First heterotransplantation of a human carcinoid tumor into nude mice

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125 express neuroendocrme cell markers including L-dopa-decarboxylase (DDC), creatine kinase-BB &o-enzyme (CK-BB), bombesin-like im- munoreactivity (BLI), neuron specific enolase (NSE), and neurosecre- tory type dense core granules (DGCs), compared to the parental cell line. All the lines stained positive for epithelial markers such as CAM5.2. LDH isoenzyme and chromosome analyses confirmed the human origin of all the cell lines. Therefore, it appears that cell line NCI-H69 contains stem cell subpopulation capable of generating cells of both small and non-small cell like phenotypes. Vasopressin elevates cytosolic calcium in small cell lung cancer cells Hong M, Moody TW. Department of Biochemistry and Molecular Bioiogy,George Washington University SchoolofMedicineandHealth Saences. 2300 Eye St. N. W., Washmgton. DC 20037. Peptides 1991;12:- 1315-9. The ability of vasopressin to elevate cytosolic CaZ* in small cell lung cancer (SCLC) cells was mvestlgated. Ten nanomolar vasopressm elevated the cytosolic Ca” in 6 of 8 SCLC cell lines that were loaded with Fura- AM. Using SCLC cell line NCI-H345, the effect of vasopressin was dose dependent, being maximal at 100 nM, where the cytosolic Ca” was elevated from 150 to 210 nM. Because addition of 1 mM EGTA had no effect on the vasopressin response, vasopressin released Ca*’ from intracellular pools. Also, oxytocin weakly elevated the cytosolic Ca”. The response to vasopressin was strongly blocked by [(B-mercapto-B,R-cyclopentamethylenepropionicacid)‘ ,O- MeTyri,Arg*]vasopressin and weakly blocked by [(O-mercapto-B.B- cyclopentamethylene proplow acid)‘0MeTyr’ ,Om’]vasotocin. These data suggest that V, vasopressm receptors are present on SCLC cells. Secondary screening system for preclinical testing of human lung cancer therapies Mulvin DW, Howard RB, Mitchell DH, Noker PE, Kmse CA, Chu HD et al. Division of Thoracic Surgery, Mount Sinai Hospital, 600 Univer- sity Ave., Toronto, Ont. M5G 1X.5. J Nat1 Cancer Inst 1992;84:31-7. The National Cancer Institute has instituted a primary screening system for testing new agents against cultured cancer cell lines. The purpose of this study was to determine the feasibility of using a nude rat orthotopic (organ-specific) human lung cancer model system as an in viva secondary screen for general evaluation of new anticancer agents and therapies active against lung cancer. To make this determination, we tested whether this system allows measurement of uptake and tumoricidal activity of anticancer therapies. Tumor-bearing lungs from 53 Rowett nude rats with orthotopically implanted human large-cell undifferenuated lungcarcmoma (NCI-H460) were perfused ex vwo for 1 hour with or without each of two anticancer modalities. Lungs were perfused wth blood-free perfusate alone (untreated control), perfusate with 100 pg/mL doxorubicin (treated positive control), or perfusate wth lymphokine-activated killer cells plus human recombinant inter- leukm-2 (LAK/rIL-2). Weight gain during perfusion was the criterion used to quantitate lung Injury. Treatment efficacy was measured by clonogenic assay after enzymatic disaggregation of the perfused tu- mors. Doxorubicin levels in the tumor and in the unmvolved lung were measured by high- performance hqmdchromatography. Both treatment groups showed only slight mcreascs in lung weight compared with that m the untreated control group, suggesting good lung tolerance of the procedure. Lung and tumor levels of doxorubicin were 320 It 21 ng/mg of tissue and 32 f 5 ng/mg of nssue (means f SE), respectively. Clonogenic assay demonstrated a fivefold to IO-fold reduction in the surviving fractxon of tumor cells with doxorubicin but no change with LAK/rIL-2. We conclude that thrs system allows demonstration of tumoruptakeandtumoric&lactivityofdoxorubicin hutthat itdoesnot demonstrate LAK/rIL-2 cytotoxiclty. The system may be useful for general secondary preclinical assessment of experimental lung cancer therapies. Further testing is needed before general use could be consid- ered. The ability of