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Recombinant adeno-associated virus (rAAV) vector gene delivery systems have demonstrated great promise in clinical trials but continue to face durability and dose-related challenges. Unlike rAAV gene therapy, integrating gene addition approaches can provide curative expression in mitotically active cells and pediatric populations. We explored a novel in vivodelivery approach based on an engineered transposase, Sleeping Beauty(SB100X), delivered as an mRNA within a lipid nanoparticle (LNP), in combination with an rAAV-delivered transposable transgene. This combinatorial approach achieved correction of ornithine transcarbamylase deficiency in the neonatal Spfashmouse model following a single delivery to dividing hepatocytes in the newborn liver. Correction remained stable into adulthood, while a conventional rAAV approach resulted in a return to the disease state. In non-human primates, integration by transposition, mediated by this technology, improved gene expression 10-fold over conventional rAAV-mediated gene transfer while requiring 5-fold less vector. Additionally, integration site analysis confirmed a random profile while specifically targeting TA dinucleotides across the genome. Together, these findings demonstrate that transposable elements can improve rAAV-delivered therapies by lowering the vector dose requirement and associated toxicity while expanding target cell types.

The prodromal stage of Lewy body dementia includes a mild cognitive impairment with visual processing and/or attention-executive deficits. A clinical presentation with progressive visual loss is indeed seldom reported and can be misleading with a posterior cortical atrophy disease. While the neurodegeneration at the occipital cortex can only partially explain the visual disturbances of Lewy body dementia, more recently a retinal dysfunction has been suggested by preliminary optical coherence tomography and autoptic findings. Herein, we present a case of a mild cognitive impairment with Lewy bodies, who presented initially with visual disturbances and signs of both retinal and cortical visual processing dysfunction. A complete neuropsychological, neurophysiological and brain imaging assessment highlighted a prominent ventral visual pathway involvement. This report provides first that the prodromal stage of Lewy body dementia can manifest as a primarily progressive visual loss, second that the involvement of visual pathway, particularly the ventral stream, can be detectable from the retinal to the cortical level.

No study has evaluated the cardiovascular effects of diazepam in elderly subjects that assume diazepam to induce sleep. The present study was carried out in order to evaluate the effects of chronic administration of diazepam as hypnotic drug on blood pressure (BP) and heart rate (HR) in healthy elderly subjects. Healthy, elderly subjects, aged 65–74 years, were treated with diazepam 5 mg or placebo—both administered once a day in the evening—for 4 weeks in two cross-over periods, each separated by a 2-week placebo period, according to a randomized, double-blind, cross-over design. At the end of each study period, clinical as well as 24-h ambulatory BP and HR were evaluated. A total of 25 subjects were included in the analysis. At the end of a 4-week diazepam treatment, clinical as well 24-h BP and HR mean values were not significantly affected. Analysis of sub-periods showed that during night-time, systolic BP (SBP) values under diazepam were 7.6% higher than under placebo, with a mean difference of 7.9 mmHg (p< 0.01), diastolic BP (DBP) values were 5.8% higher, with a mean difference of 3.7 mmHg (p< 0.05 vs placebo) and HR values were 6.6% higher with a mean difference of 4.2 b/min (p< 0.05). The HR increase observed with diazepam persisted during the morning hours, whereas during the afternoon and evening hours SBP, DBP and HR values were similar in the two treatment groups. In elderly subjects chronic assumption of diazepam as hypnotic agent produced an increase in BP, in particular SBP, during night-time and of HR during night-time and morning hours. These effects, which probably depend on a diazepam-mediated increase in sympathetic drive and decrease in vagal tone, might be of clinical relevance due to the role of increased BP and HR as independent predictors of cardiovascular morbidity and mortality.

