Physiological aging and inflammation-induced cellular senescence may contribute to oligodendroglial dysfunction in MS.

Aging affects all cell types in the CNS and plays an important role in CNS diseases. However, the underlying molecular mechanisms driving these age-associated changes and their contribution to diseases are only poorly understood. The white matter in the aging brain as well as in diseases, such as Multiple sclerosis is characterized by subtle abnormalities in myelin sheaths and paranodes, suggesting that oligodendrocytes, the myelin-maintaining cells of the CNS, lose the capacity to preserve a proper myelin structure and potentially function in age and certain diseases. Here, we made use of directly converted oligodendrocytes (dchiOL) from young, adult and old human donors to study age-associated changes. dchiOL from all three age groups differentiated in an comparable manner into O4 + immature oligodendrocytes, but the proportion of MBP + mature dchiOL decreased with increasing donor age. This was associated with an increased ROS production and upregulation of cellular senescence markers such as CDKN1A, CDKN2A in old dchiOL. Comparison of the transcriptomic profiles of dchiOL from adult and old donors revealed 1324 differentially regulated genes with limited overlap with transcriptomic profiles of the donors' fibroblasts or published data sets from directly converted human neurons or primary rodent oligodendroglial lineage cells. Methylome analyses of dchiOL and human white matter tissue samples demonstrate that chronological and epigenetic age correlate in CNS white matter as well as in dchiOL and resulted in the identification of an age-specific epigenetic signature. Furthermore, we observed an accelerated epigenetic aging of the myelinated, normal appearing white matter of multiple sclerosis (MS) patients compared to healthy individuals. Impaired differentiation and upregulation of cellular senescence markers could be induced in young dchiOL in vitro using supernatants from pro-inflammatory microglia. In summary, our data suggest that physiological aging as well as inflammation-induced cellular senescence contribute to oligodendroglial pathology in inflammatory demyelinating diseases such as MS.

The efficient induction of human retinal ganglion-like cells provides a platform for studying optic neuropathies.

Retinal ganglion cells (RGCs) are essential for vision perception. In glaucoma and other optic neuropathies, RGCs and their optic axons undergo degenerative change and cell death; this can result in irreversible vision loss. Here we developed a rapid protocol for directly inducing RGC differentiation from human induced pluripotent stem cells (hiPSCs) by the overexpression of ATOH7, BRN3B, and SOX4. The hiPSC-derived RGC-like cells (iRGCs) show robust expression of various RGC-specific markers by whole transcriptome profiling. A functional assessment was also carried out and this demonstrated that these iRGCs display stimulus-induced neuronal activity, as well as spontaneous neuronal activity. Ethambutol (EMB), an effective first-line anti-tuberculosis agent, is known to cause serious visual impairment and irreversible vision loss due to the RGC degeneration in a significant number of treated patients. Using our iRGCs, EMB was found to induce significant dose-dependent and time-dependent increases in cell death and neurite degeneration. Western blot analysis revealed that the expression levels of p62 and LC3-II were upregulated, and further investigations revealed that EMB caused a blockade of lysosome-autophagosome fusion; this indicates that impairment of autophagic flux is one of the adverse effects of that EMB has on iRGCs. In addition, EMB was found to elevate intracellular reactive oxygen species (ROS) levels increasing apoptotic cell death. This could be partially rescued by the co-treatment with the ROS scavenger NAC. Taken together, our findings suggest that this iRGC model, which achieves both high yield and high purity, is suitable for investigating optic neuropathies, as well as being useful when searching for potential drugs for therapeutic treatment and/or disease prevention.

Transplantation of Chemical Compound-Induced Cells from Human Fibroblasts Improves Locomotor Recovery in a Spinal Cord Injury Rat Model.

