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Prenatal Treatment of Down Syndrome to Improve Brain Development and Neurocognition

唐氏综合症的产前治疗可改善大脑发育和神经认知

基本信息

批准号：
10267123
负责人：
Diana Bianchi
金额：
$ 118.62万
依托单位：
NATIONAL HUMAN GENOME RESEARCH INSTITUTE
依托单位国家：
美国
项目类别：
财政年份：
资助国家：
美国
起止时间：
至
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10267123
关键词：
Adult Affect Age Amniocentesis Aneuploidy Angiogenic Factor Animals Anti-Inflammatory Agents Antioxidants Apigenin Autopsy Brain Cell Cycle Cell Line Cessation of life Chorionic Villi Sampling Chromosome 21 Chromosomes Collaborations Collection Congenital Abnormality Congestive Cultured Cells DNA Repair Data Defect Development Diagnosis Diagnostic Procedure Diffusion Magnetic Resonance Imaging Discrimination Down Syndrome Embryo Exhibits Exploratory Behavior Extinction (Psychology)FDA approved Female Fetus Fibroblasts France Gene Expression Gene Expression Profiling Gene Proteins Genes Genetic Transcription Genome Genotype Goals Growth Hepatic Hippocampus (Brain)Histology Human IL1A gene IL7 gene In Vitro Individual Infant Inflammation Inflammatory Intellectual functioning disability Interleukin-10 Karyotype Laboratories Language Development Learning Life Liver Longevity Male Sterility Medical center Memory Modeling Molecular Motor Mus Necrosis Neurocognition Neuronal Differentiation Orthologous Gene Oxidative Stress Partner in relationship Pathologic Pathway interactions Pharmaceutical Preparations Phenotype Placenta Pre-Clinical Model Pregnant Women Principal Component Analysis Proteomics Publications Renal pelvis Reporting Rodent Safety Second Pregnancy Trimester Signal Transduction Testing Therapeutic Effect Toddler Training Translating Treatment Efficacy Trisomy VEGFA gene Ventricular Septal Defects Visual Weight Work antenatal autistic behavior test behavioral phenotyping brain abnormalities congenital anomaly cytokine defense response drug candidate efficacy testing experimental study fetal genome-wide hepatic necrosis improved in vivo induced pluripotent stem cell insight live cell imaging male malformation mouse Trisomy 16 mouse Ts65Dn mouse model myelination neonate nerve stem cell nestin protein neurogenesis neurotrophic factor novel postnatal prenatal prenatal therapy protein expression pup response routine screening safety testing sex single-cell RNA sequencing skills synaptogenesis therapeutic target touchscreen transcriptome transcriptomics treatment response