the system to allow detection of a dose-response relauonship as well as synergism m combined therapies is under mvestigation. Tumor-cell killing by MAbs against focosyl GMl, a ganglioside antigen associated with small-cell lung carcinoma Brezicka F-T, Holmgren J. Kalies I, Lindholm L. Department of Medical Microbiology and Immunology. Guldhedsgatan 10. S-413 46 Goteborg. Int J Cancer 1991;49:91 l-8. Monoclonal antibodies (MAbs) reactive with the ganglioside fucosyl GM1 (Fuc-GMI), an antigen associated with small-cell carcinoma of the lung (SCLC), were tested for their ability to mediate tumor-cell killing in vitro in conjunction with humoral and cellulareffectors and to inhibit tumor engraftment in nude mice in vivo. MAbs F12 and F15, both IgG(3k), induced human complement-mediated cytoloysis of 3 Fuc-GMI-expressing tumor cell lines: one rat hepatoma cell line, H4- II-E, and 2 human SCLC cell lines, NC1 H69 and NC1 H128. F12 and F15 also induced ADCC of these cell lines in the presence of either murine or human effector cells. Addition of sub-cytolytic amounts of fresh human serum as complement source resulted m enhanced ADCC induced by MAh F12 (IgG,). Also a Fuc-GMI-reactive MAb of IgM isotypz, F9, was able to induce such complement-aided ADCC (CADCC). F12 and F15 both proved to effectively inhibit engraftment of H4-II-E tumors in nude mice. A single dose of a modest amount (40 pg) of MAb conferred 65 to 100% protection against development of tumors. Our results demonsuate that Fuc-GM1 can act as a target antigen on tumor cells for specific immunotherapy m vitro and in a mouse model in viva. Complement and murine and human mononuclear effector cells were effective mediators of tumor cytolysis in vitro in the presence of mwne Fuc-GMI-reactive MAbs. Our results also suggest that humoral and cellular effecters may co-operate in specdic tumoricidal reacuons and that these may be induced by antibodies of both IgG and IgM tsotypes. Antiproliferative effects of the Ca*‘/calmodulin antagonist B&59-35 and the Cal*-channel blocker Verapamil on human lung cancer cell lines Schuller HM, Orloff M, Reznik GK. Experimental Oncology Labora- tory. Deportment of Pathobiology. College of Vetinary Medicine, Universily of Tennessee, Knoxville, TN 37901-1071. Carcinogenesis 1991;12:2301-3. We have recently demonstrated that the dihydropyridine- derivative 8859-35 has a selective chemotherapeutic effect on experimentally Induced neuroendocrine lung tumors in hamsters. These tumors re- sembled human atypical lung carcinoids morphologically and ex- pressed mammalian bombesin, calcitonin and neuron-specific enolase. In the hamster model, 8859-35 had no antiproliferative effect on pulmonary adenomas of Clara cell origin. In this study, we have tested the anuproliferative effects of 8859-35 and of the Ca”‘-channel blocker Verapamil in vitro on three human lung cancer cell hnes. The neuroen- docrine cell line NCI-H727 is derived from a lung carcinoid and expresses mammalian bombesin and calcitonin. Two non-neuroendo- wine cell lines are derived from peripheral pulmonary adenccarcino- ma& with line NCI-H322 expressing features of Clara cells while line NCI-H358 expresses features of alveolar type II cells. B859-35 was a potenl antiproliferative agent in the neumendocrine line NCI-H727 at concentrations as low as 0.001 pM, while it inhibited cell proliferation in the two other cell lines at concentrations of 100 nM and above. Verapamil inhibited cell proliferation in the neuroendocrine line NCI- H727 at concentrations of 1 nM and above. First heterobansplantation of a human carcinoid tumor into nude mice ‘“Janem YJ, Launay J-M. Debons-Guillemm M-C, Lasneret J, Roucay-. rol A-M, Lesser J et al. Service de Biochimie. Hopital Sainr-Louis, I Avenue Claude Vellefotu. 75475 Paris Cedex 10. Cancer 1991:68:893. 902. The first successful heterotransplantation of a human carcinoid rumorintonudemiceisreported.CSH,avoluminoushepaticmetastasis of a primary bronchial carcinoid tumor (CSB) was resected and trans- planted into three irradiated nude (Swiss-w/w) mice both by subcuta- neous (SC) and intramuscular (IM) routes: the success rate was five of