The development of regenerative medicine using cell therapy is eagerly awaited for diseases such as spinal cord injury (SCI), for which there has been no radical cure. We previously reported the direct conversion of human fibroblasts into neuronal-like cells using only chemical compounds; however, it is unclear whether chemical compound-induced neuronal-like (CiN) cells are clinically functional. In this study, we partially modified the method of inducing CiN cells (termed immature CiN cells) and examined their therapeutic efficacy, in a rat model of SCI, to investigate whether immature CiN cells are promising for clinical applications. Motor function recovery, after SCI, was assessed using the Basso, Beattie, and Bresnahan (BBB) test, as well as the CatWalk analysis. We found that locomotor recovery, after SCI in the immature CiN cell-transplanted group, was partially improved compared to that in the control group. Consistent with these results, magnetic resonance imaging (MRI) and histopathological analyses revealed that nerve recovery or preservation improved in the immature CiN cell-transplanted group. Furthermore, transcriptome analysis revealed that immature CiN cells highly express hepatocyte growth factor (HGF), which has recently been shown to be a promising therapeutic agent against SCI. Our findings suggest that immature CiN cells may provide an alternative strategy for the regenerative therapy of SCI.

Direct Conversion of Cell Fate and Induced Endothelial Cells.

Advances in nuclear reprogramming technology have enabled the dedifferentiation and transdifferentiation of mammalian cells. Forced induction of the key transcription factors constituting a transcriptional network can convert cells back to their pluripotent status or directly to another cell fate without inducing pluripotency. To date, direct conversion to several cell types, including cardiomyocytes, various types of neurons, and pancreatic ß-cells, has been reported. We previously demonstrated direct lineage reprogramming of adult fibroblasts into induced endothelial cells (iECs) in mice and humans. In contrast to induced pluripotent stem cells, for which there is consensus on the criteria defining pluripotency, such criteria have not yet been established in the field of direct conversion. We thus suggest that careful assessment of the status of converted cells using genetic and epigenetic profiling, various functional assays, and the use of multiple readouts is essential to determine successful conversion. As direct conversion does not go through pluripotent status, this technique can be utilized for therapeutic purposes without the risk of tumorigenesis. Further, direct conversion can be induced in vivo by gene delivery to the target tissue or organ in situ. Thus, direct conversion technology can be developed into cell therapy or gene therapy for regenerative purposes. Here, we review the potential and future directions of direct cell fate conversion and iECs.

Application of Approximation Constructions with a Small Number of Parameters for the Estimation of a Rayleigh Fading Multipath Channel with Doppler Spectrum Spreading.

In this article, an algorithm for joint estimation of communication channel gains and signal distortions in a direct conversion receiver is proposed. The received signal model uses approximations with a small number of parameters to reduce the computational complexity of the resulting algorithm. The estimation algorithm is obtained under the assumption of a priori uncertainty about the characteristics of the communication channel and noise distribution using the linear least squares method. Estimation is performed first by the test sequence, then by the information symbols obtained after detection. In addition, an analysis of the noise immunity of quadrature amplitude modulation (QAM) signal reception is carried out using different approximating structures in the estimation algorithm for systems with a single transmitting and receiving antenna (SISO) and for systems with multiple transmitting and receiving antennas (MIMO). Furthermore, this article examines the influence of the duration of the test signal, the number of sessions of its transmission, and the channel extrapolation interval on the quality of signal reception.

Comparative Analysis of Multilayer Lead Oxide-Based X-ray Detector Prototypes.

Lead oxide (PbO) photoconductors are proposed as X-ray-to-charge transducers for the next generation of direct conversion digital X-ray detectors. Optimized PbO-based detectors have potential for utilization in high-energy and dynamic applications of medical X-ray imaging. Two polymorphs of PbO have been considered so far for imaging applications: polycrystalline lead oxide (poly-PbO) and amorphous lead oxide (a-PbO). Here, we provide the comparative analysis of two PbO-based single-pixel X-ray detector prototypes: one prototype employs only a layer of a-PbO as the photoconductor while the other has a combination of a-PbO and poly-PbO, forming a photoconductive bilayer structure of the same overall thickness as in the first prototype. We characterize the performance of these prototypes in terms of electron-hole creation energy (W±) and signal lag-major properties that define a material's suitability for low-dose real-time imaging. The results demonstrate that both X-ray photoconductive structures have an adequate temporal response suitable for real-time X-ray imaging, combined with high intrinsic sensitivity. These results are discussed in the context of structural and morphological properties of PbO to better understand the preparation-fabrication-property relationships of this material.

