抗原变异可使得多种致病微生物易于逃避宿主免疫应答。表达在感染红细胞表面的恶性疟原虫红细胞表面蛋白1（PfPMP1）与感染红细胞、内皮细胞、树突状细胞以及胎盘的单个或多个受体作用，在黏附及免疫逃避中起关键的作用。每个单倍体基因组var基因家族编码约60种成员，通过启动转录不同的var基因变异体为抗原变异提供了分子基础。

疟原虫var基因转换速率变化导致抗原变异[英]／Paul H, Robert P, Christodoulou Z, et al//Proc Natl Acad Sci U S A

Background: Currently, a lack of consensus exists on how best to perform and interpret quantitative real-time PCR (qPCR) experiments. The problem is exacerbated by a lack of sufficient experimental detail in many publications, which impedes a reader’s ability to evaluate critically the quality of the results presented or to repeat the experiments.

Content: The Minimum Information for Publication of Quantitative Real-Time PCR Experiments (MIQE) guidelines target the reliability of results to help ensure the integrity of the scientific literature, promote consistency between laboratories, and increase experimental transparency. MIQE is a set of guidelines that describe the minimum information necessary for evaluating qPCR experiments. Included is a checklist to accompany the initial submission of a manuscript to the publisher. By providing all relevant experimental conditions and assay characteristics, reviewers can assess the validity of the protocols used. Full disclosure of all reagents, sequences, and analysis methods is necessary to enable other investigators to reproduce results. MIQE details should be published either in abbreviated form or as an online supplement.

Summary: Following these guidelines will encourage better experimental practice, allowing more reliable and unequivocal interpretation of qPCR results.

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The MIQE Guidelines: Minimum Information for Publication of Quantitative Real-Time PCR Experiments

Stem cell biology has come of age. Unequivocal proof that stem cells exist in the haematopoietic system has given way to the prospective isolation of several tissue-specific stem and progenitor cells, the initial delineation of their properties and expressed genetic programmes, and the beginnings of their utility in regenerative medicine. Perhaps the most important and useful property of stem cells is that of self-renewal. Through this property, striking parallels can be found between stem cells and cancer cells: tumours may often originate from the transformation of normal stem cells, similar signalling pathways may regulate self-renewal in stem cells and cancer cells, and cancer cells may include 'cancer stem cells' - rare cells with indefinite potential for self-renewal that drive tumorigenesis.

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Stem cells, cancer, and cancer stem cells

Mutations of the gene Lps selectively impede lipopolysaccharide (LPS) signal transduction in C3H/HeJ and C57BL/10ScCr mice, rendering them resistant to endotoxin yet highly susceptible to Gram-negative infection. The codominant Lpsd allele of C3H/HeJ mice was shown to correspond to a missense mutation in the third exon of the Toll-like receptor-4 gene (Tlr4), predicted to replace proline with histidine at position 712 of the polypeptide chain. C57BL/10ScCr mice are homozygous for a null mutation of Tlr4. Thus, the mammalian Tlr4 protein has been adapted primarily to subserve the recognition of LPS and presumably transduces the LPS signal across the plasma membrane. Destructive mutations of Tlr4 predispose to the development of Gram-negative sepsis, leaving most aspects of immune function intact.

http://science.sciencemag.org/content/sci/282/5396/2085.full.pdf

Defective LPS Signaling in C3H/HeJ and C57BL/10ScCr Mice: Mutations in Tlr4 Gene

Defective LPS signaling in C3 H/HeJ and C57 BL/10 ScCr mice: Mutations in Tlr4 Gene

Targeted disruption of Sonic hedgehog reveals an essential role in hair follicle morphogenesis

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Erratum: Cellular origin of bladder neoplasia and tissue dynamics of its progression to invasive carcinoma (Nature Cell Biology (2014) 16 (469-478))

Developmental biology: Drosophila proteins pave the way to gene mutations

The present invention provides transcription factors associated with the hedgehog signaling pathway that are regulated by dephosphorylation by phosphatases. Hedgehog response elements (HRE) that interact with the dephosphorylated transcription factors are also provided as well as methods for identifying compounds that are phosphatase inhibitors. Methods of treating tumors in a subject by modulating the phosphorylation of the transcription factor are also included.