Transcript of First heterotransplantation of a human carcinoid tumor into nude mice

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express neuroendocrme cell markers including L-dopa-decarboxylase (DDC), creatine kinase-BB &o-enzyme (CK-BB), bombesin-like im- munoreactivity (BLI), neuron specific enolase (NSE), and neurosecre- tory type dense core granules (DGCs), compared to the parental cell line. All the lines stained positive for epithelial markers such as CAM5.2. LDH isoenzyme and chromosome analyses confirmed the human origin of all the cell lines. Therefore, it appears that cell line NCI-H69 contains stem cell subpopulation capable of generating cells of both small and non-small cell like phenotypes.

Vasopressin elevates cytosolic calcium in small cell lung cancer cells Hong M, Moody TW. Department of Biochemistry and Molecular Bioiogy,George Washington University SchoolofMedicineandHealth

Saences. 2300 Eye St. N. W., Washmgton. DC 20037. Peptides 1991;12:- 1315-9.

The ability of vasopressin to elevate cytosolic CaZ* in small cell lung cancer (SCLC) cells was mvestlgated. Ten nanomolar vasopressm elevated the cytosolic Ca” in 6 of 8 SCLC cell lines that were loaded with Fura- AM. Using SCLC cell line NCI-H345, the effect of vasopressin was dose dependent, being maximal at 100 nM, where the cytosolic Ca” was elevated from 150 to 210 nM. Because addition of 1 mM EGTA had no effect on the vasopressin response, vasopressin released Ca*’ from intracellular pools. Also, oxytocin weakly elevated the cytosolic Ca”. The response to vasopressin was strongly blocked by [(B-mercapto-B,R-cyclopentamethylenepropionicacid)‘,O- MeTyri,Arg*]vasopressin and weakly blocked by [(O-mercapto-B.B- cyclopentamethylene proplow acid)‘0MeTyr’,Om’]vasotocin. These data suggest that V, vasopressm receptors are present on SCLC cells.

Secondary screening system for preclinical testing of human lung cancer therapies Mulvin DW, Howard RB, Mitchell DH, Noker PE, Kmse CA, Chu HD et al. Division of Thoracic Surgery, Mount Sinai Hospital, 600 Univer- sity Ave., Toronto, Ont. M5G 1X.5. J Nat1 Cancer Inst 1992;84:31-7.

The National Cancer Institute has instituted a primary screening system for testing new agents against cultured cancer cell lines. The purpose of this study was to determine the feasibility of using a nude rat orthotopic (organ-specific) human lung cancer model system as an in viva secondary screen for general evaluation of new anticancer agents and therapies active against lung cancer. To make this determination, we tested whether this system allows measurement of uptake and tumoricidal activity of anticancer therapies. Tumor-bearing lungs from 53 Rowett nude rats with orthotopically implanted human large-cell undifferenuated lungcarcmoma (NCI-H460) were perfused ex vwo for 1 hour with or without each of two anticancer modalities. Lungs were perfused wth blood-free perfusate alone (untreated control), perfusate with 100 pg/mL doxorubicin (treated positive control), or perfusate wth lymphokine-activated killer cells plus human recombinant inter- leukm-2 (LAK/rIL-2). Weight gain during perfusion was the criterion used to quantitate lung Injury. Treatment efficacy was measured by clonogenic assay after enzymatic disaggregation of the perfused tu- mors. Doxorubicin levels in the tumor and in the unmvolved lung were measured by high- performance hqmdchromatography. Both treatment groups showed only slight mcreascs in lung weight compared with that m the untreated control group, suggesting good lung tolerance of the procedure. Lung and tumor levels of doxorubicin were 320 It 21 ng/mg of tissue and 32 f 5 ng/mg of nssue (means f SE), respectively. Clonogenic assay demonstrated a fivefold to IO-fold reduction in the surviving fractxon of tumor cells with doxorubicin but no change with LAK/rIL-2. We conclude that thrs system allows demonstration of tumoruptakeandtumoric&lactivityofdoxorubicin hutthat itdoesnot demonstrate LAK/rIL-2 cytotoxiclty. The system may be useful for general secondary preclinical assessment of experimental lung cancer therapies. Further testing is needed before general use could be consid- ered. The ability of the system to allow detection of a dose-response relauonship as well as synergism m combined therapies is under mvestigation.

