Prostate cancer: The impact of inverse expression of the anti-apoptotic Trap 1 and pro-apoptotic DNaseI genes on prognosis and therapy resistance
Regulation and impact of DNaseI and Trap 1 in lupus nephritis and prostate cancer – Approaching causal therapy modalities
Silencing of renal nuclease DNaseI gene in lupus nephritis promotes severe progression of the disease since renal chromatin will no longer be degraded. Instead chromatin is accumulated in glomerular membranes where they bind anti-chromatin antibodies. We want to determine if silencing of DNaseI gene up-regulate Trap 1 in autoimmunity and cancer.
We have studied and successfully described the impact of an acquired loss of renal DNaseI mRNA and enzyme activity as a profound pathological event in progression of lupus nephritis into an irreversible state with complete loss of renal function. This is original research performed by the Rekvig Group at UiT/UNN. Silencing of the renal nuclease DNaseI gene in lupus nephritis promotes severe progression of the disease since renal chromatin will no longer be degraded. Instead chromatin is accumulated in glomerular membranes where they bind anti-chromatin antibodies. This promotes severe renal inflammation. In order to understand pathogenic and prognostic effect and regulation of DNaseI we will perform 5 sets of analyses: 1. To determine if a constitutively expressed transgene, the DNaseI, will protect against progressive lupus nephritis. 2. To determine if the lupus nephritis process in transplanted kidneys on a totally different genetic background is identical to lupus nephritis in autologous kidneys. 3. To determine if an abrupt silencing of the renal DNaseI gene correlates in time with up-regulation of Trap 1 because the assumed inverted expression profiles are in agreement with transcriptional interference of the two genes that overlap in their 3’- untranslated regions (UTR). We will in this project use our insight into studies of transcriptional interference to analyse regulation of Trap 1 and DNaseI in autoimmunity and cancer. 4. To determine if DNaseI and Trap1 expression levels are inversely correlated with each other in prostate cancer. This is important to establish since high Trap, and corresponding low DNaseI give the cancer a highly unfavourable prognosis. 5. To analyse regulation pathways for DNaseI and Trap 1. The Dnasel gene is localized on chromosome 16. Another gene, the TNF receptor-associated protein 1 (Trap 1), is encoded the opposite direction of Dnasel on the opposite strand, and the two transcripts overlap in their 3'- untranslated regions (UTRs) (see (20) for gene maps and (21) information on Dnasel in general). Since Trap 1 is up-regulated during stress, it has the function of a survival protein, and belongs to the heat shock protein (HSP) family (21-23). This means that Dnase 1 and Trap l genes cannot he transcribed at the same time, and the annotated transcripts mutually exclude each other. Data so far demonstrate in tissue micro-array of 12000 prostate biopsies an inverse relationship bweteen Expression of DNase I and Trap 1. In the biopsies with the highest Trap 1 expression, the biopsies express more aggressive cancer. Similarly, in prostate cancer cells, stimulation of cells with cytokines may up-regulate DNaseI and inhibit Trap 1 Expression (studies in progress). This is a phenomenon that may be used therapeutically. Due to maternal leave, the post doc has worked only 11 months yet. Therefore, we still have not the data needed to publish, but the project is in good progression. There is no relevant publication for 2016. I will in the end, thank HN for support and challenges. I am thankful for what you have done for me and for my science. I retire March 1, 2017 Ole Petter Rekvig
The impact of DNaseI and Trap 1 on prognosis and therapy resistance in prostate cancer
We have demonstrated that up-regulation of the anti-apoptotic survival protein Trap 1 is promoting progression of lupus nephritis by suppressing the pro-apoptotic endonuclease DNaseI. Trap 1 has also been shown to upregulate survival genes in cancer cells. In this project we will determine regulation pathways of Trap 1/DNaseI.
