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`PROCEEDINGS
`
`goth Annual Meeting of the
`American Association for Cancer Research
`April 10-14, 1999 e Philadelphia, PA
`VOLUME 40 et MARCH 1999
`
`CONTENTS
`(Arranged by Subject Category)
`
`:~B STRACTS
`pecial Multidisciplinary Sessions
`'l M ultidisciplinary and Translational Approaches to
`' Aerodigestive Cancers
`l
`\1 M ultidisciplinary and Translational Approaches to
`I Prostate Cancer
`l
`i arcinogenesis
`ONA Adducts, Protein Adducts, and Mutations
`carcinogen Metabolism
`Signaling Pathways in Tumor Promotion and
`Progression
`DNA Repair and Replication
`_I Hormones and Cancer
`Tobacco Carcinogenesis and Epidemiology
`Carcinogenesis in Experimental Models
`· ~netic Susceptibility in Experimental Models
`. i ~~asures of Human Cancer Susceptibility and Exposure
`.to Carcinogens
`, j Sgnal Transduction and Gene Expression in Ultraviolet
`\ ifand Chemical Carcinogenesis
`: 1 DUA Damage, Mutations, and Cellular Response
`'j ~markers of Premalignant and Malignant Lesions
`. , ..
`\ ! fJ!iA Adducts, Mutagenesis, and Repair
`Riactive Oxygen Species and Nitric Oxide
`! ~~.lecular and Enzymatic Determinants of
`lt arcmogenes1s
`.ic
`.
`.
`
`. ,, ~nd Tumor Biology
`
`t.
`
`} -,'
`
`' . .''el Therapies Targeting Angiogenesis
`· ' . stasis Models and Genetics
`,_
`'ii
`etics of Metastasis
`" f ptosis I
`'l',
`(';,ptosis II
`t·
`.
`h Factors and Receptors I
`" fr ular Signaling in Cancer Invasion and Metastasis
`;
`~ptosis Ill
`f
`(; ptosis JV
`, 'ulation of Angiogenic Factors
`ol of Apoptosis
`tosis V
`r-Host Interactions
`Factors and Receptors JI
`. l ·,:Tumor Interactions during Angiogenesis and
`t . erapeutic Approaches
`ion, Invasion, and Cellular Motility
`'
`-
`-·:; ',;genesis in Hormone Responsive Cancers
`. h Factors and Receptors Ill
`in Kinases in Tumor Progression/ Regression
`
`!"
`
`Page
`
`Cell and Tumor Biology (cont'd)
`
`Page
`
`Invasion: Its Regulation and Treatment
`Angiogenesis, Cell Signaling, and Gene
`Expression
`Cell Signaling in Tumor Progression
`Angiogenesis: Biology and Therapy
`Apoptosis VI: The Role of Mitochondria
`Metalloproteinases and Their Inhibitors
`Detection of Micrometastases
`Growth Factors and Receptors IV
`Interactions of Metastatic Cells with Their
`Environment: Genetic Implications
`Growth Factors and Receptors V
`
`Clinical Research
`
`Phase I Clinical Trials
`Phase I Trials of Novel Agents
`Lung Cancer I
`Head and Neck Cancer/Lung Cancer II
`Breast and Gynecologic Malignancies
`Prostate Cancer
`Bladder and Kidney Cancers
`Aerodigestive Cancers
`Gastrointestinal Cancer I: Colorectal and
`Hepatobiliary Cancers
`Gastrointestinal Cancer II: Hepatobiliary and Upper GI
`Tumors
`Molecular Pathogenesis and Tumor Progression
`Breast Cancer
`Gynecologic Malignancies
`Genitourinary Malignancies
`Melanoma/Sarcoma
`CNS and Pediatric Malignancies
`Lymphoma/Leukemia/Bone Marrow
`Transplantation
`Markers of Potential Clinical Utility
`
`Endocrinology/Preclinical and Clinical
`
`Endocrinology I
`Endocrinology II
`Retinoids and Their Signaling Pathways: Basic and
`Clinical Aspects
`Endocrinology Ill: Clinical and Translational Studies
`Endocrinology IV
`Hormones and Antihormones in the Regulation of Cell
`Growth and peath
`
`Epidemiology
`
`Cancer Epidemiology
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`518
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`561
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`713
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`730
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`232
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`316
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`337
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`343
`404
`491
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`524
`598
`603
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`718
`726
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`59
`157
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`308
`378
`613
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`636
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`39
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`202
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`408
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`44
`49
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`97
`153
`193
`246
`347
`350
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`410
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`501
`507
`607
`623
`644
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`740
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`65
`70
`103
`163
`168
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`West-Ward Exhibit 1086
`Geoerger Abs #3978
`Page 001
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`

`

`MDM2 positive specimens were also immunopositive for p53. Six p53 and MDM2
`double positive specimens exhibited no mutations in exons 5-8 by PCR-SSCP
`analysis. MDM2 overexpression was associated with nuclear anaplasia (p <
`0.05), previously linked with local recurrence of GCTs. The results indicate that
`MDM2 overexpression is a frequent abnormality in GCTs of bone and represents
`an alternative mechanism for inactivation of p53 in these tumors. The data
`suggest that MDM2 overexpression may contribute to the development of GCTs
`of bone.
