Prevention of Chemotherapy-Induced Anemia and Thrombocytopenia by Constant Administration of Stem Cell Factor

Purpose: Chemotherapy-induced apoptosis of immature hematopoietic cells is a major cause of anemia and thrombocytopenia in cancer patients. Although hematopoietic growth factors such as erythropoietin and colony-stimulating factors cannot prevent the occurrence of drug-induced myelosuppression, stem cell factor (SCF) has been previously shown to protect immature erythroid and megakaryocytic cells in vitro from drug-induced apoptosis. However, the effect of SCF in vivo as a single myeloprotective agent has never been elucidated. Experimental Design: The ability of SCF to prevent the occurrence of chemotherapy-induced anemia and thrombocytopenia was tested in a mouse model of cisplatin-induced myelosuppression. To highlight the importance of maintaining a continuous antiapoptotic signal in immature hematopoietic cells, we compared two treatment schedules: in the first schedule, SCF administration was interrupted during chemotherapy treatment and resumed thereafter, whereas in the second schedule, SCF was administered without interruption for 7 days, including the day of chemotherapy treatment. Results: The administration of SCF to cisplatin-treated mice could preserve bone marrow integrity, inhibit apoptosis of erythroid and megakaryocytic precursors, prevent chemotherapy-induced anemia, and rapidly restore normal platelet production. Treatment with SCF increased the frequency of Bcl-2/Bcl-XL\u2013 positive bone marrow erythroid cells and sustained Akt activation in megakaryocytes. Myeloprotection was observed only when SCF was administered concomitantly with cisplatin and kept constantly present during the days following chemotherapy treatment. Conclusions: SCF treatment can prevent the occurrence of chemotherapy-induced anemia and thrombocytopenia in mice, indicating a potential use of this cytokine in the supportive therapy of cancer patients

Tumoral calcinosis: Identification of a novel recessive mutation in fibroblast growth factor-23 (FGF-23)

Tumoral calcinosis (TC) is a rare genetic disorder characterized by periarticular cystic and solid tumorals calcifications. It is characterized by hyperphospatemia and an elevated serum of calcitriol concentration in every patients. The hyperphosphatemia results form an increase in capacity of renal tubular phosphate reabsorption. The identification of phosphotonin family hormones suggest that mutations of these molecules could be involved in the pathogenesis of TC. One of these molecules is represented by FGF-23. The TC phenotype is similar to that described in the FGF-23 knockout mice. In the present study we described a new FGF-23 mutation in a subject affected by TC.A Caucasian women (years 67) was examined for a history ofectopic calcification. Biochemical exams showed and hyperphosphathemia and hyperphosphaturia with normal value ofPTH and inappropriately normal level of 1–25 (OH)2 D3. The patient presented a big shoulder calcification and also a calcification of femoral artery. We expanded the family tree through detailed family histories, which importantly revealed that parents were consanguineous. Hystologically the mass was characterized by calcium deposition and granulomatous reaction around the mass. Genomic DNA was extracted from blood collected from the patient, her daughter and her grandchild by standard procedure. DNA was not available from her parents. All three FGF-23coding exons, as well as conserved splice sites, were amplified by standard PCR procedure. Nucleotide sequences were determined by direct sequencing with a DNA kit and an automated DNA sequencer (ABI PRISM 3100 - Perkiln-Elmer Corp). We discovered a new homozygous codon 41, His/Gln (CAC-CAA) substitution in exon 1 of FGF-23 gene in the affected patient. A heterozygous substitution was present in the daughter. No mutation were found in the two children. FGF-23 gene mutation was not found in the SNP database (www.ncbi.nih.gov/snp). In summary, a recessive mutation in FGF 23 causes TC. Understanding the functional significance and molecular physiology of this novel mutation will reveal critical information regarding the role of FGF-23 in states of normal and of disorder of phosphate homeostasis

Methodological models for in vitro amplification and maintenance of human articular chondrocytes from elderly patients

Articular cartilage defects, an exceedingly common problem closely correlated with advancing age, is characterized by lack of spontaneous resolution because of the limited regenerative capacity of adult articular chondrocytes. Medical and surgical therapies yield unsatisfactory short-lasting results. Recently, cultured autologous chondrocytes have been proposed as a source to promote repair of deep cartilage defects. Despite encouraging preliminary results, this approach is not yet routinely applicable in clinical practice, but for young patients. One critical points is the isolation and ex vivo expansion of large enough number of differentiated articular chondrocytes. In general, human articular chondrocytes grown in monolayer cultures tend to undergo dedifferentiation. This reversible process produces morphological changes by which cells acquire fibroblast-like features, loosing typical functional characteristics, such as the ability to synthesize type II collagen. The aim of this study was to isolate human articular chondrocytes from elderly patients and to carefully characterize their morphological, proliferative, and differentiative features. Cells were morphologically analyzed by optic and transmission electron microscopy (TEM). Production of periodic acid-schiff (PAS)-positive cellular products and of type II collagen mRNA was monitored at different cellular passages. Typical chondrocytic characteristics were also studied in a suspension culture system with cells encapsulated in alginate-polylysine-alginate (APA) membranes. Results showed that human articular chondrocytes can be expanded in monolayers for several passages, and then microencapsulated, retaining their morphological and functional characteristics. The results obtained could contribute to optimize expansion and redifferentiation sequences for applying cartilage tissue engineering in the elderly patients