Method for identifying an agent that affects a hedgehog signaling pathway

We reported that transcriptional regulation by the major Hedgehog (HH) pathway effector GLI2 in mammary gland stromal cells coordinates a hormone-responsive niche signaling program that directs epithelial stem cell activity during puberty [Zhao, et al., Science. 2017 Apr 21;356(6335)]. HH signaling is a key growth pathway in human carcinogenesis. The first targeted therapies aimed at G-protein coupled receptor Smoothened (SMO) resulted in rapid acquired resistance, underscoring the need for developing new HH pathway-targeting therapies downstream of SMO. In BC GLI-dependent transcription may involve a non-receptor-based mechanism of GLI2 activation involving cross-talk from other signaling pathways such as TGFs54, PI3K55,56, Wnt, or NF-κB. Accordingly, we investigated the role of HH pathway and its transcriptional effectors using the MMTV-PyMT BC mouse model. Development of mammary tumors arising proceeds through neoplastic lesions ranging from carcinoma in situ (10-12 weeks) to highly invasive ductal carcinoma with high incidence of pulmonary metastasis at 16-18 weeks. Epithelial ablation of GLI2 dramatically reduced tumor progression. Cre recombinase under control of the Ck14 promoter to genetically ablate GLI2 in basal cells of the mammary gland thus dramatically attenuated metastasis despite continued (albeit reduced) formation of primary tumors. In human BC, we applied Bayesian methods to gene expression data to identify metastatic BC with HH pathway activation. Kaplan-Meier analysis demonstrated significantly worse progression-free survival in patients with HH pathway activity (Log-rank p = 0.0013). Next, we analyzed 1294 BC samples, stratified according to HH activity, using the HH probability as a continuous score ranging between 0 and 1, univariate Cox regression analysis supports the hypothesis that HH activity in BC is a risk factor for relapse after surgery, HR = 2.45 (95% CI: 1.67 – 3.61, p = 2.72e-6). There is also a significant difference in survival between HH-actives and HH-inactives (Log-rank p = 3.91e-4), suggesting a pathogenic role of GLI activation in BC progression and metastasis. Arsenic trioxide (ATO), inhibits HH pathway activity by de-stabilizing the GLI transcriptional effectors of HH signaling, likely due to arsenic displacement of zinc within the DNA-binding zinc fingers of GLI proteins. ATO treatment of BC cell lines resulted in dose-dependent cell growth inhibition (Alamar Blue) and induction of apoptosis (PARP cleavage and Annexin V expression) at clinically achievable concentrations. In NSG mice with orthotopic transplant of 3e5 SU151 human BC PDX, with strongly positive GLI-active signature, daily IP injection of 10mg/kg ATO resulted in marked tumor growth inhibition in vivo (2.75-fold smaller tumor diameter vs. PBS vehicle controls, P= 0.006; 55d vs. 17d to reach 1.5 cm criterion for ATO vs. vehicle). We conclude the major HH pathway transcriptional effector GLI2 coordinates a transcriptional program with a central role in BC growth and metastasis in a significant subset of BC, and that systemic treatment with ATO (new oral formulation in commercial development), will reduce metastatic progression and improve clinical outcomes in patients whose tumors harbor a GLI-active transcript signature. Citation Format: Beachy P, Cai S, Ma Y, Hatakeyama J, Zhao C, Stoffels M, Verhaegh W, van de Stolpe A, Pegram M. Targeted disruption of transcriptional effector GLI2 attenuates breast tumor growth and metastasis [abstract]. In: Proceedings of the 2017 San Antonio Breast Cancer Symposium; 2017 Dec 5-9; San Antonio, TX. Philadelphia (PA): AACR; Cancer Res 2018;78(4 Suppl):Abstract nr P5-03-11.

Philip A. Beachy

Papers

Targeted disruption of Sonic hedgehog reveals an essential role in hair follicle morphogenesis

Erratum: Cellular origin of bladder neoplasia and tissue dynamics of its progression to invasive carcinoma (Nature Cell Biology (2014) 16 (469-478))

Developmental biology: Drosophila proteins pave the way to gene mutations

Method for identifying an agent that affects a hedgehog signaling pathway

Abstract P5-03-11: Targeted disruption of transcriptional effector GLI2 attenuates breast tumor growth and metastasis