`
`#3974 MAP and LAP expression in rhabdomyosarcomas of children and
`adults. Plaat, B.E.C., Van der Graaf, W.T .A., Mastik, M.F., Hollema, H., Hoekstra,
`H.J., Molenaar, W.M. Depts. of Pathology, Medical Oncology and Surgical On(cid:173)
`cology. University Hospital Groningen, The Netherlands.
`Introduction: Adult patients with rhabdomyosarcoma (RMS) have a worse
`prognosis than children. in which chemotherapy has improved clinical outcome.
`Pgp associated multidrug resistance in RMS is controversial. Therefore, the
`expression of Multidrug Resistance associated Protein (MRP) and the major vault
`protein LAP was examined in RMS of children and adults.
`Materials and Methods: On formalin fixed, paraffin embedded RMS tissue of 13
`children (<18 yrs of age) and 10 adults, the expression of MAP (MRPr1 ab) and
`LAP (Transduction Lab) were assessed immunohistologically. Percentage of
`stained cells and staining intensity were estimated and combined in a histopath(cid:173)
`score. Tumors <10% stained cells were considered negative. For statistics the
`Mann-Whitney U test and the Spearman correlation test were used.
`Results: LAP positive RMS were found in 8/13 children and 7/10 adults. MAP
`positive RMS were found in 4/13 children and 2/10 adults. LAP expression in
`adult RMS (mean: 40% pos. cells) was higher as compared to childhood RMS
`(mean: 30%). The MAP expression was lower in adult RMS (mean: 3%) as
`compared to childhood RMS (mean 13%). The differences were not statistically
`significant. Both LAP and MAP expression did not correlate with age.
`Conclusion: MAP and LAP are not clearly involved in the reported chemother(cid:173)
`apeutic resistance of adult RMS, since their expression is not associated with age
`and does not clearly differ between pediatric and adult RMS. Supported by the
`Dutch Cancer Society--Orant 95-1085.
`
`CLINICAL RESEARCH 16: CNS and Pediatric
`Malignancies
`
`#3975 Blocking G-protein function accounts for farnesylation inhibitor(cid:173)
`induced apoptosis in medulloblastoma cell lines. Wang, W. and Macaulay,
`R.J.B. University of Saskatchewan, Saskatoon, SK, Canada S7N 5E5.
`We have previously shown that blocking isoprenylation with lovastatin, an
`inhibitor of HMG-CoA reductase, or manumycin A. an inhibitor of protein fame(cid:173)
`syltransferase (FTase), inhibits medulloblastoma (MB) proliferation and induces
`apoptosis in vitro. We are investigating which of many intracellular famesylated
`proteins may be involved in these phenomena. Since ras and other G-proteins are
`famesylated, we used mycophenolic acid to deplete intracellular GTP, and thus
`block G-protein activation of downstream signalling cascades in vitro. We dem(cid:173)
`onstrate that blocking of G-protein function inhibits MB proliferation and induces
`apoptosis in a dose and time dependent manner. About 40 hours treatment with
`400 i-tM of mycophenolic acid induced the appearance of apoptotic DNA ladder(cid:173)
`ing in all tested cell lines (DAOY, UW228, 0283 Med and 0341 Med), while a
`higher concentration (700 µ.M) only needed 24 hours. Since !ovastatin has been
`shown to downregulate ras gene expression in yeast, we assessed this possibility
`in MB cell lines. The expression levels of total ras, and the K-. N- and H-ras genes
`were evaluated using RT-PCR with their respective specific primers. Following
`exposure to lovastatin and manumycin A in concentrations sufficient to induce
`apoptos1s, expression of the ras genes showed no significant change. Therefore.
