| 研究課題/領域番号 |
21K09324
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| 研究種目 |
基盤研究(C)
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| 配分区分 | 基金 |
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| 応募区分 | 一般 |
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| 審査区分 |
小区分56020:整形外科学関連
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| 研究機関 | 大分大学 |
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研究代表者 |
カーン シャキル 大分大学, 医学部, 助教 (70746867)
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| 研究期間 (年度) |
2021-04-01 – 2024-03-31
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| 研究課題ステータス |
交付 (2022年度)
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| 配分額 *注記 |
4,160千円 (直接経費: 3,200千円、間接経費: 960千円)
2023年度: 650千円 (直接経費: 500千円、間接経費: 150千円)
2022年度: 1,690千円 (直接経費: 1,300千円、間接経費: 390千円)
2021年度: 1,820千円 (直接経費: 1,400千円、間接経費: 420千円)
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| キーワード | Spina Bifida Aperta / Chicken model / Mammalian model / Prosaposin / Neurotrophic factor / Therapeutic potentiality / 二分脊椎 / 哺乳類モデル / プロサポシン / 神経成長因子 / 治療 / ラット / マウス |
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| 研究開始時の研究の概要 |
１）レチノイン酸(60mg/kg)を妊娠ラットに飲ませ、二分脊椎を発症させる。
２）1群にPS18(0.2mg, 2mg/kg)を投与、別群に無意味な配列のペプチドを投与する。
３）妊娠21日に麻酔後に胎児をホルマリン固定し、両群の発症率を比較する。
４）二分脊椎の患部の実体顕微鏡写真を撮り、パラフィン包埋する。
５）パラフィン切片を作製し、治療効果の免疫組織学的な検討を行う。
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| 研究実績の概要 |
Spina bifida aperta (SBA), a developmental disorder of the spinal cord, causes high pediatric mortality and is the leading cause of lifelong neurological complications including paralysis, orthopedic abnormalities, problems with bladder and bowel control, and painful complications. Current therapeutics for SBA have various limitations, including incomplete preservation of spinal tissue and the inability to rescue sensorimotor functions. We found that intra-amniotic delivery of prosaposin-derived 18 mer-peptide (PS18), a potential neurotrophic factor, to the chick embryo with SBA enhances the regeneration process and protects spinal tissue in developing spinal cord. Furthermore, treatment with PS18 dramatically improved the walking and bowel movement ability and reduce and painful complications of hatched chicks in postnatal life. The results are published as first author in a highly reputed scientific journal (Khan et al., iScience 2023; IF: 6.107).
In order to aim for clinical application of PS18, verification using not only the chicken model but also the mammalian SBA model is necessary. In this regard, we are trying to develop a mice model of SBA by inducing Zika virus which causes neural tube malformations and other birth defects. Zika virus solution (1x10^4 PFU in 200μL PBS) was administered subcutaneously to STAT1 KO mice on the 6th day of pregnancy to induce SBA and/or other defects in the fetus. Once a mice model with SBA or other neurological birth defects is developed, we will evaluate the therapeutic efficacy of PS18 with the aim of writing papers.
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| 現在までの達成度 (区分) |
現在までの達成度 (区分)
2: おおむね順調に進展している
理由
Due to the effects of COVID-19 pandemic, the use of animal testing facilities and laboratories was restricted in earlier years. However, currently, the experiments are being conducted as planned.
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| 今後の研究の推進方策 |
In FY2023, we will proceed with the development of a mammalian SBA model by Zika virus infection or administration of retinoic acid. In the Zika virus infection-induced SBA model, Asian type of Zika virus is used in transgenic mice (STAT1 KO mice). Pregnant mice are given 1x10^4 PFU of Zika virus in 200μL PBS at embryonic day 6. In the SBA model with retinoic acid, retinoic acid (dissolved in olive oil) is administered orally at 60 mg/kg to pregnant animals when the embryo is 10 days old in the uterus of mice/rat.
Morphological, histological, and behavioral methods will be used to confirm the establishment of the SBA model. Congenital Zika virus infection has also been associated with other birth defects, including but not limited to brain atrophy and asymmetry, abnormally formed or absent brain structures, hydrocephalus, and neuronal migration disorders. Therefore, we will assess Zika virus infection-induced any of these birth defects in fetus. Next, we will assess the therapeutic efficacy of PS18 to treat SBA and other birth defects using these mammalian models.
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