Zugl.: Frankfurt (Main), Universiẗat, Diss., 2008.

Univ., Diss., 2009--Marburg.

Universiẗat, Diss., 2005--Frankfurt (Main).

Universiẗat, Diss., 2006--Giessen.

Zugl.: Giessen, Univ., Diss., 2006

Frankfurt (Main), Univ., Diss., 2001.

Universiẗat, Diss., 2002--Frankfurt am Main.

abstract: The need of organs for transplantation has become an increasing medical need due to a limited donor organ supply. Many organs fail within 10 years due to acute and chronic rejection. Acute or antibody mediated rejection leads to decreased long term graft survival and increases the need for a repeat transplant. In prior work, reducing endothelial heparan sulfation and blockade of chemokine-glycosaminoglycan (GAG) interaction with Myxomavirus-derived protein, M-T7, reduced aortic and renal graft vascular inflammation and rejection. Conditional endothelial Ndst1 deficiency and inhibition of chemokine-GAG interaction reduces early allograft damage and suggest new therapeutic options for graft rejection. Here acute renal rejection was examined in grafts with conditional endothelial N-deacetylase-N-sulfotransferase-1 knockout (Ndst1-/-) and in wildtype (WT) C57Bl6/J grafts treated with saline, M-T7, antisense oligonucleotides (ASO) for Ndst1 or a scrambled ASO control. Viruses have a highly adaptive ability to evade hosts defense and immune response. The immunomodulatory proteins derived from viruses provide potential therapeutic uses to alleviate this need for organs. The Myxoma virus derived protein M-T7 is a promising therapeutic for reducing kidney transplant rejection. Orthotopic transplantations in mice are extremely difficult and costly because they require a highly trained microsurgeon. This kidney to kidney subcapsular and subcutaneous transplant model is a practical and simpler method that requires fewer mice, one kidney can be used for transplants in 6 or more mice and there is much lower morbidity, pain and mortality. Heterotopic transplantation of allografts is a simple model for preliminary testing of treatments for early inflammation, ischemia, and graft rejection. Subcapsular kidney transplantation provides a first step approach to test virus-derived proteins as potential treatments to reduce transplant rejection and inflammation. This project reports on a broadly applicable platform on which to rapidly and conveniently test new treatments for transplant rejection. This finding will significantly lower the barrier to entry for labs which are interested in translating their laboratory findings to animal models of organ transplantation which is a complex surgical procedure, and thus accelerate the bench-to-bedside translation of novel, putative treatments for transplant rejection as an initial screening tool. / Dissertation/Thesis / Masters Thesis Molecular and Cellular Biology 2020

Multiple Sclerosis (MS) is a chronic, inflammatory disease of the central nervous system. MS is caused by the immune-mediated destruction of myelin and oligodendrocytes, resulting in demyelination and neurodegeneration. The microRNA miR-145-5p has been demonstrated to be upregulated in MS lesions. Our lab has previously shown that dysregulation of miR-145-5p can interfere with oligodendrocyte differentiation in mice and that knockout of miR-145-5p protects mice from experimental autoimmune encephalomyelitis (EAE), a model for MS. The objective of this study is to determine if inhibition of miR-145-5p with an antisense oligonucleotide (ASO) is sufficient to protect mice from EAE. Female mice were induced with EAE and then treated with a control or miR-145 ASO at the onset of disease. We evaluated disease progression by monitoring clinical severity, and evaluating molecular and structural characteristics of EAE by RT-qPCR, histology, immunohistochemistry and electron microscopy. We have shown that the miR-145 ASO reduced miR-145-5p expression in the lumbar spinal cord, spleen and thymus following EAE induction. Treatment with the miR-145-5p ASO resulted in improved clinical severity of EAE, reduced neuroinflammation and increased myelination. Inhibition of miR-145-5p may represent a novel treatment for MS.

Le cancer de la prostate (CaP) est actuellement le cancer le plus fréquent en France et constitue l'une des principales causes de décès par cancer chez l'homme dans les pays industrialisés. Un tiers des patients avec un CaP à priori localisé auront déjà des micro-métastases au moment du traitement local. Ces patients qui répondent dans un premier temps à la castration (hormonothérapie) seront cependant en échappement hormonal dans les 2 ans qui suivent. Récemment, plusieurs essais cliniques de phase III ont rapporté un gain de survie avec la chimiothérapie à base de docétaxel dans les CaPs métastatiques résistants à la castration. Néanmoins, la survie n'est prolongée que de 2 ou 3 mois et de nouvelles approches thérapeutiques ciblant des voies de signalisation spécifiques sont donc nécessaires. Les travaux réalisés au cours de cette Thèse ont permis tout d'abord de montrer, grâce à l'utilisation de TMAs, que la surexpression de TP53INP1, une protéine de réponse au stress, était un facteur de mauvais pronostic dans le CaP, prédictif notamment du risque de rechute biologique. Nous avons ensuite pu montrer grâce à des xénogreffes de cellules tumorales (LNCaP) que les taux d'ARNm de TP53INP1 diminuaient durant l'hormonothérapie et que TP53INP1 était de nouveau significativement surexprimée dans les tumeurs résistantes à la castration. Nous avons développé et déposé un brevet pour un oligonucléotide antisens (ASO) inhibant TP53INP1. Le traitement in vitro des lignées cellulaires hormonosensibles LNcaP et hormono-résistantes C4-2 par l'ASO induit une diminution d'expression de la protéine TP53INP1, inhibe la prolifération cellulaire et induit une augmentation de l'apoptose. / Prostate cancer (PC) is the most common malignancy in France and one of the most frequent leading causes of cancer-related death in men in industrialized countries. Even with aggressive screening, approximately one-third of patients believed to have localized PC will already have micro-metastatic disease at the time of definitive local therapy. These patients initially respond to androgen ablative therapy, but with time, their tumors ultimately become unresponsive and recur within 2 years as castration-resistant prostate cancer (CRPC). Recently, docetaxel-based regimens have shown improved survival in men with CRPC in phase III studies. However, the median overall survival was prolonged for only 2-3 months, and thus development of new therapeutic approaches that target relevant signaling pathways are essential to restore the androgen-sensitivity of CRPC. We showed, using tissue micro-array (TMA) analysis, that over-expression of Tumor Protein 53-Induced Nuclear Protein 1 (TP53INP1), a cell stress response protein, is a worse prognostic factor in PC, particularly predictive of biological cancer relapse. We also we found that TP53INP1 protein expression decreases during castration therapy (CT) and significantly increases in human CRPC. TP53INP1 mRNA was also significantly increased in castration-resistant (CR) tumors of LNCaP xenograft compared to the castration-sensitive (CS) taken before CT. We developed and world-wide patented one antisense oligonucleotide (ASO) targeting TP53INP1 (PCT/IB2011/054555). Treatment of LNCaP and C4-2 cells in vitro with TP53INP1 ASO downregulates TP53INP1 protein level, inhibits proliferation and induces apoptosis.