Tumor-cell killing by MAbs against focosyl GMl, a ganglioside antigen associated with small-cell lung carcinoma Brezicka F-T, Holmgren J. Kalies I, Lindholm L. Department of Medical Microbiology and Immunology. Guldhedsgatan 10. S-413 46 Goteborg. Int J Cancer 1991;49:91 l-8.

Monoclonal antibodies (MAbs) reactive with the ganglioside fucosyl GM1 (Fuc-GMI), an antigen associated with small-cell carcinoma of the lung (SCLC), were tested for their ability to mediate tumor-cell killing in vitro in conjunction with humoral and cellulareffectors and to inhibit tumor engraftment in nude mice in vivo. MAbs F12 and F15, both IgG(3k), induced human complement-mediated cytoloysis of 3 Fuc-GMI-expressing tumor cell lines: one rat hepatoma cell line, H4- II-E, and 2 human SCLC cell lines, NC1 H69 and NC1 H128. F12 and F15 also induced ADCC of these cell lines in the presence of either murine or human effector cells. Addition of sub-cytolytic amounts of fresh human serum as complement source resulted m enhanced ADCC induced by MAh F12 (IgG,). Also a Fuc-GMI-reactive MAb of IgM isotypz, F9, was able to induce such complement-aided ADCC (CADCC). F12 and F15 both proved to effectively inhibit engraftment of H4-II-E tumors in nude mice. A single dose of a modest amount (40 pg) of MAb conferred 65 to 100% protection against development of tumors. Our results demonsuate that Fuc-GM1 can act as a target antigen on tumor cells for specific immunotherapy m vitro and in a mouse model in viva. Complement and murine and human mononuclear effector cells were effective mediators of tumor cytolysis in vitro in the presence of mwne Fuc-GMI-reactive MAbs. Our results also suggest that humoral and cellular effecters may co-operate in specdic tumoricidal reacuons and that these may be induced by antibodies of both IgG and IgM tsotypes.

Antiproliferative effects of the Ca*‘/calmodulin antagonist B&59-35 and the Cal*-channel blocker Verapamil on human lung cancer cell lines Schuller HM, Orloff M, Reznik GK. Experimental Oncology Labora- tory. Deportment of Pathobiology. College of Vetinary Medicine, Universily of Tennessee, Knoxville, TN 37901-1071. Carcinogenesis 1991;12:2301-3.

We have recently demonstrated that the dihydropyridine- derivative 8859-35 has a selective chemotherapeutic effect on experimentally Induced neuroendocrine lung tumors in hamsters. These tumors re- sembled human atypical lung carcinoids morphologically and ex- pressed mammalian bombesin, calcitonin and neuron-specific enolase. In the hamster model, 8859-35 had no antiproliferative effect on pulmonary adenomas of Clara cell origin. In this study, we have tested the anuproliferative effects of 8859-35 and of the Ca”‘-channel blocker Verapamil in vitro on three human lung cancer cell hnes. The neuroen- docrine cell line NCI-H727 is derived from a lung carcinoid and expresses mammalian bombesin and calcitonin. Two non-neuroendo- wine cell lines are derived from peripheral pulmonary adenccarcino- ma& with line NCI-H322 expressing features of Clara cells while line NCI-H358 expresses features of alveolar type II cells. B859-35 was a potenl antiproliferative agent in the neumendocrine line NCI-H727 at concentrations as low as 0.001 pM, while it inhibited cell proliferation in the two other cell lines at concentrations of 100 nM and above. Verapamil inhibited cell proliferation in the neuroendocrine line NCI- H727 at concentrations of 1 nM and above.

First heterobansplantation of a human carcinoid tumor into nude mice ‘“Janem YJ, Launay J-M. Debons-Guillemm M-C, Lasneret J, Roucay-. rol A-M, Lesser J et al. Service de Biochimie. Hopital Sainr-Louis, I Avenue Claude Vellefotu. 75475 Paris Cedex 10. Cancer 1991:68:893. 902.

