犬モデルにおける ex vivo および in vivo 遺伝子治療のための代替遺伝子導入技術の開発

Introduction: Gene therapy have recently attracted much attention as a curative therapeutic option for inherited single gene disorders such as hemophilia. Hemophilia is a hereditary bleeding disorder caused by the deficiency of clotting activity of factor VIII (FVIII) or factor IX (FIX), and gene therapy for hemophilia using viral vector have been vigorously investigated worldwide. Toward further advancement of gene therapy for hemophilia, we have previously developed and validated the efficacy of novel two types of gene transfer technologies using a mouse model of hemophilia A. Here we investigated the efficacy and safety of the technologies in canine model. Especially, validations of technical procedures of the gene transfers for dogs were focused. Methods: Green fluorescence protein (GFP) gene were transduced into normal beagle dogs by ex vivo and in vivo gene transfer techniques. For ex vivo gene transfer, blood outgrowth endothelial cells (BOECs) derived from peripheral blood of normal dogs were transduced with GFP gene using lentivirus vector, propagated, fabricated as cell sheets, then implanted onto the omentum of the same dogs. For in vivo gene transfer, normal dogs were subjected to GFP gene transduction with non-viral piggyBac vector by liver-targeted hydrodynamic injections. Results: No major adverse events were observed during the gene transfers in both gene transfer systems. As for ex vivo gene transfer, histological findings from the omental biopsy performed 4 weeks after implantation revealed the tube formation by implanted GFP-positive BOECs in the sub-adipose tissue layer without any inflammatory findings, and the detected GFP signals were maintained over 6 months. Regarding in vivo gene transfer, analyses of liver biopsy samples revealed more than 90% of liver cells were positive for GFP signals in the injected liver lobes 1 week after gene transfers, then the signals gradually declined overtime. Conclusions: Two types of gene transfer techniques were successfully applied to a canine model, and the transduced gene expressions persisted for a long term. Toward clinical application for hemophilia patients, practical assessments of therapeutic efficacy of these techniques will need to be performed using a dog model of hemophilia and FVIII (or FIX) gene.博士（医学）・乙第1517号・令和3年12月21日© 2021, The Japanese Society for Regenerative Medicine. Production and hosting by Elsevier B.V. This is an open access article under the CC BY-NC-ND license (http://creativecommons.org/licenses/by-nc-nd/ 4.0/)

Recommendations related to the analytical equivalence assessment of gene panel testing

Advances in cancer genome care over the past few years have included the development of gene panel testing for various biomarkers. This article summarizes issues and provides recommendations related to analytical performance evaluations for new oncology gene panels. The scope of these recommendations includes comprehensive genomic profiling assays related to gene panel testing that uses histological or serum specimens to detect gene mutations. As a research project of the Japan Agency for Medical Research and Development Research on Regulatory Science of Pharmaceuticals and Medical Devices, we convened the working group committee that consisted of more than 30 experts from academia, industry, and government. We have discussed the points that should be considered to allow maximal simplification of assessments using clinical specimens in evaluating accuracy and limit of detection in equivalence and analytical performance for three years. We provide recommendations specific to each type of gene mutation as well as to reference standards or specimens used for evaluations. In addition, in order to facilitate the discussion on the analytical performance of gene panel tests by multidisciplinary tumor boards of hospitals, the present recommendations also describe the items that companies are expected to provide information on in their packaging inserts and reports, and the items that are expected to be discussed by multidisciplinary tumor boards. Our working group document will be important for participants in multidisciplinary tumor boards including medical oncologists and genome scientists, and developers of gene panels not only in Japan but also in other countries

Mineralization of hydroxyapatite upon a unique xanthan gum hydrogel by an alternate soaking process

We previously reported a xanthan gum (Xan) hydrogel showing excellent mechanical properties. Mineralization of hydroxyapatite (Hap) upon the Xan hydrogel would provide a unique biomaterial applicable for bone tissue engineering. Here, we show the mineralization of Hap upon the Xan hydrogel by means of an alternate soaking process. Hap was gradually grown upon the Xan-matrix surface with increasing number of soaking cycles due to the ionic interactions between calcium cations and carboxyl groups. Interestingly, the mineralization induced a microstructure change in the gel-matrix from a layered structure to a porous structure. The mechanical properties of the resulting Hap–Xan composite hydrogels were further investigated by a tensile test, where the Hap–Xan composite hydrogel with an appropriate amount of Hap (Xan/Hap = 2.7) was capable of approximately 370% elongation

Development of Cellphone-type Tele-operated Android Human Factors; Design

ABSTRACT This paper presents a newly developed portable human-like robotic avatar "Elfoid" which can be a novel communication medium in that a user can talk with another person in a remote location in such a way that they feel each other's presence. It is designed to convey individuals' presence using voice, human-like appearance, and touch. Thanks to its cellphone capability, it can be used at anytime, anywhere. The paper describes the design concept of Elfoid and argues research issues on this communication medium

How does telenoid affect the communication between children in classroom setting?

It needs to be investigated how humanoid robots may affect people in the real world when they are employed to express the presence, a feel of being there, in tele-communication. We brought Telenoid, a tele-operated humanoid robot, into a classroom at an elementary school to see how schoolchildren respond to it. Our study is exploratory and we focused on the social aspects that might facilitate communication between schoolchildren. We found that Telenoid affected the way children work as group. They participated in the group work more positively, became more spontaneous, and differentiated their roles. We observed that Telenoid's limited capability led them to change their attitudes so that they could work together. The result suggests that the limited functionality may facilitate cooperation among participants in classroom setting