Neuroblastoma is the most common extra-cranial solid tumor of infancy and childhood, and majority of patients die from the metastatic disease. Orthotopic xenograft mouse models are valuable tools for improving our understanding and control of neuroblastoma metastasis, because they readily represent genetic diversity and allow spontaneous metastasis. Intra-adrenal injection is commonly used for establishing the orthotopic animal models since human neuroblastoma frequently originates in the adrenal gland. However, it is unclear whether the metastatic potential of neuroblastoma can be reliably determined in adrenally-injected mice because their gland size is so small. In this study, we developed and characterized a fluorescent orthotopic xenograft animal model of NB69-derived human neuroblastoma. By comparing animals receiving adrenal injection and adrenal overlay, with the latter mimicking injection spillover, we found that the metastatic potential of neuroblastoma can be reliably determined in animal lungs. Furthermore, the lung metastasis can be genetically modulated in these animals. The results also show that the expression of Renilla green fluorescent protein (GFP) was exceptionally stable in NB69 cells, allowing rapid and sensitive detection of lung metastases at the macroscopic level. Additional features of our model include 100% tumor take, a 1-week tumor latency, resemblance to tumor behaviors in neuroblastoma patients, and the ability to monitor the expression of a gene of interest with GFP. This animal model of human neuroblastoma will be useful for studying genes involved in the metastatic process and for evaluating anti-metastasis agents in pre-clinical trials.

Nucleoside diphosphate kinase A (NDPK-A), encoded by the nm23-H1 gene, acts as a metastasis suppressor in certain human tumors such as breast carcinoma. However, evidence also points to NDPK-A functioning as a metastasis promoter in other human tumors including neuroblastoma. In fact, amplification and overexpression of nm23-H1 as well as S120G mutation of NDPK-A (NDPK-AS120G) have been detected in 14% to 30% of patients with advanced stages of neuroblastoma. To test whether NDPK-A promotes neuroblastoma metastasis, we established stable transfectants and an orthotopic xenograft animal model from the human neuroblastoma NB69 cell line. We demonstrate that overexpressed NDPK-A or NDPK-AS120G increased both incidence and colonization of neuroblastoma metastasis in animal lungs without significantly affecting primary tumor development. In vitro, these metastasis-associated NDPK-A aberrations abrogated retinoic acid-induced neuronal differentiation while increasing cloning efficiency, cell survival, and colony formation of NB69 derivatives. Furthermore, NDPK-AS120G reduced cell adhesion and increased cell migration. Compared with its wild-type, NDPK-AS120G appears more effective in promoting neuroblastoma metastasis. Our results provide the first evidence that NDPK-A behaves as a metastasis promoter at least in human neuroblastoma derived from NB69 cells. The findings not only suggest a prognostic value of NDPK-A in neuroblastoma patients but also caution NDPK-A-targeted treatment for patients with different tumor types.

Diseases inherited as simple recessive frequently reflect simple anomalies in the structure of enzymic proteins. Many such inherited diseases of humans have now been defined in biochemical terms, but relatively few have been described adequately in domestic animals. Collagen is reported to be involved in many diseases in man (PINNELL et al., 1972) and animals (O'HARA et al., 1970). FJ~LSTAD and HELLE (1974) reported an inherited disease in Dala sheep, characterized by a defect in the normal packing of collagen molecules into fibres. The clinical diagnostic feature of a collagenous tissue dysplasia disease is considered to be fragility of the skin, leading to the formation of papyraceous scars. This disease has many clinical characteristics in common with the Ehlers-Danlos syndrome of man (PEARSE, 1964), cutaneous asthenia of dogs and mink (HEGREBERG et al., 1969), and dermatosparaxis of calves (ANSAY et al., 1968). A diesease with a similar clinical appearance has now been recognized in Finnish crossbred sheep.The purpose of this paper is to present the gross pathology, histo-and cytopathology of the skin of clinically affected animals, and some blood parameters of ewes giving normal and ewes giving defective lambs. Material and MethodsBreeding histo y of affected flocks Farm 1. The flock was established at Lohja province early in 1977 from 9 Finnsheep ewes and one ram. No defective lambs were noticed at the first lambing. In 1978, a Suffolk ram ( S ) was brought to the flock and the ewes were sired by this ram. He produced 30 daughters; again no defective lambs $ere noticed. In 1979 the ewelambs (i. e. the 30 daughters) were sired by their brother ram (M), also a progeny of ram (S). Of the progeny born three lambs were defective. The ewes involved have never produced more than one defective lamb. This is the type of situation where recessive defects might be expected to arise. Year-by-year lambing performance of ewes has left defective offspring of both sexes U. S.