal cord injury (SCI). In the present investigation, after partial spinal cord injury and excision of the whole spine, pathological changes of the spinal cord were studied in spinal cord-spine blocks, from the acute to the chronic state (24 h to 5 months). Using proton density (PD) weighted imaging parameters at a magnetic field strength of 9.4 tesla (T), acquisition times ranging from <1 to 10 h per specimen were used. High in-plane pixel resolution (68 and 38 microm, respectively) was obtained, as well as high signal-to-noise ratio (SNR), which is important for optimal contrast settings. The quality of the resulting MR images was demonstrated by comparison with histology. The cord and the lesion were shown in their anatomical surroundings, detecting cord swelling in the acute phase (24 h to 1 week) and cord atrophy at the chronic stage. Haemorrhage was detected as hypo-intense signal. Oedema, necrosis and scarring were hyper-intense but could not be distinguished. Histology confirmed that the anatomical delimitation of the lesion extent by MRI was precise, both with high and moderate resolution. The present investigation thus demonstrates the precision of spinal cord MRI at different survival delays after compressive partial SCI and establishes efficient imaging parameters for postmortem PD MRI

Safety and efficacy of a single intra-articular injection of a novel enhanced protein solution (JTA-004) compared to hylan G-F 20 in symptomatic knee osteoarthritis: a randomized, double-blind, controlled phase II/III study

Background: New minimally invasive treatments are vital to delay joint replacement surgery in patients with knee osteoarthritis. This study was designed to select the most effective among three formulations of an enhanced protein solution containing clonidine, hyaluronic acid, and human plasma (JTA-004), and compare the safety and efficacy of intra-articular administration of the selected formulation with a reference treatment (hyaluronic acid) in symptomatic knee osteoarthritis patients. Methods: In this two-stage, double-blind, phase II/III study conducted in 12 Belgian centers, 50–79-year-old patients with primary knee osteoarthritis were randomized (1:1:1:1) to receive one dose of one of three JTA-004 formulations (differing in clonidine concentration [50 or 100 μg/ml] and volume [2 or 4 ml]) or the reference treatment (hylan G-F 20). Patients were evaluated using Western Ontario McMaster Universities (WOMAC®) Scores and the Short-Form health survey up to 6 months post-injection (Month 6). Drug consumption and safety were evaluated. Results: Among 164 treated patients, 147 completed the study. The JTA-004 formulation containing 200 μg clonidine and 20 mg hyaluronic acid in 2 ml (JTA-200/2) was selected based on interim results at Month 6. The difference in adjusted mean change in WOMAC Pain Subscale Score from baseline (JTA-200/2 minus reference group) at Month 6 was − 9.49 mm; statistical superiority of JTA-200/2 over the reference was not demonstrated. No statistically significant differences in adjusted mean changes from baseline between JTA-200/2 and reference groups were observed for Pain, Physical Function and Stiffness Subscales WOMAC Scores, Total WOMAC Score, and Well-being Score at any timepoint, although JTA-200/2 induced larger improvements in WOMAC Scores than the reference. Statistically significantly larger improvements in WOMAC Pain Subscale Scores for JTA-004 versus the reference were observed in post-hoc analyses on pooled data from all JTA-004 formulations at Month 6 (p = 0.030) and Month 3 (p = 0.014). All JTA-004 formulations had clinically acceptable safety profiles. Conclusions: This study provided preliminary evidence of the safety of intra-articular injection of JTA-004 in knee osteoarthritis patients. Phase III randomized controlled trials with larger sample sizes are needed to evaluate the efficacy of JTA-004 in knee osteoarthritis. Trial registration: Clinicaltrials.gov/identifier NCT02740231; clinicaltrialsregister.eu/identifier 2015–002117-30. Retrospectively registered 13/4/2016.SCOPUS: ar.jinfo:eu-repo/semantics/publishe

Safety and efficacy of intra-articular injection of JTA-004, a novel viscosupplement, in symptomatic knee osteoarthritis: a randomized, double-blind controlled phase II/III study

Objective: The objective was to assess the safety and efficacy of a single intra-articular administration of JTA-004, a novel viscosupplement, in patients suffering from symptomatic knee osteoarthritis (OA) at 6 months. Design and methods: In this prospective, multicenter, double-blind phase II/III trial (NCT02740231), 164 patients with primary OA knee pain were randomly assigned to one of the three JTA-004 strengths or the comparator treatment (Hylan G-F 20) in a 1:1:1:1 ratio. Safety was assessed by monitoring and reporting vital signs, physical examination, adverse events and concomitant medications. The primary efficacy endpoint was the change from baseline at 6 months in WOMAC® VA3.1 pain subscale. Results: JTA-004 was shown to be well tolerated at all strengths evaluated. At 6 months, patients in the three JTA-004 groups showed a better improvement in pain compared to patients in the comparator group although statistical significance was not achieved. As the three JTA-004 strengths had a similar efficacy, a post hoc analysis was subsequently performed between the pooled JTA-004 treated patients and the comparator group. The exploratory analysis showed a 26.1±2.4 (adjusted mean±SE) mm improvement in pain in the pooled JTA-004 group vs. 15.6±4.1 mm in the comparator group at 6 months, demonstrating a statistically significant superiority of JTA-004 over the comparator (p = 0.030). Conclusions: This study provides first evidences of safety and efficacy of JTA-004 in the treatment of symptomatic knee OA. Efficacy will be further confirmed in a subsequent pivotal Phase III study