On 12 August of this year, Professor Shigetaka Asano died. He was a celebrated Japanese hematologist and scientist; past director of the IMSUT (The Institute of Medical Science, The University of Tokyo) Hospital; an illustrious leader of the Japanese immunology, genetics and gene therapy communities; founder of the Japanese Society of Gene Therapy; recipient of many honors, including the Bälz Prize, the Okuma Prize from the Waseda University, and the Order of the Sacred Treasure with Gold Rays and Rosette (Figure 1); a joyous friend and colleague; and a lover of life. Those who had the privilege of working with him and studying and learning from him during his training and long academic career – sadly, I did not have that good fortune – know of his many scientific and academic accomplishments in immunology, genetics and medicine, including his molecular cloning of granulocyte-colony stimulating factor (G-CSF) and the development of recombinant human native G-CSF, the creation of the Japanese marrow donor program and the Japanese Public Cord Blood Banking system, and the uniting of the Japanese Societies of Hematology and Clinical Hematology. Those accomplishments have been cited more knowledgably in other commentaries by Professors Ozawa, Kodo and Gale1 who worked closely with Dr Asano. However, I did have the good fortune of getting to know him and of experiencing his joy during those early formative and conceptual years of gene therapy through visits to his office at the IMSUT Hospital and during the accompanying wonderful dinners that were made even more memorable by the warm sake. Those were parts of the experiences that helped to define the early concepts of gene therapy and that cemented my close relationship with the extraordinary Japanese gene therapy community. Over the years, Dr Asano and I were able to maintain contact with each other mostly through e-mails and seasonal messages. In one recent message, he sent me a copy of a painting that he had done (Figure 2). I was previously unaware of his devotion to art and to painting, although I have now come to understand that painting with colleagues was one of his human pleasures that brought them closer together. The painting is of a dark, gloomy and menacing tree trunk with a sprig of beautiful bright white flowers and a promising green shoot emerging from the trunk. The painting struck me as an expression of Professor Asano's appreciation of hope, truth and beauty, just as genetics, medical science and gene therapy promise relief from disease, as if to deliver on the promise of his scientific work. Sadly, Dr Asano's death underscores the wisdom of the great American poet Robert Frost, who reminds us of the transience of beauty. True as that might be, the joy and optimism of Dr Asano's life and the successes of his medical accomplishments remain with us, not in person, but as a legacy of his life and his accomplishments. Frost wrote, “Nothing gold can stay”. The gold of Asano is gone, but the gold of his legacy stays with us.

Human enhancement technologies are opening tremendous opportunities but also challenges to the core of what it means to be human. We argue that the goal of human enhancement should be to enhance quality of life and well-being not only of individuals but also of the communities they inhabit.

profound an effect as bariatric surgery, Lingvay said, suggesting that the hormone is just one ingredient among many. “It turns out that there are multiple gut hormones and neuroresponse phenomena that follow a gastric bypass— some of them affect the satiety signals, some affect islet production of insulin,” Wolfe said. What that translates to is that “[t]here’s no single potion that’s going to solve this problem.” It’s looking increasingly likely that the potion—a fix for obesity and type 2 diabetes that doesn’t require major surgery— will take the form of combination drugs targeting more than 1 pathway. Berthoud and other researchers are beginning to study the combined antidiabetic and body weight–lowering effects of GLP-1 with other hormones, including other insulinstimulating hormones. Some patients, such as those with lower body weights, may get results from a treatment protocol that combines pharmaceutical therapy with a nonsurgical procedure that reproduces the effects of gastric bypass, Kashyap said. Less-invasive endoscopic approaches currently in development that target the foregut, where a key surgery-specif ic mechanism may reside, include the placement of a tubeshaped liner and mucosal resurfacing with thermal ablation. “Bariatric surgery is one extreme form of treating diabetes,” Kashyap said, “but 99% of patients would like to improve their health and diabetes without having the surgery.” For Lingvay, the less-invasive solution comes back to the role of the brain: “If we can figure [out] what’s that effect on the brain that keeps patients from eating, we would have an answer to how to do this without surgery.”