The FDA approval of autologous Chimeric Antigen Receptor (CAR) T-cell therapies have added a powerful tool in the armamentarium to treat relapsed and refractory leukemias and lymphomas. However, significant challenges limit these therapies from accommodating current patient demand including supply chain limitations and wait times associated with availability of GMP quality viral vectors and long needle-to-needle time from initiation of treatment to receiving the final drug product. As of yet other competitive therapies including bi-specific antibodies and allogeneic CAR-T have not achieved the same level of therapeutic efficacy as conventional autologous T cell therapies highlighting the critical unmet need for same day treatment options for autologous CAR-T products.

The NA48/2 experiment collected data in 2003 and 2004 with the main purpose to study direct CP violation in charged kaons decaying in three pions. Thanks to the huge statistics collected, other studies are also possible. In this paper two recent results, in the framework of QCD studies, are reported: the measurement of the K±→ π0e±νForm Factors and the first observation of the rare decay K±→ π±π0e+e−.

The K+→π+νν‾decay is theoretically one of the cleanest meson decays and so a good place to look for indirect effects of new physics complementary to LHC searches. The NA62 experiment at CERN is designed to measure the branching ratio of this decay with 10% precision. NA62 was commissioned in October 2014, took data in pilot runs in 2014 and 2015. The NA62 experimental setup is illustrated and data quality is reported.

Among the meson decays, K→πνν‾are the cleanest environment, from the theoretical point of view, where to search for new physics effects. The NA62 Experiment at CERN SPS aims to measure the BR(K+→π+νν‾)with a 10% precision by the end of 2018. It has been commissioned with technical runs in 2014 and 2015, and some preliminary results of the detector performances and quality of data are here reported.

The NA62 experiment at CERN collected a large sample of charged kaon decays with a highly efficient trigger for decays into electrons in 2007. The kaon beam represents a source of tagged neutral pion decays in vacuum. A measurement of the electromagnetic transition form factor slope of the neutral pion in the time-like region from ∼1 million fully reconstructed π0Dalitz decay is presented. The limits on dark photon production in π0decays from the earlier kaon experiment at CERN, NA48/2, are also reported.

Effects of new physics in flavor could be found both in Flavor Changing Neutral Current (FCNC) processes and in Lepton Flavor Violation (LFV) modes. The former offer the possibility to deeply test the standard model in a clean environment, while the latter are sensitive to contribution from several models beyond the standard model. In the Kaon sector both FCNC and LFV will be investigated in the NA62 experiment. In addition the kaons sector is an ideal place where to look for new particles and tiny effects, in the region of hundreds of MeV/c2. In this paper prospects for exotic searches in NA62 will be presented, together with recent results from NA48/2 and NA62-RK on LFV kaon decays modes.

The main physics goal of the NA62 experiment at CERN is to precisely measure the branching ratio of the Kaon rare decay K+→π+νν‾. This decay is strongly suppressed in the Standard Model. On the other hand its branching ratio is calculated with high accuracy. NA62 is designed to measure the K+→π+νν‾decay rate with an uncertainty better than 10%. The measurement can serve as a probe to some new physics phenomena, which can alter the decay rate. The NA62 experiment has been successfully launched in October 2014. The theory framework as well as the NA62 detector and the preliminary results are reviewed in this article.