The first successful heterotransplantation of a human carcinoid rumorintonudemiceisreported.CSH,avoluminoushepaticmetastasis of a primary bronchial carcinoid tumor (CSB) was resected and trans- planted into three irradiated nude (Swiss-w/w) mice both by subcuta- neous (SC) and intramuscular (IM) routes: the success rate was five of

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six. Heterotmnsplanted tumors took 4 to 5 months to appear in the mice and 1 month to attain a width of 0.5 cm. Both human and mouse tumors (named CSH-SC and CSH-IM) were studied by light and electron microscopy. They were Grimelius-positive, neuron-specific enolase- positive, and born&in-negative by immunocytochemistry. Further- more,CSH-SCcellspresentedcharacteristic @car-shaped.rod-shaped, or tadpole-shaped) neurosecretory granules: Although CSB and CSH were slightly serotonin positive by immunocytochemistry, only a few serotonin-positive cells were found in CSH-SC and none in CSH-IM, suggesting partial loss of differentiation or an increase in serotonin catabolism during transplantation.

Detection of suspected primary lung cancer by scintigraphy wth indium-111-anti-carcinoembryonic antigen mono&ma1 antibodies (type FlwC5) BiggL A, Buccheri G, Ferrigno D, Viglietti A, Farinelli MC, Comino A et al. Servizio di Medicina Nucleare, Ospedale S. Croce, via Coppino 25.12100 Cuneo. J Nucl Med 1991;32:2064-8.

Immunoscintigraphy with “‘In-labeled anti-CEA-Mab (FW23C51) was carried out in 66 patients strongly suspected for a primary lung cancer and in 8 control patients suffering from different chest diseases. A sensitivity of 0.90, a specificity of 0.45 and an accuracy of 0.85 were calculated. False-negative results were mainly obtained in patients in whom the size of the lesion was below 2 cm and the tumor was cenually located. All patients affected by small-cell carcinoma were correctly identified. In 89% of the patients, a positive immunoscintiscan was associated with the presence of the antigen in the tumor. False-positive results were observed in control patients suffering from different chest diseases due to the nonspecific uptake of the tracer. The tumor defini- tion was generally better after 120 hr than at an earlier time after injection due to the reduction of background activity. SPECT imaging defined the tumor better in each patient but did not reveal any tumor not seen on planar studies.

Genetic susceptibility to squamous cell carcinoma of the lung in relation to cigarette smoking dose Nakachi K, Imai K, Hayashi S-I, Watanabe J, Kawajiri K. Deparfment of Epidemiology, Sailam0 Cancer Center Research Ins&u@, 818 &mum, Inn-machi, Saimmo 362. Cancer Res 1991;Sl:S177-80.

Cytochrome P4SOIAl is responsible for the metabolic activation of benzo(a)pyrene in cigarette smoke; an association of lung cancer with DNA polymorphisms of P450IAl gene was shown in our previous study. In this paper, we investigated the interindividual difference of genetically determined susceptibility to squamouscellcarcinomaofthe lung in relation to cigarette smoking dose. We first compared the total amounts of cigarettes consumed over dte lifetime of patients and showed that the patients with a “susceptible” P4501Al gene genotype contracted carcinoma after fewer cigarettes [mean f SD, 31.3 f 12.8 x l(r cigarettes (n = 12)] than those with other genotypes I42.5 i 18.2 x lO’(n = 33)], with a statistical significanceof P-z 0.05. Next, we carried out a case-control study to estimate tie odds ratios of susceptible to nonsusceptible individuals in relation to the cumulated cigarette dose. We thus showed that the individuals with the susceptible genotype were at remarkably high risk with an odds ratio of 7.31 (95% confidence interval, 2.13 1025.12) at a low dose level ofcigarette smoking and that the difference in susceptibility between genotypes was reduced at high dose levels.

Production of neuromedin Band neuromedin B gene expression in human lung tumor cell lines Cardona C. Rabbitts PH. Spindel ER, Ghatei MA, Bleehen NM et al. Medical Research Council, Clinical Oncology and Radiotherapeulics Unir, MRC Cenrre, Hills Road, Cambridge CB2 2QH. Cancer Res 1991:51:5205-11.