Here we propose new module-based approaches to identify differentially regulated network sub-modules combining temporal trajectories of expression profiles with static network skeletons. Starting from modules identified by network clustering of static networks, our analysis refines pre-defined genesets by partitioning them into smaller homogeneous sets by non-paramettric Bayesian methods. Especially for case-control time series data we developed multi-time point discriminative models and identified each network module as a mixture or admixture of dynamic discriminative functions. Our results shows that our proposed approach outperformed existing geneset enrichment methods in simulation studies. Moreover we applied the methods to neural stem cell differentiation data, and discovered novel modules differentially perturbed in different developmental stages.

Despite being prohibited by the World Anti-Doping Agency (WADA), blood manipulations such as the use of recombinant human erythropoietin and blood transfusions are a well-known method used by athletes to enhance performance. Direct detection of illicit blood manipulation has been partially successful due to the short detection window of the substances/methods, sample collection timing, and the use of sophisticated masking strategies. In response, WADA introduced the athlete biological passport (ABP) in 2009, which is an individualised longitudinal monitoring approach that tests primarily haematologic biomarkers of doping in order to identify atypical variability in response(s) in athletes, highlighting a potential doping violation. Although the implementation of the ABP has been an encouraging step forward in the quest for clean/drug-free sport, this detection method has some limitations. To reduce the risk of being detected by the ABP method, athletes are now resorting to microdoses of prohibited blood boosting substances to prevent abnormal fluctuations in haematologic biomarkers, thereby reducing the sensitivity of the ABP detection method. Recent studies from numerous laboratories, including our own, have confirmed the potential of transcriptomic microarrays, which can reveal distinct changes in gene expression after blood manipulations, to enhance the ABP. There is, therefore, an urgent need to intensify research efforts that involve transcriptomics and other state-of-the-art molecular methods, collectively known as “omics”, e.g., proteomics (proteins) and metabolomics (metabolites), in order to identify new and even more robust molecular signatures of blood manipulation that can be used in combination with the ABP and, intriguingly, even as a stand-alone test.

An experimental approach for gene therapy of spinomuscular atrophy has been reported to prevent development of the neuromuscular features of this lethal and previously untreatable disorder. The approach involves treatment of patients suffering from SMN1-associated infantile form of the disease with a splice-switching antisense oligonucleotide (ASO) that corrects aberrant splicing of the nearly identical SMN2 gene to allow the generation of functional SMN protein, thereby mitigating the development of the disease. This technique represents the first apparently effective therapy for spinal muscular atrophy (SMA) and an important documentation for ASO technology for therapy of neurodegenerative disease. These results with one form of SMA are likely to be relevant for similar applications to other SMA types and are likely to inspire application to a number of other intractable neurodegenerative diseases such as Huntington’s disease, amyotrophic lateral sclerosis and possibly even the extremely common Parkinson’s and Alzheimer’s diseases and others. Nevertheless, the scientific and medical importance of this advance is marred by a pricing policy by the corporate sponsors that may complicate accessibility of the drug for some desperate patients.

Introduction and background Doping is an ancient and familiar part of Sport. For the first two decades of the twentieth century, the tug of war was an established and formal competitive event of the early re-established modern summer Olympic games. Like most athletic events, the goal and the inevitable outcome was victory for one side or another. Although the tug of war event itself has vanished from the Olympic Games and from elite international Sport, it has come to be replaced by an analogous but far more enduring form of tug of war: i.e., between doping and anti-doping forces in Sport. And, as recent history has suggested, this tug of war tends to conclude not in definitive victories but rather in exchanges of advances as forms of doping and cheating come to be detected and then replaced by more sophisticated and technologically updated techniques that in turn lead to anti-doping technical advances, and so on, ad infinitum.

