The placenta clearly plays a critical role in reproduction but we know very little about the factors that stimulate and control it’s early development. Work in collaboration with Professor Graham Burton (Department of PDN) is focused on understanding what these factors are, how they are controlled, how they act on the early trophoblast and how they may be perturbed in pathological pregnancies. This work involves the use of next generation sequencing and complex cell culture models to characterise the factors and investigate how they act on trophoblast derived cells. This work is funded by a programme grant from the Medical Research Council. This work is a continuation of a longstanding collaboration in which we have identified the importance of oxidative and endoplasmic reticulum stress in the placenta; (see papers by Yung et al, 2014 and Cindrova-Davies et al, 2011 for examples). We are also interested in the relationship between capillary and villus growth in the placenta and use 3D image reconstruction to investigate this.
Mouse Models of Clear Cell Ovarian Cancer
Ovarian cancer is the fifth most common cancer in women in the United Kingdom and one sub-type is the so-called clear cell carcinoma. This tumour type is an uncommon (approximately 10% of all ovarian carcinomas) and is actually thought to originate in the endometrial cells within the uterus. The type of cancer has a poor prognosis mostly due to late diagnosis.
There is evidence of an association between endometriosis and clear cell ovarian cancer; (endometriosis is a benign gynaecological condition in which endometrial tissue grows outside the uterus, commonly within the peritoneal cavity). There is also some evidence that mutations in endometriotic cells may lead to the development of clear cell ovarian cancer. Some of the work in my laboratory is focused on specifically exploring this question as we have developed a mouse model of endometriosis, which allows genetic manipulation of the endometrial cells and also of the host environment. We are able to introduce specific mutations or knockdown specific genes in the endometrial tissue and then determine whether this alters the growth of the endometriotic tissue and whether this tissue progresses towards clear cell ovarian cancer. Techniques used in these projects involve the generation and use of genetically modified mice, lentiviral transduction to either knockdown or over express modified genes, in vivo imaging of mice as they develop lesions (see photograph to the left for an example of this). Some of the analytical tools we use to investigate lesion development include next generation sequencing (particularly RNA-Seq and ChIP-Seq). This work is funded by CRUK and STEPS. This work is carried out in close collaboration with Dr James Brenton.
The role of Paroxonase 3(PON3) in Development
We previously identified the paroxonase 3 gene as one being acutely up regulated in both the lung and intestine in the late stages of gestation (Belteki et al, 2010). We also showed that knocking out this gene led to embryonic lethality (Kempster et al, 2011). We have a programme of work to define why deletion of this gene causes embryonic loss. We have investigated the presence of the protein in early mouse embryos cultured in vitro. Immuno localisation indicates that the protein begins to appear in the late blastocyst and only in the trophectoderm cells. (For example see the picture below, which illustrates such an embryo). In embryos that have implanted there is further expression also in trophectoderm-derived cells. Other aspects of this project involves the generation of a conditional knockout which will allow us to specifically ablate the Pon3 at selected times during development and also in specific tissues postnatally. This work is supported by a MRC project grant.
A key feature of my research is the combination of genetically manipulable models of reproductive processes and the study of relevant clinical human material. Particularly exciting is the use of next generation sequencing methods in the POPS cohort. I therefore co-supervise with Professor Smith several projects seeking to exploit these technologies in the investigation of fundamental questions of clinical relevance using samples collected from this study. Some of the analytical tools we use to investigate placental development and pathology include next generation sequencing – particularly RNA-Seq (mRNA, ncRNA and miRNA) and WGBS.
Recent Key Publications
King CM, Barbara C, Prentice A, Brenton JD and Charnock-Jones DS (2015). Models of endometriosis and their utility in studying progression to ovarian clear cell carcinoma. J. Pathol. doi: 10.1002/path.4657 [Epub ahead of print]
Yung HW, Atkinson D, Campion-Smith T, Olovsson M, Charnock-Jones DS, and Burton GJ (2014). Differrential activation of placental unfolded protein response pathways implies heterogeneity in causation of early- and late-onset pre-eclampsia. J. Pathology. 234(2): 262-276. DOI:10.1002/path.4394.
Yung HW, Colleoni F, Atkinson D, Cook E, Murray A, Burton G, and Charnock-Jones DS (2014). Influence of speed of sample processing on placental energetics and signalling pathways: implications for tissue collection. Placenta. 35(2): 103-108.