Trigeminal autonomic cephalalgias (TACs) are a group of primary headaches including cluster headache (CH), paroxysmal hemicrania (PH) and short-lasting unilateral neuralgiform headache with conjunctival injection and tearing (SUNCT). Another form, hemicrania continua (HC), is also included this group due to its clinical and pathophysiological similarities. CH is the most common of these syndromes, the others being infrequent in the general population. The pathophysiology of the TACs has been partly elucidated by a number of recent neuroimaging studies, which implicate brain regions associated with nociception (pain matrix). In addition, the hypothalamic activation observed in the course of TAC attacks and the observed efficacy of hypothalamic neurostimulation in CH patients suggest that the hypothalamus is another key structure. Hypothalamic activation may indeed be involved in attack initiation, but it may also lead to a condition of central facilitation underlying the recurrence of pain episodes. The TACs share many pathophysiological features, but are characterised by differences in attack duration and frequency, and to some extent treatment response. Although alternative strategies for the TACs, especially CH, are now emerging (such as neurostimulation techniques), this review focuses on the available pharmacological treatments complying with the most recent guidelines. We discuss the clinical efficacy and tolerability of the currently used drugs. Due to the low frequency of most TACs, few randomised controlled trials have been conducted. The therapies of choice in CH continue to be the triptans and oxygen for acute treatment, and verapamil and lithium for prevention, but promising results have recently been obtained with novel modes of administration of the triptans and other agents, and several other treatments are currently under study. Indomethacin is extremely effective in PH and HC, while antiepileptic drugs (especially lamotrigine) appear to be increasingly useful in SUNCT. We highlight the need for appropriate studies investigating treatments for these rare, but lifelong and disabling conditions.

A mathematically consistent definition of series was derived by the application of Principal Component Analysis to 3-D similarity indices (ASP Software). For two data sets of aromatic molecules, a mono-dimensional numerical ordering was derived, corresponding to the consensus distance from the series lead. The series is unambiguously defined in terms of location along the mono-dimensional axis, along which shape and electrostatic features of the molecules vary in a coordinated way. Molecules along the axis are characterized by the same pattern of electrostatic potential and shape shared by the lead compound, of which they represent linearly “shifted” replicas. This approach can contribute to the exploration of the chemical spaces from a local “fine grain” perspective, thus complementing the “coarse grain” descriptions normally used in the analysis of large data sets.

A mathematically consistent definition of series was derived by the application of Principal Component Analysis to 3-D similarity indices (ASP Software). For two data sets of aromatic molecules, a mono-dimensional numerical ordering was derived, corresponding to the consensus distance from the series lead. The series is unambiguously defined in terms of location along the mono-dimensional axis, along which shape and electrostatic features of the molecules vary in a coordinated way. Molecules along the axis are characterized by the same pattern of electrostatic potential and shape shared by the lead compound, of which they represent linearly “shifted” replicas. This approach can contribute to the exploration of the chemical spaces from a local “fine grain” perspective, thus complementing the “coarse grain” descriptions normally used in the analysis of large data sets.

A versatile new method has been developed as a continuous symmetry measure for chiral compounds. The application of principal component analysis (PCA) to the complete N × N pairwise similarity matrices (electrostatic potential and shape indices) of a series of dihydropyridine calcium channel antagonists allowed to single out a chirality component and to compute a chirality score in terms of the between-enantiomers difference on the component value. The possibility to have chirality defined continuously at the series level could be of importance in eudismic analyses where the relative potency of two enantiomers is studied as well as in QSAR studies dealing with chiral molecules in order to improve the power of the generated models.

A generalization of Wilson loop observables for BFtheories in any dimension is introduced within the Batalin–Vilkovisky framework. The expectation values of these observables are cohomology classes of the space of imbeddings of a circle. One of the resulting theories discussed in the Letter has only trivalent interactions and, irrespective of the actual dimension, looks like a three-dimensional Chern–Simons theory.

Abstract:: We study the differential geometry of principal G-bundles whose base space is the space of free paths (loops) on a manifold M. In particular we consider connections defined in terms of pairs (A,B), where A is a connection for a fixed principal bundle P(M,G) and B is a 2-form on M. The relevant curvatures, parallel transports and holonomies are computed and their expressions in local coordinates are exhibited. When the 2-form B is given by the curvature of A, then the so-called non-abelian Stokes formula follows. For a generic 2-form B, we distinguish the cases when the parallel transport depends on the whole path of paths and when it depends only on the spanned surface. In particular we discuss generalizations of the non-abelian Stokes formula. We study also the invariance properties of the (trace of the) holonomy under suitable transformation groups acting on the pairs (A,B). In this way we are able to define observables for both topological and non-topological quantum field theories of the BF type. In the non-topological case, the surface terms may be relevant for the understanding of the quark-confinement problem. In the topological case the (perturbative) four-dimensional quantum BF-theory is expected to yield invariants of imbedded (or immersed) surfaces in a 4-manifold M.