The armamentarium of modern genetics and gene therapy has recently expanded with the rapid development of the tools of “genome editing,” including zinc-finger nucleases, TALENs, and CRISPR/Cas9 techniques that allow exquisitely targeted sequence modifications. An alluring application of such technology is somatic cell genome editing for the prevention and therapy of genetic disease. Most scientific, ethics, and policy agencies support somatic cell applications of genome editing; the gene therapy community has understandably taken deep interest because we represent the major biomedical conduit through which the technology will eventually be translated to clinical application. But it takes little imagination to recognize that there are other more contentious potential applications of genome editing, especially those related to the potential for germ-line modification. The recent publication of an article from China reporting an attempt to edit the human hemoglobin locus using CRISPR/Cas9 in early-stage human embryos added urgency to these concerns.1Liang P Xu Y Zhang X Ding C Huang R Zhang Z et al.CRISPR/Cas9-mediated gene editing in human tripronuclear zygotes Protein Cell. 2015; 6: 363-372Google Scholar In response, the American Society for Gene and Cell Therapy (ASGCT) and the Japanese Society of Gene Therapy (JSGT) issued a joint position statement last year supporting rigorous basic and clinical research to perfect safe and effective clinical applications of genome editing at the somatic cell level.2Friedmann T Jonlin EC King NM Torbett BE Wivel NA Kaneda Y ASGCT and JSGT joint position statement on human genomic editing Mol Ther. 2015; 23: 1282Abstract Full Text Full Text PDF Scopus (41) Google Scholar However, the statement expressed concern that genome editing in human gametes, zygotes, and embryos carries serious scientific, ethical, and societal implications. The statement noted that there is no broad scientific and societal consensus about these issues and that the effects of inherited genomic modification cannot readily be evaluated scientifically or ethically over the long time course of a reproductive human application. The poorly understood long-term effects of inherited human genetic modification could encourage unwise and ill-prepared manipulation of human genetic evolution, lead toward the highly contentious and fraught area of human genetic enhancement of normal human traits, and provide tacit rationalization of an illusory neo-eugenics movement. The Societies therefore recommended that clinical genome editing applications with reproductive potential should be disallowed, at least until a scientific and international societal consensus was reached regarding research on these approaches. An important next step occurred with the holding of a public forum at the US National Academy of Sciences 1–3 December 2015, the International Summit on Human Gene Editing, chaired by David Baltimore and sponsored by the US National Academy of Sciences, the US Academy of Medicine, the Chinese Academy of Sciences, and the Royal Society of London. The goal of the summit was to discuss the scientific, ethical, and governance issues associated with human gene editing research in an open and inclusive forum, and to develop scientific and policy recommendations on the means to move forward. Participants in the meeting included many of the scientists responsible for discovering and developing genome editing technology, ethicists, policy makers from around the globe, historians, physicians, disease and disability interest groups, and both the lay and scientific press. The sessions included detailed presentation and discussions of the scientific background and basic research aspects of gene editing technologies; the application of gene editing technology and human germ-line modification; societal implications of emerging technologies; the potential for application to somatic cell therapy; comparison of national US and international governance, regulation and control policies; and potential dilemmas related to equity. ASGCT member and leading gene therapy physician–scientist Adrian Thrasher served on the planning committee for the summit, representing the British Royal Society. Thrasher moderated a very important session on somatic gene editing, at which several ASGCT members spoke, including Matthew Porteus and Fydor Urnov. They and many other presenters eloquently explained that basic and clinical research on the use of somatic gene editing to treat human disease was already progressing, involved no new ethical or regulatory concerns, and should not be confused with germ-line gene editing. Other speakers contended that germ-line genetic modification by genome editing of gametes, zygotes, or embryos would not fill a compelling clinical need that could not be more efficiently filled by the currently available technology of pre-implantation genetic diagnosis and even by the rapidly increasing effectiveness of somatic cell gene therapy. That leaves only those rare cases involving two potential parents who are both homozygous for a genetic disorder and whose offspring would all be similarly affected. Admittedly, germ-line modification by genome editing might indeed offer a reproductive opportunity for such people, although those situations would be exceedingly rare. Another theme was the very