`our results suggest that interference with G-protein function by blocking isopre(cid:173)
`nylation. rather than down-regulation of ras gene expression, contributes to
`lovastatin- and manumycin A-induced MB apoptosis. (Supported by the Brain
`Tumour Foundation of Canada).
`
`#3976
`1 H NMR of primary brain tumor extracts could allow the discrim(cid:173)
`ination between high-grade and low-grade gliomas. Sabatier, J., Gilard, V.,
`Malet-Martino, M .• Martino. R., Berry. I., and Tremoulet, M. IMRCP Lab., Univer·
`·
`site Paul Sabatier and CHU Purpan, Toulouse, France.
`Several choline-containing compounds (choline (Cho), phosphocholine (PC),
`glycerophosphochotine (GPC)) contribute to the <<choline>> peak observed in
`vivo at 3.2 ppm. whose significance is not well understood. In 1 H NMR spectra of
`glioma extracts. the resonances of the 3 compounds are separated with GPC at
`3.23 ppm, PC at 3.22 ppm and Cho at 3.21 ppm. Open surgery (6) or stereotactic
`biopsy (21) was conducted on 27 patients with brain tumors (20 high-grade and
`7 low-grade gtiomas). Tumor samples (6.0-98.5 mg) were extracted with perchlo(cid:173)
`ric acid. The absolute concentrations of choline-containing compounds (GPC +
`PC .,. Cho) in high-grade and low-grade gliomas were significantly different:
`0. 73 :!: 0.37 vs 0.36 :: 0.34 µmole/g (p < 0.025). The relative contributions of each
`
`CLINICAL RESEARCH 15
`
`of the choline-containing compound to the total choline signal were also statis(cid:173)
`tically different (except for Cho). For high-grade and low-grade gliomas, the
`percentages were: GPC, 30 :!: 11 % vs 68 :!: 13% (p < 0.0005), PC, 58 =. 14 % vs
`25 :: 8% (p < 0.0005), and Cho, 12 =. 9% vs 7 :!: 6% (p > 0.05). These data were
`corroborated by results obtained in 2 patients from samples of tissue from
`different regions (surface, tumor, core) of high-grade gliomas. Only the tumor
`tissue had a high-grade profile; the surrounding tissue had a profile of normal
`brain tissue and the core (necrosis) contained very low levels of metabolites. The
`differences in the concentration and the repartition of choline-containing com(cid:173)
`pounds seem to be a marker of high-grade gliomas. They could also help to
`discriminate between high· and low-grade gliomas in some difficult cases spe(cid:173)
`cially when there is histological uncertainty between anaplastic astrocytomas and
`low-grade oligodendrogliomas. (Supported by ARC, grant 6635).
`
`#3977
`In vitro evaluation of rapamycin sensitivity and synergy with cis(cid:173)
`platin and camptothecin in medulloblastoma. Kerr, Karol H., Sutton, Leslie N.
`and Phillips, Peter C. Division of Oncology, Children's Hospital of Philadelphia,
`Philadelphia, PA 19104.
`Medulloblastoma is the most common malignant Pediatric brain tumor. Al·
`though recent advances have been made in clinical trials with combination
`chemotherapy and radiation therapy, 20-40% of children will die secondary to
`relapsed disease and lack of adequate salvage therapy. Rapamycin (RAP). a
`immune-suppressant, is an agent with significant therapeutic potential for em·
`bryonal solid tumors. Seven medulloblastoma cell lines and one glioma cell line
`were tested for RAP sensitivity in an in vitro cellular proliferation assay. Cells
`demonstrated either exquisite RAP sensitivity (IC50 < 5 ng/mQ or high resistance
`(IC 50 > 800 ng/ml). A RAP sensitive cell line, DAOY, and a RAP resistant cell line,
`0283, were tested in drug synergy experiments. Rapamycin's cytotoxic effects
`were assayed with cisplatin (COOP) or camptothecin (CPT) alone and in combi·
`nation with RAP. RAP demonstrated a significant additive effect for both CDDP
`and CPT in the sensitive cell line only. Preliminary molecular studies identified
`potential signal transduction intermediaries involved in the RAP-dependent path·
`way in medulloblastoma. These finding suggest that raparnycin may be an out(cid:173)
`standing candidate for chemotheraputic clinical trials in medulloblastoma either
`alone or in combination with other agents. In addition, rapamycin represents a
`valuable probe to elucidate the tyrosine kinase • receptor linked pathways of
`signal transduction in medulloblastoma.