A 24 GHz CMOS Direct-Conversion RF Receiver with I/Q Mismatch Calibration for Radar Sensor Applications.

A 24 GHz millimeter-wave direct-conversion radio-frequency (RF) receiver with wide-range and precise I/Q mismatch calibration is designed in 65 nm CMOS technology for radar sensor applications. The CMOS RF receiver is based on a quadrature direct-conversion architecture. Analytic relations are derived to clearly exhibit the individual contributions of the I/Q amplitude and phase mismatches to the image-rejection ratio (IRR) degradation, which provides a useful design guide for determining the range and resolution of the I/Q mismatch calibration circuit. The designed CMOS RF receiver comprises a low-noise amplifier, quadrature down-conversion mixer, baseband amplifier, and quadrature LO generator. Controlling the individual gate bias voltages of the switching FETs in the down-conversion mixer having a resistive load is found to induce significant changes at the amplitude and phase of the output signal. In the calibration process, the mixer gate bias tuning is first performed for the amplitude mismatch calibration, and the remaining phase mismatch is then calibrated out by the varactor capacitance tuning at the LO buffer's LC load. Implemented in 65 nm CMOS process, the RF receiver achieves 31.5 dB power gain, -35.2 dBm input-referred 1 dB compression power, and 4.8-7.1 dB noise figure across 22.5-26.1 GHz band, while dissipating 106.2 mA from a 1.2 V supply. The effectiveness of the proposed I/Q mismatch calibration is successfully verified by observing that the amplitude and phase mismatches are improved from 1.0-1.5 dB to 0.02-0.19 dB, and from 10.8-23.8 to 1.1-3.2 degrees, respectively.

Dark Current Modeling for a Polyimide-Amorphous Lead Oxide-Based Direct Conversion X-ray Detector.

The reduction of the dark current (DC) to a tolerable level in amorphous selenium (a-Se) X-ray photoconductors was one of the key factors that led to the successful commercialization of a-Se-based direct conversion flat panel X-ray imagers (FPXIs) and their widespread clinical use. Here, we discuss the origin of DC in another X-ray photoconductive structure that utilizes amorphous lead oxide (a-PbO) as an X-ray-to-charge transducer and polyimide (PI) as a blocking layer. The transient DC in a PI/a-PbO detector is measured at different applied electric fields (5-20 V/µm). The experimental results are used to develop a theoretical model describing the electric field-dependent transient behavior of DC. The results of the DC kinetics modeling show that the DC, shortly after the bias application, is primarily controlled by the injection of holes from the positively biased electrode and gradually decays with time to a steady-state value. DC decays by the overarching mechanism of an electric field redistribution, caused by the accumulation of trapped holes in deep localized states within the bulk of PI. Thermal generation and subsequent multiple-trapping (MT) controlled transport of holes within the a-PbO layer governs the steady-state value at all the applied fields investigated here, except for the largest applied field of 20 V/µm. This suggests that a thicker layer of PI would be more optimal to suppress DC in the PI/a-PbO detector presented here. The model can be used to find an approximate optimal thickness of PI for future iterations of PI/a-PbO detectors without the need for time and labor-intensive experimental trial and error. In addition, we show that accounting for the field-induced charge carrier release from traps, enhanced by charge hopping transitions between the traps, yields an excellent fit between the experimental and simulated results, thus, clarifying the dynamic process of reaching a steady-state occupancy level of the deep localized states in the PI. Practically, the electric field redistribution causes the internal field to increase in magnitude in the a-PbO layer, thus improving charge collection efficiency and temporal performance over time, as confirmed by experimental results. The electric field redistribution can be implemented as a warm-up time for a-PbO-based detectors.