disparate international philosophies, regulations, and funding of assisted reproductive technologies, which would be a component of any germ-line editing approach. Finally, there were extensive discussions on how or whether lines can be drawn between correction vs. “enhancement” of the germ line, how any such technologies could be applied equitably across the globe or even within technologically developed societies, and ways to reconcile germ-line editing with evolving concepts of disability. All slides and discussions from the general sessions of the summit can be viewed at http://www.nationalacademies.org/gene-editing/Gene-Edit-Summit/index.htm. Based on these presentations and discussions, the summit's planning committee formulated a series of recommendations for the development and human applications of genome editing.3The National Academies of Sciences, Engineering, and Medicine, 2015 On Human Gene Editing: International Summit Statement Press release (3 December 2015)Google Scholar Next steps include review by the parent sponsoring societies and planned follow-up meetings to be sponsored by the National Academies. We summarize the central recommendations below. •Basic and preclinical research. Research on gene editing in both somatic tissues and human embryos or gametes was strongly supported, subject to appropriate legal and ethical oversight. The statement, however, warned: “If, in the process of research, early human embryos or germline cells undergo gene editing, the modified cells should not be used to establish a pregnancy.”•Clinical use: somatic. This section of the statement specifically stressed that valuable clinical applications in somatic cells are ongoing and should continue, singling out examples including treating sickle cell anemia or cancer via editing in blood cells. The statement indicated that research on the risks and benefits of somatic gene editing approaches should continue and that the existing regulatory framework developed for gene therapy is sufficient to encompass these approaches.•Clinical use: germ line. This section of the statement summarized the current scientific, practical, and ethical objections to human germ-line editing, including risks of passing on off-target effects, mosaicism, population risks from the introduction of genetic changes, considerations regarding implications for both the individual being treated and future generations, the impossibility of removing alterations once introduced, the possibilities that permanent genetic “enhancements” could exacerbate inequities or be used coercively, and the moral and ethical implications of purposefully altering human evolution. This central section of the statement concluded: “It would be irresponsible to proceed with any clinical use of germline editing unless and until (i) the relevant safety and efficacy issues have been resolved, based on appropriate understanding and balancing of risks, potential benefits, and alternatives, and (ii) there is broad societal consensus about the appropriateness of the proposed application. Moreover, any clinical use should proceed only under appropriate regulatory oversight. At present, these criteria have not been met for any proposed clinical use.…However, as scientific knowledge advances and societal views evolve, the clinical use of germline editing should be revisited on a regular basis.”•Future steps. The statement called for continued international discussion and cooperation to establish norms and harmonize potential acceptable uses of human germ-line editing, recommending that the sponsoring academies lead ongoing forums on these topics, continuing to bring together all nations and perspectives to address the evolving issues. The Genome Editing Summit has provided us with a prudent and cautious high bar that strives to protect legitimate and universally desired tools for amelioration of suffering while recognizing the deep ethical quandaries that accompany this new potential capability of shaping aspects of our human inheritance. Gratifyingly, these recommendations echo most of the recommendations proposed by the previous study of inherited genome modification carried out by the American Association for the Advancement of Science4Frankel MS Chapman AR Designing Our Descendants: The Promises and Perils of Genetic Modifications. Johns Hopkins Press, Baltimore, MD2003Google Scholar and the joint ASGCT/JSGT policy statement on the use of genome editing.2Friedmann T Jonlin EC King NM Torbett BE Wivel NA Kaneda Y ASGCT and JSGT joint position statement on human genomic editing Mol Ther. 2015; 23: 1282Abstract Full Text Full Text PDF Scopus (41) Google Scholar This is only the beginning of a long and complex process that will be influenced differently in different jurisdictions with disparate social, jurisprudential, and religious traditions. There will probably be no uniform final consensus, but the scientific and public discussion will point the way toward fruitful and beneficial uses of this powerful new technology. Interested parties and ASGCT members should plan to attend a special workshop on the topic of genome editing that will cover many of these issues and include presentations from leaders in the relevant scientific, ethical, and regulatory fields, to be held on 4 May 2016 as part of the 2016 ASGCT annual meeting in Washington, DC.