The results about the isomorphism of a quantum logic L with the logic of the projections in a separable Hilbert space previously obtained with the introduction of the topology of states are completed, including the case of non-separable Hilbert space, and showing that the continuity of the antiautomorphism ? of the division ring R, C or Q determined by L follows from the general topological assumptions on L.

First we briefly recall the definition of the three-dimensional Baxter-Bazhanov lattice model. The spins of this model are elements ofZN and theR-matrix is associated to the algebraUqsl(n) ifq is a primitiveNth root of unity. Then we construct a particularN→∞ limit of the model, in which it is meaningful to interpret the spins as elements ofR and which gives the free Gaussian boson model. Finally, we study special limits of the rapidity variables in which we obtain braid group representations and we show that forn odd the associated knot invariants are given by the inverse of products of Alexander polynomials, evaluated at certain roots of unity.

Inclusion complex formation between pyridoxine and β- and γ-cyclodextrins in aqueous solution has been investigated by1H NMR and UV spectroscopy. Both complexes exhibited a 1:1 stoichiometry and the inclusion process has been shown to perturb the equilibrium between the lactim and the lactam tautomer of pyridoxine, with a preferential inclusion of the former, less polar tautomer.

In a previous article, we demonstrated that the structure–activity relationship model for the mutagenic potency of aromatic amines is different from that for discriminating between mutagens and nonmutagens. In this work, we present further analyses on the molecular determinants of the mutagenicity of aromatic amines. Based on the use of various methodological approaches, our results indicate that mutagenic activity is influenced by different molecular characteristics in different subclasses of aromatic amines. Thus, the general lesson of this article is that 1) in genetic toxicology, it is necessary to separately investigate the structure–activity relationships for discrimination between positive and negative chemicals, and the structure–activity relationships for the potency of the positive chemicals; 2) in structure–activity studies, it is necessary to investigate the degree of homogeneity (congenericity) of apparently similar chemicals in order to assess and describe the various mechanisms of action that may be elicited by the chemicals. Environ. Mol. Mutagen. 32:75–83, 1998 © 1998 Wiley-Liss, Inc.

Abstract:The classical action for pure Yang-Mills gauge theory can be formulated as a deformation of the topological BF theory where, beside the two-form field B, one has to add one extra-field η given by a one-form which transforms as the difference of two connections. The ensuing action functional gives a theory that is both classically and quantistically equivalent to the original Yang-Mills theory. In order to prove such an equivalence, it is shown that the dependency on the field η can be gauged away completely. This gives rise to a field theory that, for this reason, can be considered as semi-topological or topological in some but not all the fields of the theory. The symmetry group involved in this theory is an affine extension of the tangent gauge group acting on the tangent bundle of the space of connections. A mathematical analysis of this group action and of the relevant BRST complex is discussed in detail.

A prototype 64-channel Fastbus TDC built at Fermilab is described. The module features a full custom CMOS four-channel gated integrator chip. One level of analog buffering at the inputs is implemented on chip. A four-event-deep output queue at the bus interface allows a high event rate with low dead time. Each channel can record up to two hits per event. With an occupation rate of 10%, the module can operate at 40000 events/s with dead time of the order of 15%. The TDC operates in common stop mode with a full scale of 1 mu s and a resolution of 1 ns. Data compaction and a leading word count format have been implemented. The board is less complex than current designs.<>

Summary The safety and efficacy of a single daily dose of fenofibrate (200 mg) have been evaluated in 12 Type IIB hyperlipidaemic patients in a three-month study. At the same time the pharmacokinetics was studied to check whether this new dosage schedule would give a therapeutic plasma levels of fenofibrate.