`
`#3978 Rapamycin analog CCI 779 inhibits growth of human medulloblas(cid:173)
`toma xenografts. Geoerger, B., Kerr, K., Janss. A.J., Sutton, LN. and Phillips,
`P.C. Children's Hospital of Philadelphia, Philadelphia, PA 19104.
`Rapamycin and other immunophilins with demonstrable antitumor activity have
`specific advantages for brain tumor therapy: relative lipophilicity; unique mecha(cid:173)
`nisms of cytotoxicity; absence of cross-resistance to other effective drugs; and
`minimal systemic toxicity. To evaluate the role of CCI 779, a rapamycin analog, in
`the treatment of medulloblastoma and other embryonal nervous system tumors,
`we examined in vitro cytotoxicity of CCI 779 in human medulloblastoma, neuro(cid:173)
`blastoma, and glioblastoma cell lines and in vivo activity of CCI 779 in athymic
`nude mice bearing DAOY medulloblastoma subcutaneous flank xenografts. In
`vitro cytotoxicity studies demonstrated 1050 s 1 O ng/ml in 4fi medulloblastoma
`and 1 /2 neuroblastoma cell lines. By contrast, a glioblastoma cell line (U251) was
`highly resistant (1050 > 1000 ng/ml). To evaluate in vivo cytotoxicity in athymic
`mice bearing DAOY medulloblastoma flank xenografts, CCI 779 was adminis(cid:173)
`tered i.p., daily x 5 for 1 or 2 weeks. Tumor volumes were measured serially and
`growth delay endpoints were defined as the post-treatment interval at which
`tumor volume increased by 5-fold. CCI 779 1-or 2-week treatments yielded
`significant tumor growth delays; i.e., the time to 5x initial tumor volume increased
`by 160% and 240%, respectively, compared to controls. Prolonged growth delay
`(> 50 days) was observed in 20% of the 2-week group bu1 not in the 1-week
`group. Retreatment of large tumors with CCI 779 restored growth inhibition but
`did not yield tumor regression. Our results indicate that prolonged treatment with
`CCI 779 causes significant growth delay in DAOY medulloblastoma and suggests
`that this class of immunophillns has major cytotoxic activity in neuroectodermal
`tumors.
`
`#3979
`lntratumoral immunotoxin treatment of human malignant brain
`tumors in nude animal models. Engebraaten, 0., Hjortland, G.-0., Juell, S .•
`Fodstad, 0. Department of Tumour Biology, Institute for Cancer Research, The
`Norwegian Radium Hospital, Oslo, Norway.
`Treatment of malignant brain tumors remains a clinical challenge. New treat·
`men! modalities have been introduced, and among these are infusion of immu(cid:173)
`notoxin molecules. recognizing specific cell surface epitopes on the tumor cells.
`We have compared the efficacy of two immunotoxins, Tfn-CRM107 and 425.3-PE
`targeted to the transferrin receptor and EGF receptor. respectively, in the treat(cid:173)
`ment of subcutaneous and intracranial glioma models in nude animals. lntratu(cid:173)
`moral administration of 1 µg Tfn-CRM107 into subcutaneously growing U87Mg
`cells efficiently inhibited tumor growth, whereas injection of 1 µ.g 425.3-PE were
`moderately effective. Treatment of intracerebral U87Mg tumors with Tfn-CRM107
`proved ineffective, as doses above 20 ng/animal were lethal lo the tumor bearing
`nude rats. In contrast, intratumoral administration of 425.3-PE delayed the onset
`
`Proceedings of the American Association for Cancer Research • Volume 40 o Marc.h 1999
`
`603
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`West-Ward Exhibit 1086
`Geoerger Abs #3978
`Page 002
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