The American Society for Gene and Cell Therapy (ASGCT) and the Japan Society of Gene Therapy (JSGT) (collectively, "Our Societies") recognize the great scientific advancement represented by the techniques of genome editing and their vast potential value for an improved understanding and possible treatment of human disease. These techniques provide uniquely powerful tools for generating models of human disease, for characterizing the molecular and biochemical basis for pathogenesis, and for suggesting approaches to definitive correction of genetic defects underlying much of human disease. However, our Societies also recognize that the application of genome editing under some circumstances poses very serious ethical problems for which there is no scientific or general societal consensus and that should be considered as inappropriate human genetic manipulation until and unless serious scientific and ethical concerns can be resolved. Somatic cell genome editing Gene editing methods will make great contributions to the study, understanding, and treatment of human disease. At the somatic cell level, certain types of genome editing will have legitimate scientific and medical applications because they have potential advantages over less precise gene transfer technologies. Our Societies consider current scientific methodology to be sufficient to clarify and correct the inevitable issues related to safety and efficacy of somatic cell gene editing. Although the ethical concerns arising from somatic cell gene editing naturally merit broad scientific and societal discussion, our Societies consider it unlikely that somatic cell gene editing will give rise to new or unique ethical concerns substantially different from those associated with other forms of research and therapy that have already been well discussed. Embryonic cell genome editing and germ-line modification Our Societies recognize that gene editing in human embryonic cells or in stem cells destined for use in creation of a human being may in the long run have the potential for correcting genetic defects associated with genetic disease, not only in patients, but theoretically in future generations as well. Our Societies also recognize that this area of research is complicated by a number of major and unresolved ethical concerns associated with germ-line genetic modification. Because research subjects would include not only embryos but also future generations, the difficulties of long-term follow-up raise ethical, practical, and scientific hurdles. The requirement that the results of an experiment be susceptible to analysis and characterization before further applications are undertaken cannot be met with human germ-line modification with current methods, because the results of any such manipulation could not be analyzed or understood for decades or generations—a situation incompatible with ethical imperatives and with the scientific method. Finally, our Societies consider it essential to develop effective social and policy mechanisms for carrying out broad and deep discussions of human germ-line alteration so as effectively to understand and balance the individual, familial, societal, and perhaps even species-level rights, needs, interests, and values affected by this rapidly advancing science. In addition, our Societies consider the currently available genome editing technologies to be inadequately understood and developed for carrying out gene editing or other forms of genetic manipulation of human reproductive cells, pluripotent stem cells, or embryonic tissues with the potential for incorporation into viable human zygotes for either disease prevention or enhancement. Acceptable technology would require a far greater understanding and control of gene interactions than is currently available, elimination of off-target effects, and absolute prevention of mosaicism in resulting human embryos or zygotes. Our Societies consider that such applications with the current technology would not be adequately controlled and would therefore be ethically unacceptable. In the meantime, research on gene editing in reproductive cells in various animal model systems, including primates, should be continued. This has the potential to clarify technical problems associated with gene editing and to devise solutions that will strengthen the technology. Such efforts could also lay the groundwork for establishing guidelines for eventual studies with nonviable human embryos. Summary Our Societies consider these safety and ethical concerns to be sufficiently serious to support a strong stance against gene editing in, or gene modification of, human cells to generate viable human zygotes with heritable germ-line modifications. Even with technical advances that may eventually solve the safety and mosaicism problems, our Societies conclude that there are not ethically acceptable ways to conduct embryonic gene editing or other germ-line modifications because the results of such experiments are not susceptible to long-term evaluation in a scientifically reasonable time scale. For these reasons, our Societies support a strong ban on human germ-line gene editing or other germ-line genetic modification unless and until these technical and ethical problems can be solved, broadly and deeply discussed, and societal consensus reached.