We show that ghosts in gauge theories can be interpreted as Maurer-Cartan forms in the infinite dimensional group g of gauge transformations. We examine the cohomology of the Lie algebra of g and identify the coboundary operator with the BRS operator. We describe the anomalous terms encountered in the renormalization of gauge theories (triangle anomalies) as elements of these cohomology groups.

We introduce for lattice gauge theories an analogue of the Pontrjagin index and a notion of “selfduality” and “antiselfduality”. Selfdual and antiselfdual configurations on the lattice have much of the same properties (with some remarkable differences) as the corresponding configurations on the continuum, to which they converge when the lattice spacing goes to zero.

We apply the Faddeev-Reshetikhin-Taktajan method for the construction of Quantum Groups to the Yang-Baxter matrices which are related to the invariants of oriented links in Σ×[0,1], where Σ is a non-trivial 2-dimensional surface. We obtain multi-parameter ribbon Hopf algebras that differ in many respects from their one-parameter counterparts. Among the main differences we mention the existence of a non-central quantum determinant and the fact that the number of independent generators is higher than in the one-parameter case.

The rôle of the evaluation map in anomaly calculations for field theory, sigma-models and strings is investigated. In this paper, anomalies in field theory (with and without a backgrounds connection), are obtained as pull-backs of suitable forms via evaluation maps. The cohomology of the group of gauge transformations is computed in terms of the cohomology of the base manifold and of the cohomology of the structure group. This allows us to clarify the different “topological significance” of gauge and gravitational anomalies. The relation between “locality” and “universality” is discussed and “local cohomology” is linked to the cohomology of classifying spaces. The problem of combining the locality requirement and the index theorem approach to anomalies is also examined. Anomaly cancellation in field theories derived from superstrings is analyzed and the relevant geometrical constraints are discussed.

A pair of small angle silicon-tungsten (Si-W) calorimeters has been built to measure the luminosity to a precision better than 0.1% in the OPAL experiment at the Large Electron Positron (LEP) collider at CERN near Geneva. Each calorimeter contains 19 layers of tungsten (W) plates and silicon (Si) detectors, corresponding to a total of 22 radiation lengths, sampled by about 1 m/sup 2/ of detectors divided into 304/spl times/64 independently read out channels. A complete electronics system has been developed, from the preamplifier up to the VME read out and control interface. It includes a fast trigger based on analogue sums. This paper describes how a large number of channels have been implemented in a dense environment, thanks to the use of ASIC's directly bonded on the detector.<>

Following and generalizing a paper by Turaev, we consider some algebraic structures on the set of (generalized) link-diagrams, meant simply as collections of immersed loops on a two dimensional surface, with the specification of an over/undercrossing prescription at each double point. This definition is general enough to be relevant not only to traditional knot-theory, but possibly to some statistical mechanical models. A coalgebra structure is introduced on some modules (over polynomial rings), generated by these diagrams. The compatibility of this coalgebra structure with the skein invariance and the invariance under Reidemeister moves is discussed. A Hopf algebra structure results only in some special cases, which are thoroughly examined. It is shown that a special choice of the ground (polynomial) ring over which the diagram module is defined, allows us to define link-invariants for links (in the ordinary sense) in Σ×[0,1],where Σ is a (closed or open) two-dimensional surface. These invariants generalize in a nontrivial way the Jones polynomials and the Homfly polynomials (at least when the last ones are computed for some special values of the variables). In a sequel paper the relation between the algebraic structures of link-diagrams, some special types of quantum groups and the quantum holonomy will be discussed.

