This thesis have explored the functions of the protein Tissue Factor (TF), which together with its ligand coagulation factor VII/VIIa (FVII/FVIIa) forms a proteolytic complex that functions in initiation of blood coagulation and activation of cell signaling. In paper I, the mechanisms behind the observation that TF/FVIIa signaling protects cells from apoptosis were further investigated. Using cell culture models, we found that antiapoptotic signaling by TF/FVIIa requires signaling by the Insulin-like growth factor I receptor (IGF-1R), as synthetic IGF-1R inhibitors and IGF1-R siRNA knock-down abolished the antiapoptotic effect of FVIIa. Furthermore, the IGF-1R translocated to the cell nucleus after FVIIa stimulation, implying a role in regulation of gene expression. Papers II and III describe the discovery that the Eph tyrosine kinase receptors EphB2 and EphA2 are proteolytically cleaved directly by TF/FVIIa. By using mass spectrometry and N-terminal Edman sequencing, the exact cleavage site was identified after a conserved arginine residue in the EphA2/EphB2 ligand binding domains, in agreement with the cleavage preferences of FVIIa. TF and EphA2/EphB2 co-localized in cancer cell lines and FVIIa potentiated ligand-dependent Eph signaling by increasing cytoskeletal remodeling and cell repulsion, demonstrating a novel proteolytical event that modulates Eph receptor signaling. In paper IV, expression of TF was investigated in colorectal cancer in both the stromal and tumor cell compartments by immunohistochemistry using an anti-TF-antibody developed and validated by the Human Protein Atlas project. In normal large intestine, TF was strongly expressed in the innermost pericryptal sheath cell layer lining the epithelium, in a cell population distinct from intestinal pericryptal myofibroblasts. We evaluated TF expression in two colorectal cancer materials, and found that TF was variably present in both the stromal and tumor cell compartments. TF expressed by pericryptal sheath cells was progressively lost after the adenoma-to-carcinoma transition and was a strong predictor of survival in rectal but not colon cancer patients independently of disease stage, histological tumor grade and age. In summary, this thesis demonstrates novel signaling mechanisms for the TF/FVIIa complex, and provides evidence of a hitherto unknown role of TF expressed by a specific population of stromal cells in colorectal cancer.

All artificial organ systems and medical devices that operate in direct contact with blood elicit activation of coagulation and platelets, and their long-term use often necessitates antithrombotic therapies that carry significant cost and bleeding risk. Thrombomodulin (TM) is a major endogenous inhibitor of blood coagulation localized on the endothelial cell surface. The overall objective of this research is to develop clinically durable synthetic materials by incorporating TM as a solid-supported film to actively and sustainably attenuate thrombus formation at the blood-contacting interface. During the course of this research, we developed site-specific approaches to covalently attach TM on the luminal surface of commercial vascular grafts using bioorthogonal chemistry that was compatible with ethylene oxide sterilization. Notably, we demonstrated the superior efficacy of TM to reduce platelet deposition compared with commercial heparin modified grafts using a non-human primate model of acute graft thrombosis. Finally, we optimized a novel reversible chemistry to rapidly and repeatedly regenerate immobilized TM, with the potential to significantly extend the lifetime of biologically active films.

Coagulation has and continues to be one of the most important elements in medicine. Issues from a lack of hemostasis range from poorer clinical outcomes to sudden death. The evolution of treatments for hemostasis have evolved from the use of Tamponade with direct pressure and bandages, the use of materials such as cobwebs and dust, the use of heat with hot oil or heated irons, to the use of suture, glues, plasmas, staplers, and electricity. This evolution has continued to bring about the prophylactic use of technology in an effort to prevent blood loss. This change from reactive treatments to proactive continue to be on a localized or superficial basis. One of the largest opportunities to proactively reduce blood loss in surgical patients is during the resection of solid organs such as the liver, kidney, and spleen. Few options have existed to help improve hemostasis short of the complete occlusion of blood supplying the tissue such as in the Pringle Maneuver. Recent studies have begun to show that practices such as this may have a significant detrimental effect on morbidity. It has been found that by applying radio frequency electrical energy in a particular way that large amounts of tissue can be pre-coagulated prior to resection. A series of animal and human clinical work has been completed to help evolve and confirm the method and the device that was created and refined during this effort. During the course of this work fifty-three patients were treated at four institutions on three continents. Average blood loss for liver resections performed with this pre-coagulation technique using the developed device in a multicenter control trail was 3.35 ml/cm2 as compared to 6.09 ml/cm2 (p < 0.05) for resections performed using standard surgical techniques alone. Additionally, the transection time necessary was also reduced from mean value of 27 minutes (2 -- 219 minutes) to 35 minutes (5 -- 65 minutes). Patients treated included those suffering from liver cirrhosis, fatty liver disease, and post chemotherapy fibrosis. From this work the use of pre-coagulation with methods and device developed was shown to be safe and effective for reducing the amount of blood loss and transection time during liver resections.

Project (M.S.)--Brigham Young University. Dept. of Statistics, 2007. / Includes bibliographical references (p. 60-61).

Diss. (sammanfattning) Uppsala : Univ., 2002. / Härtill 4 uppsatser.

Diss. (sammanfattning) Linköping : Univ., 2001. / Härtill 6 uppsatser.

Diss. (sammanfattning) Stockholm : Karol. inst., 2005. / Härtill 4 uppsatser.

Diss. (sammanfattning) Stockholm : Karolinska institutet, 2006. / Härtill 5 uppsatser.

Thesis (PhD.(Anatomy)--Faculty of Health Sciences)-University of Pretoria, 2007. / Includes bibliographical references.

Thesis (Ph. D.)--University of Missouri--Columbia, 1996. / Typescript. Vita. Includes bibliographical references (leaves : 138-152). Also available on the Internet.