Histopathological characteristics of human non-tumor thyroid tissues in a long-term model of adenomatous goiter xenografts in the NOD/Shi-scid,IL-2Rγnull mouse

There is a growing need for modeling the human thyroid to link data obtained from animals to humans because of its sensitivity to radiation exposure and endocrine disruption chemicals. In a scid mouse model produced by transplanting human thyroid tissues, leakiness and thymic lymphoma that occurs spontaneously in the scid mouse can complicate the interpretation of experimental results. Considering that the NOD.Cg-Prkdc^scid Il2rg^tm1Sug/Jic mouse (NOD/Shi-scid, IL-2Rγ^null or NOG mouse) may be a better host because this strain has low incidence of leakiness and thymic lymphoma, we have evaluated the potential of a model that allows long-term observation of non-tumor human thyroid tissues in this mouse. We transplanted tissues of human adenomatous goiter into NOG mice and examined the tissues histopathologically. The morphology of human adenomatous goiter tissues was maintained from 24 to 44 weeks after transplantation in NOG mice with no noted differences between donor-matched tissues or the weeks after transplantation. The tissues expressed thyroglobulin protein and mRNA as well as thyroperoxidase. Endothelial cells originating from human were found in the transplanted tissues and were thought to be a characteristic of this model. The intactness of the tissues before transplantation was found to affect the rate of tissue engraftment. From the present results we have concluded that transplanted thyroid tissues in NOG mice maintain the histopathological characteristics of their origin for long terms. Therefore this model was thought feasible for toxicity evaluation.     

人牙髓间充质干细胞致瘤性试验

目的 观察人牙髓间充质干细胞（HDP-MSCs）对免疫缺陷小鼠的致瘤性作用。方法 雌性NOG小鼠随机分为4组：对照组和HDP-MSCs低、中、高剂量（2×10⁶、5×10⁶和2×10⁷ cells/kg）组,每组20只;双后肢交替im给药,每周给药1次,共给药6个月（27次）;试验期间进行大体观察、体质量检查和肿瘤触诊,给药结束进行解剖检查、脏器检查和组织病理学检查。结果 对照组死亡2只动物,其中1只动物颈部出现肿块;HDP-MSCs低剂量死亡1只动物,另有1只动物出现弓背;中剂量组1只动物濒死;高剂量动物未见明显异常;低、中、高剂量组触诊未发现肿瘤。小鼠体质量无异常。解剖后对照组和HDP-MSCs低、中剂量个别动物发现有肿块。各剂量组脏器质量及系数无异常、组织病理学检查无药物相关性病变。结论 HDP-MSCs在本试验剂量下连续im给药6个月,致瘤试验结果为阴性。     

Establishment of Long‐Term Model throughout Regular Menstrual Cycles in Immunodeficient Mice

Citation Kikuchi‐Arai M, Murakami T, Utsunomiya H, Akahira J, Suzuki‐Kakisaka H, Terada Y, Tachibana M, Hayasaka S, Ugajin T, Yaegashi N. Establishment of long‐term model throughout regular menstrual cycles in immunodeficient mice. Am J Reprod Immunol 2010 Problem The number of uterus natural killer (NK) cells change through the menstrual cycle, but the origin of uterus NK cell was not unclear. Our aims are to study whether we can reproduce repetition of menstrual cycle and to reveal the origin of uterus NK cells. Method of study Endometrial samples were obtained from fertile women, and the tissues were transplanted into ovariectomized non‐obese diabetic (NOD)/severe combined immunodeficiency (SCID)/γCnull (NOG) mice. Mice were treated with sex hormones which were in accord with human menstrual cycle. Results The replants showed similar histological changes as in eutopic endometrium repeatedly. CD56‐positive, CD16‐negative NK cells increased significantly during the treatment with estradiol and progesterone combination. Conclusion Histological assessment demonstrated that this model of NOG mice repeatedly exhibited regular menstrual cycles, and this model mimicked not ‘ectopic endometrium’, but ‘eutopic endometrium’ in humans. Change in number of NK cells suggested that NK cell might be derived from the endometrium.     

Development of thrombopoietin receptor agonists for clinical use

Summary.  Thrombopoietin (TPO) is an essential hematopoietic cytokine for megakaryopoiesis. In 2002, we demonstrated that pegylated-recombinant human megakaryocyte growth and development factor (PEG-rHuMGDF) increased platelet counts in patients with chronic immune thrombocytopenic purpura (ITP) in a Phase I/II clinical trial. After the cessation of clinical trials of PEG-rHuMGDF because of severe thrombocytopenia or pancytopenia due to the development of the neutralizing antibody cross-reacting with endogenous TPO, second generation non-immunogenic TPO receptor agonists have been developed. A small molecule eltrombopag and Romiplostim were approved for clinical use by FDA in 2008 to treat patients with chronic ITP who are refractory to the prior therapy. Although the efficacy of both TPO receptor agonists is convincing for the refractory ITP, further investigation is necessary to assess the potential long-term side effects and clinical applications of these therapies for other thrombocytopenic conditions.     