Dhivya Thiygarajan, PhD, started as post doc May 1 2014, and left for maternal leave February 13, 2015. During this time, she finished her baseline studies on basic regulatory aspects of DNaseI gene expression, and developed the methods and analytical strategies for studies of DNaseI and Trap 1 expression in prostate cancer cells. These methods were validated in tubular cells, the cells best investigated in our laboratory. She did the following observations: Proinflammatory TNFalpha upregulates DNaseI expression and another proinflammatory cytokine, the IL-1beta. We demonstrated that IL-1beta is responsible for nuclear translocation of DNaseI in an endonuclease inactive form. These data are now published. Detailed analyses demonstrated that this form of DNaseI bound to the promoter region of the Fas Receptor, which induced upregulation of Fas Receptor protein. This process make the cells susceptible for apoptosis (manuscript ready for submission). In this situation, Trap 1 is down-regulated. In parallel, we investigated the expression profiles of DNaseI and Trap 1 in prostate cancer biopsies in collaboration with Sarah Minner in Hamburg (Eppendorf Hospital) and Ian Mills at the Norwegian Center for Molecular Mesicin (NCMM, University of Oslo). The data are very promising as we consistently observed that DNaseI and Trap 1 demonstrated inverse expression profiles, and that high Trap 1 conferred to low DNaseI, and vice versa. Furthermore, high Trap 1 correlated with poor prognosis of cancer prostate, while high DNaseI correlated with a more favorable prognosis. In prostate cancer cells, we are transferring the protocols used for baseline studies in tubular cells, to determine how we can upregulate DNaseI and consequently downregulate Trap 1 and thereby improve the prognosis and therapy sensitivity of prostate cancer. This is the status of the project after 10 months studies when Dhivya Thyigarajan left for maternal leave. So far, the project is quite succesful, with new, potentially important result, particularly concerning up-regulated DNaseI as a transcription factor for the apoptosis-associated Fas Receptor. The project follows, and will follow, the plans described in the project application.
Prostate cancer: Impact of inverse expression of the anti-apoptotic Trap 1 and pro-apoptotic DNaseI genes on prognosis
Nyere data har vist at Trap 1 ekspresjon i prostatakreft celler medfører dårlig prognose og terapiresistens. Vi har utviklet innsikt i regulering av Trap 1 gjennom våre studier i lupus nefritt, hvor også Trap 1 spiller en rolle. Vi utfører en komparativ studie av genekspresjon i prostatakreft ved analyser av biopsier og prostatakreft cellelinjer
Prosjektet startet April 2014. I henhold til planen beskrevet i søknaden har vi i 2014 oppnådd følgende: Vi har mottatt prostatakreft cellelinjer fra Ian Mills, UiO, og Norsk Center for Molekylærmedisin. Disse er klare til å implementeres i forsøk. Vider har vi i samarbeid med Sarah Minner ved Eppendorf sykehus, Hamburg, allerede analysert prostatabiopsier for ekspresjon av DNaseI og Trap 1. Vi analyserte rundt 12000 biopsier i form av Tissue microarray (TMA). Denne delstudien var svært omfattende og viste at det generelt var et inverst ekpresjonsmønster for DNaseI og Trap 1 som anført som hypotese for prosjektet. Videre viste studien at høy Trap 1 ekspresjon var relatert til dårlig prognose. Vi er nå klare til å analysere ekspresjon av disse to genene i cellelinjer som ledd i studier av hvordan disse to genene reguleres. Vi utfører disse forsøkene etter prosedyrer vi har utviklet for å studere reguleringen av DNaseI og Trap 1 i lupus nefritt, samt å videreutvikle kunnskapen om DNaseI og Trap 1 sine biologiske funksjoner. Særlig interessant er studien av DNaseI translokasjon til cellekjernene ved inflammasjon. Her har vi oppdaget en helt ny funksjon til enzymet. Vi har og deltatt i full mRNA profilering i prostatacellelinjer. Denne databasen er klar til analyser av presumptive signalveier for regulering av DNaseI og Trap 1 Prosjektet har fått en god oppstart, men vil bli noe forsinket da post doc Dhivya Thiyagarajan nettop har gått ut i svangerskapspermisjon (fra 28. januar 2015). Allikevel regner vi med at prosjektet vil ha en viss progresjon også i 2015. På grunn av den korte funksjonstiden fra oppstart er det ikke skrevet noen artikkel enda.
TNFa Amplifies DNaseI Expression in Renal Tubular Cells while IL-1ß Promotes Nuclear DNaseI Translocation in an Endonuclease-Inactive Form.
PLoS One 2015;10(6):e0129485. Epub 2015 jun 11
Murine and Human Lupus Nephritis: Pathogenic Mechanisms and Theoretical Strategies for Therapy.
Semin Nephrol 2015 Sep;35(5):427-38.