Millimeter-Wave Imaging System Based on Direct-Conversion Focal-Plane Array Receiver.

A new approach to millimeter-wave imaging was suggested and experimentally studied. This approach can be considered as the evolution of the well-established focal-plane array (FPA) millimeter-wave imaging. The significant difference is the use of a direct-conversion array receiver, instead of the direct-detection array receiver, along with the frequency-modulated continuous-wave (FMCW) radar technique. The sensitivity of the direct-conversion receiver is several orders higher than the sensitivity of the direct-detection one, which allows us to increase the maximum imaging range by more than one order of magnitude. The additional advantage of the direct-conversion technique is the opportunity to obtain information about the range to an object. The realization of the direct-conversion FPA imaging system was made possible due to original sensitive simple-designed receiving elements based on low-barrier Mott diodes. The suggested imaging method's main characteristics, which include the achievable angular and range resolution and the achievable maximum imaging range, were studied. A maximum range of up to 100 m was experimentally determined. A 94 GHz 8 × 8 imaging system was developed for demonstration purposes and studied in detail. The suggested technique is assumed to be useful for creating a long-range millimeter-wave camera, in particular, for robotic systems that operate in poor environmental conditions.

Direct Conversion X-ray Detector with Micron-Scale Pixel Pitch for Edge-Illumination and Propagation-Based X-ray Phase-Contrast Imaging.

In this work, we investigate the potential of employing a direct conversion integration mode X-ray detector with micron-scale pixels in two different X-ray phase-contrast imaging (XPCi) configurations, propagation-based (PB) and edge illumination (EI). Both PB-XPCi and EI-XPCi implementations are evaluated through a wave optics model-numerically simulated in MATLAB-and are compared based on their contrast, edge-enhancement, visibility, and dose efficiency characteristics. The EI-XPCi configuration, in general, demonstrates higher performance compared to PB-XPCi, considering a setup with the same X-ray source and detector. However, absorption masks quality (thickness of X-ray absorption material) and environmental vibration effect are two potential challenges for EI-XPCi employing a detector with micron-scale pixels. Simulation results confirm that the behavior of an EI-XPCi system employing a high-resolution detector is susceptible to its absorption masks thickness and misalignment. This work demonstrates the potential and feasibility of employing a high-resolution direct conversion detector for phase-contrast imaging applications where higher dose efficiency, higher contrast images, and a more compact imaging system are of interest.

The application of in vitro-derived human neurons in neurodegenerative disease modeling.

The development of safe and effective treatments for age-associated neurodegenerative disorders is an on-going challenge faced by the scientific field. Key to the development of such therapies is the appropriate selection of modeling systems in which to investigate disease mechanisms and to test candidate interventions. There are unique challenges in the development of representative laboratory models of neurodegenerative diseases, including the complexity of the human brain, the cumulative and variable contributions of genetic and environmental factors over the course of a lifetime, inability to culture human primary neurons, and critical central nervous system differences between small animal models and humans. While traditional rodent models have advanced our understanding of neurodegenerative disease mechanisms, key divergences such as the species-specific genetic background can limit the application of animal models in many cases. Here we review in vitro human neuronal systems that employ stem cell and reprogramming technology and their application to a range of neurodegenerative diseases. Specifically, we compare human-induced pluripotent stem cell-derived neurons to directly converted, or transdifferentiated, induced neurons, as both model systems can take advantage of patient-derived human tissue to produce neurons in culture. We present recent technical developments using these two modeling systems, as well as current limitations to these systems, with the aim of advancing investigation of neuropathogenic mechanisms using these models.

Enhanced Conversion Efficiency Enabled by Species Migration in Direct Solar Energy Storage.