Evaluation of domain of unknown function 1220 (DUF1220) for detection of human genome by quantitative polymerase chain reaction: Potential use in assessing the biodistribution of transplanted therapeutic human cells

The biodistribution profile of cell-based therapy products in animal models is important for evaluation of their safety and efficacy. Because of its quantitative nature and sensitivity, the quantitative polymerase chain reaction (qPCR) is a useful method for detecting and quantifying xenogeneic cell-derived DNA in animal models, thereby allowing a biodistribution profile to be established. Although the restriction endonuclease family from Arthrobacter luteus (Alu) of repetitive elements in human genome sequences has been used to assess the biodistribution of human cells, high background signals are detected. In the present study, we evaluate the potential of domain of unknown function 1220 (DUF1220), which is a human lineage-specific, multiple-copy gene similar to Alu sequences, for such analysis. Using qPCR analysis for DUF1220, human genome could be detected against a mouse genome background at a level comparable to that of Alu sequences with no detectable background signals. Moreover, using this approach, the human genome could be distinguished from the cynomolgus monkey genome. Further investigation of the quantitative aspects of this DUF1220-based qPCR assay might prove its usefulness for biodistribution studies of human cells transplanted into animals in the future.     

骨形成蛋白4基因与NOG基因的交互作用与非综合征性唇腭裂的关系

目的:探讨骨形成蛋白4基因538T/C(BMP4 538T/C)变异与NOG基因615G/C(NOG 615G/C)变异之间交互作用在非综合征性唇腭裂(nsCL/P)发生中的意义。方法:从两家综合性医院中选择200名nsCL/P患者和200名对照,两组在年龄、性别、家庭社会经济状况方面相匹配。应用聚合酶链反应-限制性片段长度多态性(PCR-RFLP)方法检测BMP4538T/C、NOG 615G/C的多态性;运用相加模型分析BMP4 538T/C与NOG 615G/C变异之间的交互作用。结果:用非条件Logis-tic回归模型调整父母亲怀孕时的年龄和文化程度后,BMP4 538T/C与NOG 615G/C变异之间对nsCL/P患病具有正交互作用,归因交互效应2.05,归因交互效应百分比46.28%,纯因子间归因交互效应百分比59.77%,交互效应指数2.49。结论:BMP4538T/C与NOG 615G/C变异之间交互作用在该研究人群nsCL/P患病中具有重要意义。     

一个中国近端指(趾)骨间关节黏连家系NOG基因新突变鉴定

目的 检测一个中国近端指(趾)骨间关节黏连家系的致病基因突变.方法 收集该家系患者和家系成员的临床资料,采集外周血提取基因组DNA.运用聚合酶链式反应和Sanger测序法筛查先证者的NOG和GDF5基因.确定突变位点后,在家系成员中进行共分离分析并在200名正常对照中筛查该突变.结果 该家系患者中存在NOG基因c.502 T＞C错义突变,该突变导致其编码蛋白Noggin第168位氨基酸由苯丙氨酸变为亮氨酸(p.F168L).在家系正常成员和200名正常对照中未检测到相同突变.该突变在dbSNP、ExAC等数据库中未见报道.在GDF5基因上未发现可疑突变.结论 NOG基因c.502 T＞C错义突变是该家系发病的原因.     

Multiple synostoses syndrome: Clinical report and retrospective analysis

Multiple synostoses syndrome (SYNS1; OMIM# 186500) is a rare autosomal dominant disorder reported in a few cases worldwide. We report a Chinese pedigree characterized by proximal symphalangism, conductive hearing loss, and distinctive facies. We examined the genetic cause and reviewed the literature to discuss the pathogeny, treatment, and prevention of SYNS1. Audiological, ophthalmological, and radiological examinations were evaluated. Whole‐exome sequencing (WES) was performed to identify mutations in the proband and her parents. Sanger sequencing was used to verify the results for the proband, parents, and grandmother. The literature on the genotype–phenotype correlation was reviewed. The patient was diagnosed with multiple synostoses syndrome clinically. WES and bioinformatic analysis revealed a novel missense mutation in the NOG gene, c.554C>G (p.Ser185Cys), cosegregated in this family. The literature review showed that the phenotype varies widely, but the typical facies, conductive hearing loss, and proximal symphalangism occurred frequently. All reported mutations are highly conserved in mammals based on conservation analysis, and there are regional hot spots for these mutations. However, no distinct genotype–phenotype correlations have been identified for mutations in NOG in different races. Regular systematic examinations and hearing aids are beneficial for this syndrome. However, the outcomes of otomicrosurgery are not encouraging owing to the regrowth of bone. This study expanded the mutation spectrum of NOG and is the first report of SYNS1 in a Chinese family. Genetic testing is recommended as part of the diagnosis of syndromic deafness. A clinical genetic evaluation is essential to guide prevention, such as preimplantation genetic diagnosis.