The field of gene therapy in general has made great progress during the past decade and is now delivering real treatment to patients with a number of difficult and previously intractable diseases. The central nervous system (CNS) has always represented a tempting but difficult target for gene-based therapies, not only because of the immense complexity of CNS function but also because of the difficulty in identifying those genes or pathways whose dysregulation is truly causative in CNS disease. Fortunately, and perhaps a little surprisingly, neurodevelopmental and neurodegenerative diseases are increasingly becoming amenable to genetic therapy. The past few years have seen the prevention and the amelioration of a number of CNS disorders, and the pace of advances in neuroscience and gene transfer technology certainly points to startling advances soon to come. It is comforting finally to be on the verge of having truly effective therapy for more and more of these disorders and to realize that the CNS has provided incontrovertible proof of principle for gene therapy.

Transcriptomic studies of murine D3 embryonic stem (ES) cells deficient in the purinergic biosynthetic function hypoxanthine guanine phosphoribosyltransferase (HPRT) and undergoing dopaminergic neuronal differentiation has demonstrated a marked shift from neuronal to glial gene expression and aberrant expression of multiple genes also known to be aberrantly expressed in Alzheimer's and other CNS disorders. Such genetic dysregulations may indicate some shared pathogenic metabolic mechanisms in diverse CNS diseases.

The mechanisms by which mutations of the purinergic housekeeping gene hypoxanthine guanine phosphoribosyltransferase (HPRT) cause the severe neurodevelopmental Lesch Nyhan Disease (LND) are poorly understood. The best recognized neural consequences of HPRT deficiency are defective basal ganglia expression of the neurotransmitter dopamine (DA) and aberrant DA neuronal function. We have reported that HPRT deficiency leads to dysregulated expression of multiple DA-related developmental functions and cellular signaling defects in a variety of HPRT-deficient cells, including human induced pluripotent stem (iPS) cells. We now describe results of gene expression studies during neuronal differentiation of HPRT-deficient murine ESD3 embryonic stem cells and report that HPRT knockdown causes a marked switch from neuronal to glial gene expression and dysregulates expression of Sox2 and its regulator, genes vital for stem cell pluripotency and for the neuronal/glial cell fate decision. In addition, HPRT deficiency dysregulates many cellular functions controlling cell cycle and proliferation mechanisms, RNA metabolism, DNA replication and repair, replication stress, lysosome function, membrane trafficking, signaling pathway for platelet activation (SPPA) multiple neurotransmission systems and sphingolipid, sulfur and glycan metabolism. We propose that the neural aberrations of HPRT deficiency result from combinatorial effects of these multi-system metabolic errors. Since some of these aberrations are also found in forms of Alzheimer's and Huntington's disease, we predict that some of these systems defects play similar neuropathogenic roles in diverse neurodevelopmental and neurodegenerative diseases in common and may therefore provide new experimental opportunities for clarifying pathogenesis and for devising new potential therapeutic targets in developmental and genetic disease.

Mutations in the gene encoding the purine biosynthetic enzyme hypoxanthine-guanine phosphoribosyltransferase (HPRT) cause the intractable neurodevelopmental Lesch-Nyhan disease (LND) associated with aberrant development of brain dopamine pathways. In the current study, we have identified an increased expression of the microRNA miR181a in HPRT-deficient human dopaminergic SH-SY5Y neuroblastoma cells. Among the genes potentially regulated by miR181a are several known to be required for neural development, including Engrailed1 (En1), Engrailed2 (En2), Lmx1a and Brn2. We demonstrate that these genes are down-regulated in HPRT-deficient SH-SY5Y cells and that over-expression of miR181a significantly reduces endogenous expression of these genes and inhibits translation of luciferase plasmids bearing the En1/2 or Lmx1a 3'UTR miRNA-binding elements. Conversely, inhibition of miR181a increases the expression of these genes and enhances translation of luciferase constructs bearing the En1/2 and Lmx1a 3'UTR miRNA-binding sequences. We also demonstrate that key neurodevelopmental genes (e.g. Nurr1, Pitx3, Wnt1 and Mash1) known to be functional partners of Lmx1a and Brn2 are also markedly down-regulated in SH-SY5Y cells over-expressing miR181a and in HPRT-deficient cells. Our findings in SH-SY5Y cells demonstrate that HPRT deficiency is accompanied by dysregulation of some of the important pathways that regulate the development of dopaminergic neurons and dopamine pathways and that this defect is associated with and possibly due at least partly to aberrant expression of miR181a. Because aberrant expression of miR181a is not as apparent in HPRT-deficient LND fibroblasts, the relevance of the SH-SY5Y neuroblastoma cells to human disease remains to be proven. Nevertheless, we propose that these pleiotropic neurodevelopment effects of miR181a may play a role in the pathogenesis of LND.