Solar energy can be stored via either an indirect route in which electricity is involved as an intermediate step, or a direct route that utilizes photogenerated charge carriers for direct solar energy conversion. In this study, we investigate the fundamental difference between the direct and indirect routes in solar energy conversion using a new photoelectrochemical energy storage cell (PESC) as a model device. This PESC centers on a liquid junction that utilizes CH3 NH3 PbI3 perovskite to drive photoelectrochemical reactions of Benzoquinone (BQ) and Ferrocene (Fc) redox species. The experimental studies show that the equilibrium redox potentials are 0.1 V and -0.78 V (vs Ag/AgNO3 ) for Fc+ /Fc and BQ/BQ.- , respectively, which would produce a theoretical open-circuit voltage of 0.88 V for the storage device. The physics-based computational analysis shows a relatively flat reaction rate distribution in the electrode for the indirect route; however, in the direct route the photoelectrochemical reaction rate is critically affected by electron concentration due to strong light absorption of the perovskite material, which has been shown to vary by at least 10-fold in the transverse direction across the photoelectrode. The drastic variation of reaction rate in the photoelectrode creates an electric field that is 7.5 times stronger than the bulk electrolyte, which causes the photo-converted reaction product (i. e., BQ.- ) to drift away from the photoelectrode thereby creating a constant reaction driving force. As a result, it has been shown that the intrinsic solar to chemical conversion (ISTC) efficiency improves by ∼40 % for the direct route compared to the indirect route at 0.05 mA/cm2 .

Deubiquitylating enzymes as cancer stem cell therapeutics.

The focus of basic and applied research on core stem cell transcription factors has paved the way to initial delineation of their characteristics, their regulatory mechanisms, and the applicability of their regulatory proteins for protein-induced pluripotent stem cells (protein-IPSC) generation and in further clinical settings. Striking parallels have been observed between cancer stem cells (CSCs) and stem cells. For the maintenance of stem cells and CSC pluripotency and differentiation, post translational modifications (i.e., ubiquitylation and deubiquitylation) are tightly regulated, as these modifications result in a variety of stem cell fates. The identification of deubiquitylating enzymes (DUBs) involved in the regulation of core stem cell transcription factors and CSC-related proteins might contribute to providing novel insights into the implications of DUB regulatory mechanisms for governing cellular reprogramming and carcinogenesis. Moreover, we propose the novel possibility of applying DUBs coupled with core transcription factors to improve protein-iPSC generation efficiency. Additionally, this review article further illustrates the potential of applying DUB inhibitors as a novel therapeutic intervention for targeting CSCs. Thus, defining DUBs as core pharmacological targets implies that future endeavors to develop their inhibitors may revolutionize our ability to regulate stem cell maintenance and differentiation, somatic cell reprogramming, and cancer stem cells.

Transdifferentiation of human adult peripheral blood T cells into neurons.

Human cell models for disease based on induced pluripotent stem (iPS) cells have proven to be powerful new assets for investigating disease mechanisms. New insights have been obtained studying single mutations using isogenic controls generated by gene targeting. Modeling complex, multigenetic traits using patient-derived iPS cells is much more challenging due to line-to-line variability and technical limitations of scaling to dozens or more patients. Induced neuronal (iN) cells reprogrammed directly from dermal fibroblasts or urinary epithelia could be obtained from many donors, but such donor cells are heterogeneous, show interindividual variability, and must be extensively expanded, which can introduce random mutations. Moreover, derivation of dermal fibroblasts requires invasive biopsies. Here we show that human adult peripheral blood mononuclear cells, as well as defined purified T lymphocytes, can be directly converted into fully functional iN cells, demonstrating that terminally differentiated human cells can be efficiently transdifferentiated into a distantly related lineage. T cell-derived iN cells, generated by nonintegrating gene delivery, showed stereotypical neuronal morphologies and expressed multiple pan-neuronal markers, fired action potentials, and were able to form functional synapses. These cells were stable in the absence of exogenous reprogramming factors. Small molecule addition and optimized culture systems have yielded conversion efficiencies of up to 6.2%, resulting in the generation of >50,000 iN cells from 1 mL of peripheral blood in a single step without the need for initial expansion. Thus, our method allows the generation of sufficient neurons for experimental interrogation from a defined, homogeneous, and readily accessible donor cell population.