Staff AC, Benton SJ, von Dadelszen P, Roberts JM, Taylor RN, Powers RW, Charnock-Jones DS, and Redman CWG (2013). Redefining preeclampsia using placenta-derived biomarkers. Hypertension. 61(5): 932-942
Yung HW, Hemberger M, Watson ED, Senner CE, Jones CP, Kaufman RJ, Charnock-Jones DS, and Burton GJ (2012). Endoplasmic reticulum stress disrupts placental morphogenesis: implications for human intrauterine growth restriction. J. Path. 228: 554-564. DOI:10.1002/path.4068
Tattersall M, Cordeaux Y, Charnock-Jones DS, and Smith GCS (2012). Prolonged stretch of human myometrium increases gastrin releasing peptide (GRP) expression and GRP receptor antagonists inhibit contractility. J. Physiol. 590(Pt 9): 2081-2093. DOI: 10.1113/jphysiol.2012.228239
Cindrova-Davies T, Sanders DA, Burton GJ, and Charnock-Jones DS (2011). Soluble FLT1 sensitizes endotehlial cells to inflammatory cytokines by antagonizing VEGF receptor-mediated signalling. Cardiovascular Research. 89: 671-679
Hurley D, Araki H, Tamada Y, Dunmore BJ, Sanders DA, Humphreys S, Affara M, Imoto S, Yasuda K, Tomiyasu Y, Tashiro K, Savoie C, Cho V, Smith S, Kuhara S, Miyano S, Charnock-Jones DS, Crampin EJ, and Print CG (2011). Gene network inference and visualization tools for biologists: application to new human transcriptome datasets. Nucleic Acids Research. DOI:10.1093/nar/gkr902
Kempster SL, Belteki G, Licence D, Charnock-Jones DS, and Smith GC (2011). Disruption of paraoxinase 3 impairs proliferation and antioxidant defenses in human A549 cells and causes embryonic lethality in mice. Am. J. Physiol. Endocrin Metab. DOI:10.1152/ajpendo.00357.2011
Cheng C-W, Licence D, Cook E, Luo F, Arends MJ, Smith SK, Print CG, and Charnock-Jones DS (2011). Activation of mutated K-ras in donor endometrial epithelium and stroma promotes lesion growth in an intact immunocompetent murine model of endometriosis. J. Pathol. 224(2): 261-269
Kempster SL, Belteki G, Forhead AJ, Fowden AL, Catalano RD, Lam BY, McFarlane I, Charnock-Jones DS, and Smith GC (2011). Developmental control of the Nlrp6 inflammasome and a substrate, IL-18, in mammalian intestine. Am. J. Physiol Gastrointest Liver Physiol. 300: G253-G263
Yung HW, Charnock-Jones DS, and Burton GJ (2011). Regulation of AKT Phosphorylation at Ser473 and Thr308 by Endoplasmic Reticulum stress modulates substrate specificity in a severity dependent manner. PLoS One. 6: e17894
Affara M, Dunmore BJ, Sanders DA, Johnson N, Print CG, and Charnock-Jones DS (2011). MMP1 bimodal expression and differential response response to inflammatory mediatore is linked to promoter polymorphisms. BMC Genomics. 12: 43
Yamaji M, Bielby H, Licence D, Cheng CW, Cook E, Smith SK, Print CG, and Charnock-Jones DS (2010). VEGF-A loss in the haematopoietic and endothelial lineages exacerbates age-induced renal changes. Microvasc Res. 80: 372-383
Mokhtar NM, Cheng CW, Cook E, Bielby H, Smith SK, and Charnock-Jones DS (2010). Progestin regulates chemokine (C-X-C motif) ligand 14 transcript level in human endometrium. Mol. Hum. Reprod. 16: 170-177
Belteki G, Kempster SL, Forhead AJ, Giussani DA, Fowden AL, Curley A, Charnock-Jones DS, and Smith GC (2010). Paraoxonase-3, a putative circulating antioxidant, is systemically up-regulated in late gestation in the fetal rat, sheep, and human. J. Clin Endocrinol. Metab. 95: 3798-3805
Yung HW, Korolchuk S, Tolkovsky AM, Charnock-Jones DS, and Burton GJ (2007). Endoplasmic reticulum stress exacerbates ischemia-reperfusion-induced apoptosis through attenuation of Akt protein synthesis in human choriocarcinoma cells. FASEB J. 21: 872-884
Cindrova-Davies T, Yung HW, Johns J, Spasic-Boskovic O, Korolchuk S, Jauniaux E, Burton GJ, and Charnock-Jones DS (2007). Oxidative stress, gene expression and protein changes induced in th ehuman placenta during labor. Am J. Pathol. 171: 1168-1178
Cindrova-Davies T, Spasic-Boskovic O, Jauniaux E, Charnock-Jones DS, and Burton GJ (2007). Nuclear factor-kappa B, p38, and stress-activated protein kinase mitogen-activated protein kinase signaling pathways regulate proinflammatory cytokines and apoptosis in human placental explants in response to oxidative stress: effects of antioxidant vitamins. Am J Pathol. 170(5): 1511-1520
Cheng CW, Bielby H, Licence D, Smith SK, Print CG, and Charnock-Jones DS (2007). Quantitative cellular and molecular analysis of the effect of progesterone withdrawal in a murine model of decidualization. Biol Reprod. 76(5): 871-883