In a previous article, we demonstrated that the structure–activity relationship model for the mutagenic potency of aromatic amines is different from that for discriminating between mutagens and nonmutagens. In this work, we present further analyses on the molecular determinants of the mutagenicity of aromatic amines. Based on the use of various methodological approaches, our results indicate that mutagenic activity is influenced by different molecular characteristics in different subclasses of aromatic amines. Thus, the general lesson of this article is that 1) in genetic toxicology, it is necessary to separately investigate the structure–activity relationships for discrimination between positive and negative chemicals, and the structure–activity relationships for the potency of the positive chemicals; 2) in structure–activity studies, it is necessary to investigate the degree of homogeneity (congenericity) of apparently similar chemicals in order to assess and describe the various mechanisms of action that may be elicited by the chemicals. Environ. Mol. Mutagen. 32:75–83, 1998 © 1998 Wiley‐Liss, Inc.

In this paper, we examine specifically the rôle of the evaluation map in sigma-models and strings. We discuss the difference between sigma-models and field theory, as far as anomaly cancellation is concerned. The introduction of the Wess—Zumino terms in different sigma-models is considered. Anomalies in string theory are discussed, with special attention to the conformal anomalies and to the sigma-model anomalies for the imbedded (or immersed) world-sheet of the string. Conformal anomalies in two dimensions are connected to holomorphic and gravitational anomalies. In order to have the cancellation of the sigma-model anomalies of the string, certain topological conditions must be satisfied by the ambient manifold. The rôle of the evaluation map in the calculations of global anomalies is also discussed, both for field theories and for sigma-models. In particular global anomalies are connected with the differential characters of Cheeger and Simons. We show that the absence of global anomalies in sigma-models is guaranteed by the absence of torsion in suitable homology groups of the target space.

A topology is introduced in a logic L using the set of pure states of L. It is shown that L, equipped with this topology, under suitable conditions, determines the division ring R, C or 2e. With the continuity of the antiautomorphism of the division ring added, it is shown that these conditions are necessary and sufficient for the projective logic L to be isomorphic with the projective logic of the projections in a Hilbert space over R, C or 2e.

In this paper the pretrigger system designed for the electromagnetic calorimeter of the HERA-B experiment is described. This system will provide the electron candidates to the first-level trigger. The first results obtained with a whole readout and pretrigger prototype chain are also described.

Introduction:prevalence of insomnia increases with age and benzodiazepines are the most widely prescribed hypnotic drugs. However, no study to date has evaluated their cardiovascular effects in hypertensive patients.Hypothesis:to evaluate whether the evening chronic assumption of bromazepam, or alprazolam or lorazepam to induce sleep has any effect on blood pressure (BP) and heart rate (HR) in mild hypertensive patients.Methods:one hundred and twenty new diagnosed never treated mild hypertensive patients aged 40-65 years, after a 2 week placebo period were randomized to bromazepam 3 mg or alprazolam 0.5 mg or lorazepam 1 mg. The drugs were taken orally in the evening between 10.00 and 11.00 pm for 4 weeks according to a double-blind cross-over design. At the end of each treatment period 24-hour ambulatory BP and HR were evaluated.Results:24-hour ambulatory BP, was not significantly affected by any treatment; however a small but significant increase (+ 3.5%) nighttime DBP was detected with the lorazepam treatment. Nighttime HR was significantly higher with bromazepam (+ 11.5%) and with lorazepm (+ 9.7%). No change in BP and HR was detected with alprazolam.Conclusions:in mild untreated hypertensive patients chronic assumption of alproazolam as a hypnotic agent, is absolutely devoid of any effect on nocturnal BP and HR. On the opposite bromazepam induces increase in HR and lorazepam an increase of both HR and DBP. This might be of clinical relevance due to the role of increased HR and BP as independent cardiovascular risk factor.