The X-ray Sensitivity of an Amorphous Lead Oxide Photoconductor.

The photoconductor layer is an important component of direct conversion flat panel X-ray imagers (FPXI); thus, it should be carefully selected to meet the requirements for the X-ray imaging detector, and its properties should be clearly understood to develop the most optimal detector design. Currently, amorphous selenium (a-Se) is the only photoconductor utilized in commercial direct conversion FPXIs for low-energy mammographic imaging, but it is not practically feasible for higher-energy diagnostic imaging. Amorphous lead oxide (a-PbO) photoconductor is considered as a replacement to a-Se in radiography, fluoroscopy, and tomosynthesis applications. In this work, we investigated the X-ray sensitivity of a-PbO, one of the most important parameters for X-ray photoconductors, and examined the underlying mechanisms responsible for charge generation and recombination. The X-ray sensitivity in terms of electron-hole pair creation energy, W±, was measured in a range of electric fields, X-ray energies, and exposure levels. W± decreases with the electric field and X-ray energy, saturating at 18-31 eV/ehp, depending on the energy of X-rays, but increases with the exposure rate. The peculiar dependencies of W± on these parameters lead to a conclusion that, at electric fields relevant to detector operation (~10 V/µm), the columnar recombination and the bulk recombination mechanisms interplay in the a-PbO photoconductor.

Cell-Permeable Oct4 Gene Delivery Enhances Stem Cell-like Properties of Mouse Embryonic Fibroblasts.

Direct conversion of one cell type into another is a trans-differentiation process. Recent advances in fibroblast research revealed that epithelial cells can give rise to fibroblasts by epithelial-mesenchymal transition. Conversely, fibroblasts can also give rise to epithelia by undergoing a mesenchymal to epithelial transition. To elicit stem cell-like properties in fibroblasts, the Oct4 transcription factor acts as a master transcriptional regulator for reprogramming somatic cells. Notably, the production of gene complexes with cell-permeable peptides, such as low-molecular-weight protamine (LMWP), was proposed to induce reprogramming without cytotoxicity and genomic mutation. We designed a complex with non-cytotoxic LMWP to prevent the degradation of Oct4 and revealed that the positively charged cell-permeable LMWP helped condense the size of the Oct4-LMWP complexes (1:5 N:P ratio). When the Oct4-LMWP complex was delivered into mouse embryonic fibroblasts (MEFs), stemness-related gene expression increased while fibroblast intrinsic properties decreased. We believe that the Oct4-LMWP complex developed in this study can be used to reprogram terminally differentiated somatic cells or convert them into stem cell-like cells without risk of cell death, improving the stemness level and stability of existing direct conversion techniques.

Programming of ES cells and reprogramming of fibroblasts into renal lineage-like cells.

This study aims to prepare intermediate mesoderm-like cells from mouse embryonic fibroblasts (MEFs). In the first step, intermediate mesoderm-like cells (IMLCs) and renal epithelial-like cells (RELCs) were extracted from mouse embryonic stem cells (mESCs) in a specified media that contained two small molecules, CHIR99021 and TTNPB, along with growth factors, FGF9and BMP7. Then, MEFs were directly converted into IM by genes for the pluripotency factors, which encode the transcription factors; Oct4, Sox2, Klf4, and c-Myc (OSKM). These unstable intermediate cells were quickly encouraged to form IM with the assistance of CHIR99021 and TTNPB. The results showed that exogenous expression of OSKM factors for four days was adequate to generate partially reprogrammed cells (SSEA1+/Nanog-). Real-time PCR and immunocytochemistry analysis confirmed the presence of the MEF-derived IMs. This study introduced a method for mESCs differentiation to RELCs followed by MEF conversion in an attempt to generate IM by circumventing pluripotency.