Gastrin-releasing peptide (GRP), a mammalian bombesin-like pep tide, has been shown to be an important autocrine growth factor for small cell lung cancer (SCLC). However, not all SCLC cell lines express the GRP gene or respond mitogenically to GRP stimulation,

suggesting the existence of other autocrine pathways in this tumor. Neuromedin B (NMB), the mammalian counterpart of amphibian mnatensin, has been shown to be a mitogen for SCLC cell lines in vitro. TO determine whether NMB is a potential autcxrine growth factor for lung tumors, NMB gene expression, peptide synthesis, and secretion have been investigated in a panel of SCLC and non-SCLC (NSCLC) cell lines. Northern blot analysis and enzymatic amplification from mRNA by polymerase chain reaction showed that the NMB gene was expressed in all SCLC and NSCLC cell lines examined. In contrast, the GRP gene was expressed in four of six classic SCLC cell lines but not in variant SCLC or NSCLC cell lines. lmmunoreactive NMB was detected by radioimmunoassay in the majority of classic SCLC, in one of three variant SCLC and in one of three NSCLC cell lines, and secreted NMB was detected in medium conditioned by a SCLC and a NSCLC cell line. The present study also demonstrated the presence of lmmunoreactive GRP in the absence of detectable GRP gene expres- sion. The antiserum used in the GRP radioimmunoassay failed to cross- react with NMB but showed some cross-reactivity with amphibian phyllolitorin raising the possibility that certain SCLC cell lines may produce a phyllolitorin-like peptide.

Different energy metabolism in two human small cell lung cancer subpopulations examined by “P magnetic resonance spectroscopy and biochemical analysis in viva and in vitro Kristjansen PEG, Spang-Thomsen M, Quistorff B. Deparmenf of Oncology, Finsen Insrime-Rigshospirler. Blegdamsvej 9, DK-2100 Copenhagen. Cancer Res 1991;51:5160-4.

Two human small cell lung cancer tumor lines, maintained as solid tumorxenograftsonnude miceandas in vitrocell cultures, were studied by in vivo ,‘P magnetic resonance spectroscopy and by biochemical analysis of extracts of solid tumors and cell cultures. Thd tumor lines CPH SCCL 54A and CPH SCCL 548 are subpopulations from the same tumor. In solid tumors (n = 125). the ATP/P(i) ratio was greater in 54A than in 548. This was due to a higher ATP level in 54A, whereas there was no difference in P(i), ADP, and AMP. A decrease in ATPiP during growth was caused by a decline in ATP, whereas P(i) remained unchanged. Small amounts of phosphocreatine were found in the xenografts and in tumor extracts, but not in the cell extracts; corre- spondingly, there was a low creatine kinase activity in solid tumors and no activity in the cell cultures. Thus, the phosphocreatine content of the solid tumors originated from the stroma. A difference in ATP content between 54A and 54B was also found in cell cultures; hence, the metabolic difference is an intrinsic quality of the malignant cells and is not caused by the host system.

Allelic loss on the short arm of chromosome 11 in non-small-cell lung cancer Ludwig CU, Raefle G, Dalquen P, Stulz P. Stahel R, Obrecht J-P. Section for Oncology, Department of Inrernal Medicine, University Hospiml, CH-1032 Bawl. Int J Cancer 1991;49:661-5.

Forty-eight samples of primary non-small lung cancer (NSCLC) and normal tissue from the same patients were analyzed forallelic deletions on chromosome 11~. five polymorphic loci were assessed to determine the incidence of IIp sequence deletions and to define hot-spots of deletions. Information was obtained from all patients in at least one locus. Our data show that the deletions observed were not ramdomly scattered over the short arm of chromosome II. Rather, 2 hot-spots of deletions were observed: one in the area of the genes for catalase and & FSH corresponding to band 11~13, the other close to the IGF-II locus corresponding to band IIplS. A high incidence of loss of hetrozygosity f&OH) was found with the probe for catalase (21/29), a locus flanking the centromeric region of the Wilms’ tumor locus. Most of the samples exhibiting LOH of one or more of the alleles analyzed remained heterozygous for at leastoneotherchromosome IIpallele.Furthermore, duplication of the intensity of th eremaining allele was rarely observed. Our results indicate that LOH on the short arm of chromosome II is a common event in NSCLC and that the chromosomal region containing the Wilms’ tumor locus is most commonly involved.