We have previously shown that CRISPR/Cas9 mediates high levels of gene editing in adult human CD34+hematopoietic stem and progenitor cells (HSPCs). We now demonstrated that high levels of gene editing can also be achieved in HSPCs using Cpf1. In addition to NHEJ-mediated gene editing, we explored targeted integration at the ß-hemoglobin gene (HBB) with Cas9, toward gene correction for sickle cell disease (SCD). For targeted integration strategies at HBB, we designed AAV6 DNA donor repair templates flanked by either DNA sequences homologous to the cut site in the genomic location of the SCD mutation or DNA sequences non-homologous to the human genome. To evaluate targeted integration in HSPCs, adult mobilized peripheral blood (mPB) CD34+cells were electroporated with Cas9 ribonucleoprotein (RNP) and were then transduced with AAV6 containing DNA repair templates with or without homology arms. We analyzed targeted integration at the HBBtarget site by digital droplet PCR (ddPCR) analysis 7 days after transduction of mPB CD34+cells. The ddPCR results show ~30% targeted integration in HBBin mPB CD34+cells with no impact on cell viability. DNA sequencing analysis supported the ddPCR results indicating that perfect homology directed repair-driven integration of the DNA donor template was the preferential repair mechanism. Less than 1% of targeted integration was detected by ddPCR in mPB CD34+cells transduced with AAV6 DNA repair template that lacked homology arms, suggesting that the mechanism of integration is homology repair dependent. These integrations were maintained in the Glycophorin A+erythroid progeny of genome-edited mPB CD34+cells, paving the way toward gene correction for SCD. The combination of CRISPR nucleases (Cpf1, Cas9) and AAV6 DNA repair templates form a powerful genome editing platform that enables broader applications for the treatment of hematologic diseases beyond hemoglobinopathies.

Transplantation of gene-modified autologous hematopoietic stem/progenitor cells (HSPCs) is an effective treatment for several hematologic diseases. However, a number of blood disorders may not be amenable to gene augmentation-based therapeutics. Targeted genome editing in human HSPCs could provide a therapeutic approach for these otherwise untreatable diseases. Here we demonstrate that CRISPR/Cas9 ribonucleoprotein (RNP) edits target genes in human HSPCs with high efficiency and precision. Human adult and umbilical cord blood (CB) CD34+cells from 20 donors were electroporated with S. pyogenesor S. aureusCas9 RNP targeting HBB, AAVS1, or CXCR4. Sequence analysis demonstrated up to 80% editing in CB CD34+cells (mean±s.d: 61%±9%) and up to 57% in adult CD34+cells (39%±13%). Delivery of Cas9 RNP and a single-stranded oligodeoxynucleotide donor (ssODN) led to up to 12% ssODN-mediated homology directed repair (HDR) and also led to a 20% increase in total gene editing (HDR+NHEJ)(RNP: 48%±15%; RNP+ssODN: 69%±8%). Both Cas9 RNP gene-edited CD34+cells and donor-matched untreated control CD34+cells reconstituted human hematopoiesis in primary and secondary recipient immunodeficient mice, with ~85% human CD45+cell peripheral blood reconstitution 4 months after primary transplantation. Human T and B lymphoid, erythroid, and myeloid cells were detected in the spleen, thymus, and bone marrow with 20% CD34+cell engraftment in the marrow of mice transplanted with RNP gene-edited or control CD34+cells. The level of targeted gene editing in human erythroid, myeloid, and CD34+cells that were recovered and enriched from the hematopoietic organs of primary recipients (~50%) was similar to the level of gene editing detected in the pre-infusion product (~60%). In summary, these results indicate that Cas9 gene-edited human HSPCs retain long-term engraftment potential and support multilineage blood reconstitution in vivo, thus supporting further investigation of CRISPR/Cas9 mediated gene-edited hematopoietic stem/progenitor cell therapies.

Heath: Editas Medicine: Employment. Chalishazar:Editas Medicine: Employment. Lee:Editas Medicine: Employment. Selleck:Editas Medicine: Employment. Cotta-Ramusino:Editas Medicine: Employment. Bumcrot:Editas Medicine: Employment. Gori:Editas Medicine: Employment.