Partial reprogramming as a therapeutic approach for heart disease: A state-of-the-art review.

Heart disease such as myocardial infarction is the first cause of mortality in all countries. Today, cardiac cell-based therapy using de novo produced cardiac cells is considered as a novel approach for cardiac regenerative medicine. Recently, an alchemy-like approach, known as direct reprogramming or direct conversion, has been developed to directly convert somatic cells to cardiac cells in vitro and in vivo. This cellular alchemy is a short-cut and safe strategy for generating autologous cardiac cells, and it can be accomplished through activating cardiogenesis- or pluripotency-related factors in noncardiac cells. Importantly, pluripotency factors-based direct cardiac conversion, known as partial reprogramming, is shorter and more efficient for cardiomyocyte generation in vitro. Today, this strategy is achievable for direct conversion of mouse and human somatic cells to cardiac lineage cells (cardiomyocytes and cardiac progenitor cells), using transgene free, chemical-based approaches. Although, heart-specific partial reprogramming seems to be challenging for in vivo conversion of cardiac fibroblasts to cardiac cells, but whole organism-based in vivo partial reprogramming ameliorates cellular and physiological hallmarks of aging and prolongs lifespan in mouse. Notably, cardiac cells produced using partial reprogramming strategy can be a useful platform for disease modeling, drug screening and cardiac cell-based therapy, once the safety issues are overcome. Herein, we discuss about all progresses in de novo production of cardiac cells using partial reprogramming-based direct conversion, as well as give an overview about the potential applications of this strategy in vivo and in vitro.

Oct4-induced oligodendrocyte progenitor cells enhance functional recovery in spinal cord injury model.

The generation of patient-specific oligodendrocyte progenitor cells (OPCs) holds great potential as an expandable cell source for cell replacement therapy as well as drug screening in spinal cord injury or demyelinating diseases. Here, we demonstrate that induced OPCs (iOPCs) can be directly derived from adult mouse fibroblasts by Oct4-mediated direct reprogramming, using anchorage-independent growth to ensure high purity. Homogeneous iOPCs exhibit typical small-bipolar morphology, maintain their self-renewal capacity and OPC marker expression for more than 31 passages, share high similarity in the global gene expression profile to wild-type OPCs, and give rise to mature oligodendrocytes and astrocytes in vitro and in vivo. Notably, transplanted iOPCs contribute to functional recovery in a spinal cord injury (SCI) model without tumor formation. This study provides a simple strategy to generate functional self-renewing iOPCs and yields insights for the in-depth study of demyelination and regenerative medicine.

Attenuation of PRRX2 and HEY2 enables efficient conversion of adult human skin fibroblasts to neurons.

The direct conversion of accessible cells such as human fibroblasts to inaccessible cells, particularly neurons, opens up many opportunities for using the human model system to study diseases and discover therapies. Previous studies have indicated that the neuronal conversion of adult human skin fibroblasts is much harder than that for human lung fibroblasts, which are used in many experiments. Here we formally report this differential plasticity of human skin versus lung fibroblasts in their transdifferentiation to induced neurons. Using RNAseq of isogenic and non-isogenic pairs of human skin and lung fibroblasts at different days in their conversion to neurons, we found that several master regulators (TWIST1, TWIST2, PRRX1 and PRRX2) in the fibroblast Gene Regulatory Network were significantly downregulated in lung fibroblasts, but not in skin fibroblasts. By knocking down each of these genes and other genes that suppress the neural fate, such as REST, HES1 and HEY2, we found that the combined attenuation of HEY2 and PRRX2 significantly enhanced the transdifferentiation of human skin fibroblasts induced by ASCL1 and p53 shRNA. The new method, which overexpressed ASCL1 and knocked down p53, HEY2 and PRRX2 (ApH2P2), enabled the efficient transdifferentiation of adult human skin fibroblasts to MAP2+ neurons in 14 days. It would be useful for a variety of applications that require the efficient and speedy derivation of patient-specific neurons from skin fibroblasts.