项目摘要

During the past year, we achieved the following objectives for each of our goals: 1) We generated a large collection of induced pluripotent stem cells (iPSCs) from individuals with trisomy 21 (T21) and age/sex matched euploid controls (Eup). The iPSCs were then differentiated into neural progenitor cells (NPCs). Similar to fibroblasts and iPSCs from individuals with T21, NPCs from individuals with T21 exhibited slower growth versus age and sex matched euploid lines. To get better insights into the molecular mechanisms underlying atypical brain development in fetuses with DS, we performed transcriptome analyses on fibroblasts, iPSCs, and NPCs. The results demonstrated dysregulation across multiple pathways including cell cycle, DNA damage/repair, inflammation, and oxidative stress. Principal component analysis revealed high levels of variability across NPC lines but not across matched iPSC lines. To better understand this variability within and between NPC lines, single-cell RNA sequencing was performed, and analysis is currently ongoing. In addition to transcriptomic approaches, ongoing proteomic studies of iPSC and NPC lines will help to identify translational changes that result from genome wide transcriptional dysregulation and further validate our transcriptome findings. Live cell imaging is being used to evaluate cell cycle defects, increased oxidative stress and inflammation in DS derived NPCs compared to euploid age and sex matched individuals. 2) We have previously reported major differences in the molecular, cellular and behavioral phenotypes of the Ts65Dn, Dp(16)1/Yey and Ts1Cje mouse models of DS (Aziz, Guedj et al, 2018). To identify the mouse model of DS that most closely mimics the human phenotype we are continuing our assessment of different mouse models across the lifespan. Prenatally, we are particularly focused on identifying embryonic and placental pathologic markers. Postnatally, we are using behavioral tests that are like those used in human infants, as well as diffusion tensor imaging and stereology to identify endpoints that can be used to test the efficacy of therapies. In one set of experiments, Dp(16)/1Yey and Ts1Cje males were mated to C57BL/6 females and Ts65Dn females to C57BL/6XC3Sn males. At day E18.5, dams were euthanized. Embryos and placentas were examined blindly for weight and size. A subset (34 euploid, 34 trisomic) of mice was examined for malformations. Euploid embryos had no congenital anomalies; one was demised. Hepatic necrosis was seen in 6/12 (50%) of Dp(16)1/Yey and 1/12 (8%) Ts1Cje embryos; hepatic congestion/inflammation was observed in 3/10 (30%) Ts65Dn embryos. Renal pelvis dilation was seen in 5/12 (42%) Dp(16)1/Yey, 5/10 (50%) Ts65Dn and 3/12 (25%) Ts1Cje embryos. One Ts65Dn and one Dp(16)1/Yey embryo had an aortic outflow abnormality. Two Ts1Cje embryos had ventricular septal defects. Ts65Dn placentas had increased spongiotrophoblast necrosis. The presence of liver abnormalities in all three mouse models of Down syndrome (10/34) is a novel finding. Renal pelvis dilation was also common (13/34). Differences in placental histology were also observed between strains. This work has been submitted for publication. Future research will examine autopsy material to determine if this finding is relevant to humans with Down syndrome. Studies from our laboratory have shown that Ts65Dn mice have the most significant brain abnormalities, but this model is not optimal because males are sterile and its genome contains triplication of non-orthologous genes on MMu17, including Arid1B, a gene that is involved in an autistic phenotype. In collaboration with the Herault Lab (IGBMC, France), we recently rederived a novel mouse model (Ts65Yah) that carries the same Mmu16 trisomy as the Ts65Dn without the non-orthologous Mmu17 trisomy. We analyzed gene expression changes in the Ts66Yah mouse model and confirmed the exclusive trisomy of Mmu16 orthologous genes without the presence of Mmu17 non-orthologous genes. E18.5 gene expression data has confirmed major differences between the four mouse models used in our studies and demonstrated that the Ts66Yah model is the one that most closely mimics the human karyotype. Infants with Down syndrome (DS) demonstrate delays in motor development and expressive language acquisition. We translated the CANTAB Visual Discrimination (VD) and Extinction tasks (used in human infants and toddlers with Down syndrome) to adult rodent touchscreen paradigms to investigate hippocampal learning and cortical inhibitory control in the Dp(16)1/Yey, Ts65Dn and Ts1Cje mouse models. The number of days to reach 70% correct answers and percent of correct responses were analyzed. All Dp(16)1/Yey, Ts1Cje and WT mice reached Stage 5 of pre-training. No differences between genotypes were found in percent of correct responses. Five Ts65Dn and one WT animals reached Stage 5 and only one Ts65Dn mouse reached VD. Ts1Cje mice took longer to move to VD vs. WT P=0.09). There were no differences between Dp(16)1/Yey and WT mice. At VD, the average percent of correct answers was significantly lower in Dp(16)1/Yey and Ts1Cje compared to WT littermates (P<0.05). These results have been submitted for publication. (3) Human fetuses with trisomy 21 (T21) have atypical brain development that is apparent sonographically in the second trimester. We hypothesized that by analyzing and integrating dysregulated gene/protein expression and pathways common to humans with DS and mouse models we could discover novel targets for prenatal therapy. We tested the safety and efficacy of apigenin, identified using this approach, in both human amniocytes from fetuses with T21 and in the Ts1Cje mouse model. In vitro, T21 cells cultured with apigenin had significantly reduced oxidative stress and improved antioxidant defense response. In vivo, apigenin treatment mixed with chow was administered prenatally to the dams and fed to the pups over their lifetimes. There was no significant increase in birth defects or pup deaths resulting from the prenatal apigenin treatment. Apigenin significantly improved several developmental milestones and spatial olfactory memory in Ts1Cje neonates. In addition, we noted novel sex-specific effects on exploratory behavior and long-term hippocampal memory in adult mice, with males showing significantly more improvement than females. We demonstrated that the therapeutic effects of apigenin are pleiotropic, resulting in decreased oxidative stress, activation of pro-proliferative and pro-neurogenic genes (KI67, Nestin, SOX2 and PAX6), reduction of the pro-inflammatory cytokines INFG, IL1A and IL12P70 through the inhibition of NFB signaling, increase of the anti-inflammatory cytokines IL10 and IL12P40 and increased expression of the angiogenic and neurotrophic factors VEGFA and IL7. These studies provide proof-of-principle that apigenin has multiple therapeutic targets in preclinical models of Down syndrome. This work has been submitted for publication.