Targeting Echinococcus multilocularis stem cells by inhibition of the Polo-like kinase EmPlk1.
Targeting Echinococcus multilocularis stem cells by inhibition of the Polo-like kinase EmPlk1.
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DOI：
10.1371/journal.pntd.0002870
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发表时间：
2014-06
影响因子：
3.8
通讯作者：
Brehm K
中科院分区：
文献类型：
--
作者：
Schubert A;Koziol U;Cailliau K;Vanderstraete M;Dissous C;Brehm K
泡状包虫病 (AE) 是一种由多房棘球绦虫幼虫引起的危及生命的疾病。 AE 病理学的关键是寄生虫后绦虫阶段的持续浸润性生长，这是由称为生发细胞的体细胞干细胞群驱动的。目前使用苯并咪唑的抗 AE 化疗无法有效消除萌发细胞群，从而导致治疗停止后寄生虫生长得到缓解。我们在此描述了由基因 emplk1 编码的 EmPlk1 的特征，该基因与调节真核细胞有丝分裂的 Polo 样激酶的 Plk1 亚家族成员具有显着的同源性。我们通过 RT-PCR、转录组学和原位杂交证明了 emplk1 的萌发细胞特异性表达。我们还表明，当在非洲爪蟾卵母细胞中异源表达时，EmPlk1 可以诱导生发囊泡破裂，表明它是一种活性激酶。 BI 2536 是一种 Plk1 抑制剂，已在抗癌临床试验中进行了测试，这种活性在 BI 2536 存在下显着受到抑制。添加浓度低至 20 nM 的 BI 2536 可显着阻止培养的棘球蚴萌发细胞中后绦虫囊泡的形成。此外，低浓度的 BI 2536 在体外消除了成熟后绦虫囊泡中的萌发细胞群，产生了不再能够增殖的寄生虫组织。我们得出的结论是，BI 2536 通过直接抑制 EmPlk1，在体外有效灭活寄生虫幼虫中的多房艾美耳球虫萌发细胞，从而诱导有丝分裂停滞和萌发细胞死亡。由于生发细胞对于宿主内寄生虫的增殖和转移形成具有决定性作用，因此 BI 2536 和相关化合物是非常有前景的化合物，可在 AE 化疗中补充苯并咪唑。致命性疾病AE的特征是多房绦虫的后绦虫幼虫在宿主器官内持续浸润性生长。这种类似癌症的进展完全是由一群寄生虫干细胞（萌发细胞）驱动的，必须消除这些干细胞才能有效治愈该疾病。目前使用苯并咪唑的治疗方案仅能抑制寄生虫，因此显然不能有效杀死生殖细胞。我们在此描述了一种新型的、可药物化的寄生虫酶 EmPlk1，它特异性调节萌发细胞增殖。我们发现，最初设计用于抑制 EmPlk1 人类直系同源物的化合物 BI 2536，也可以在低剂量下抑制寄生虫蛋白。此外，低剂量的 BI 2536 在体外消除了棘球绦虫幼虫的萌发细胞，并阻止了寄生虫的生长和发育。我们认为 BI 2536 和相关化合物是有前途的药物，可以补充当前的苯并咪唑治疗，以实现寄生虫杀灭。
Alveolar echinococcosis (AE) is a life-threatening disease caused by larvae of the fox-tapeworm Echinococcus multilocularis. Crucial to AE pathology is continuous infiltrative growth of the parasite's metacestode stage, which is driven by a population of somatic stem cells, called germinative cells. Current anti-AE chemotherapy using benzimidazoles is ineffective in eliminating the germinative cell population, thus leading to remission of parasite growth upon therapy discontinuation. We herein describe the characterization of EmPlk1, encoded by the gene emplk1, which displays significant homologies to members of the Plk1 sub-family of Polo-like kinases that regulate mitosis in eukaryotic cells. We demonstrate germinative cell-specific expression of emplk1 by RT-PCR, transcriptomics, and in situ hybridization. We also show that EmPlk1 can induce germinal vesicle breakdown when heterologously expressed in Xenopus oocytes, indicating that it is an active kinase. This activity was significantly suppressed in presence of BI 2536, a Plk1 inhibitor that has been tested in clinical trials against cancer. Addition of BI 2536 at concentrations as low as 20 nM significantly blocked the formation of metacestode vesicles from cultivated Echinococcus germinative cells. Furthermore, low concentrations of BI 2536 eliminated the germinative cell population from mature metacestode vesicles in vitro, yielding parasite tissue that was no longer capable of proliferation. We conclude that BI 2536 effectively inactivates E. multilocularis germinative cells in parasite larvae in vitro by direct inhibition of EmPlk1, thus inducing mitotic arrest and germinative cell killing. Since germinative cells are decisive for parasite proliferation and metastasis formation within the host, BI 2536 and related compounds are very promising compounds to complement benzimidazoles in AE chemotherapy. The lethal disease AE is characterized by continuous and infiltrative growth of the metacestode larva of the tapeworm E. multilocularis within host organs. This cancer-like progression is exclusively driven by a population of parasite stem cells (germinative cells) that have to be eliminated for an effective cure of the disease. Current treatment options, using benzimidazoles, are parasitostatic only, and thus obviously not effective in germinative cell killing. We herein describe a novel, druggable parasite enzyme, EmPlk1, that specifically regulates germinative cell proliferation. We show that a compound, BI 2536, originally designed to inhibit the human ortholog of EmPlk1, can also inhibit the parasite protein at low doses. Furthermore, low doses of BI 2536 eliminated germinative cells from Echinococcus larvae in vitro and prevented parasite growth and development. We propose that BI 2536 and related compounds are promising drugs to complement current benzimidazole treatment for achieving parasite killing.
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DOI：
10.1590/s0001-37652011000200022
发表时间：
2011-06-01
期刊：
Anais da Academia Brasileira de Ciências
影响因子：
--
作者：
Dissous, Colette;Grevelding, Christoph G;Long, Thavy
通讯作者：
Long, Thavy
影响因子：
5.8
作者：
Gelmedin, Verena;Caballero-Gamiz, Rocio;Brehm, Klaus
通讯作者：
Brehm, Klaus
影响因子：
4
作者：
Gelmedin, Verena;Spiliotis, Markus;Brehm, Klaus
通讯作者：
Brehm, Klaus
影响因子：
3.7
作者：
Long T;Vanderstraete M;Cailliau K;Morel M;Lescuyer A;Gouignard N;Grevelding CG;Browaeys E;Dissous C
通讯作者：
Dissous C
影响因子：
5.2
作者：
Labbe, Roselyne M.;Irimia, Manuel;Currie, Ko W.;Lin, Alexander;Zhu, Shu Jun;Brown, David D. R.;Ross, Eric J.;Voisin, Veronique;Bader, Gary D.;Blencowe, Benjamin J.;Pearson, Bret J.
通讯作者：
Pearson, Bret J.