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The activity of camptothecin analogues is enhanced in histocultures of human tumors and human tumor xenografts by modulation of extracellular pH

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Abstract

Background

Most solid human tumors exist in an acidic microenvironment, due in part to inefficient vasculature and a higher intrinsic rate of glycolysis. This leads to a tumor-selective pH gradient, which can be exploited therapeutically with antitumor agents such as the camptothecins (CPTs). Previous work in this laboratory has shown that camptothecin activity is enhanced 40- to 60-fold in monolayer cell culture by reducing the extracellular pH to 6.8. Three-dimensional histoculture has been shown to be a technique that allows human tumor tissue to grow in an in vivo-like way with maintenance of tissue histology and function and drug sensitivity for long periods of time.

Purpose

In the current study, we utilized these features of histoculture to study new analogues of camptothecin that have superior pharmacological properties.

Methods

We evaluated six CPT analogues in histocultures of human brain, neuroblastoma, breast, colon, and prostate tumors. Fragments were exposed to 10,11-methylenedioxy-CPT (MDC), 7-chloromethyl-MDC, SN-38, topotecan (TPT), 9-amino-CPT, 10-amino-CPT, paclitaxel, 5-fluorouracil, 4-hydroperoxycyclophosphamide and doxorubicin, and antitumor activity was assessed. For in vivo tumor outgrowth studies, fragments were treated in parallel, implanted into nude mice, and monitored for development of tumors.

Results

Against 15 of 16 tumor xenografts and all primary tumor samples tested, all compounds were cytotoxic at pH 7.4 (IC50 range 13–921 μM). MDC, SN-38, TPT, and 9-amino-CPT achieved an average 5-fold increase in activity (range 3–14) at pH 6.8, while 7-chloromethyl-MDC was enhanced 8-fold (range 6–14). The most potentiated analogue was 10-amino-CPT at 27-fold (range 17–49). In contrast, the other agents were active against one or more tumor types but were not enhanced by acidic pH. Importantly, the toxicity of MDC in histoculture of D54 glioma xenografts strongly correlated with the outgrowth of treated fragments subsequently implanted in vivo.

Conclusion

Evaluation of anticancer drug activity in native-state histoculture supports the concept that pH modulation may be an important approach to improve the selectivity and antitumor effectiveness of camptothecin-based chemotherapy.

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References

  1. Adams DJ, Dewhirst MW, Flowers JL, Gamcsik MP, Colvin OM, Manikumar G, Wani MC, Wall ME (2000) Camptothecin analogues with enhanced antitumor activity at acidic pH. Cancer Chemother Pharmacol 46:263

    Article  CAS  PubMed  Google Scholar 

  2. Baibakov B, Frank GA, Sergeeva N, Novikova I, Youngkin T, Connors KM, Hoffman RM, Margolis LB (1991) In vivo growth patterns of human lung tumors in three-dimensional histoculture. In Vitro Cell Dev Biol 27A:897

    CAS  PubMed  Google Scholar 

  3. Burke TG, Mi Z (1993) Preferential binding of the carboxylate form of camptothecin by human serum albumin. Anal Biochem 212:285

    Article  CAS  PubMed  Google Scholar 

  4. Burke TG, Mishra AK, Wani MC, Wall ME (1993) Lipid bilayer partitioning and stability of camptothecin drugs. Biochemistry 32:5352

    CAS  PubMed  Google Scholar 

  5. Burke TG, Munshi CB, Mi Z, Jiang Y (1995) The important role of albumin in determining the relative human blood stabilities of the camptothecin anticancer drugs. J Pharm Sci 84:518

    CAS  PubMed  Google Scholar 

  6. Chang SG, Jung JC, Rho YS, Huh JS, Kim JI, Hoffman RM (1996) Efficacy of the platinum analog [Pt(cis-dach)(DPPE)-2NO3] on histocultured human patient bladder tumors and cancer cell lines. Anticancer Res 16:3423

    CAS  PubMed  Google Scholar 

  7. Cohen DP, Adams DJ, Flowers JL, Wall ME, Wani MC, Manikumar G, Colvin OM, Silber R (1999) Pre-clinical evaluation of SN-38 and novel camptothecin analogs against human chronic B-cell lymphocytic leukemia lymphocytes. Leuk Res 23:1061

    Article  CAS  PubMed  Google Scholar 

  8. Connor CE, Norris JD, Broadwater G, Willson TM, Gottardis MM, Dewhirst MW, McDonnell DP (2001) Circumventing tamoxifen resistance in breast cancers using antiestrogens that induce unique conformational changes in the estrogen receptor. Cancer Res 61:2917

    CAS  PubMed  Google Scholar 

  9. Driscoll TA, Flowers JL, Colvin OM (2002) Potentiation of camptothecin-based anti-neuroblastoma activity by selective tumor acidification. In: Proceedings of the 93rd Annual Meeting of the American Association of Cancer Research, San Francisco

  10. Engin K, Leeper DB, Cater JR, Thistlethwaite AJ, Tupchong L, McFarlane JD (1995) Extracellular pH distribution in human tumours. Int J Hyperthermia 11:211

    Google Scholar 

  11. Flowers JL, Hoffman RM, Manikumar G, Wani MC, Wall ME, Colvin OM, Adams DJ (2002) Activity of camptothecin analogues against human prostate cancer cells in monolayer and in 3-dimensional histoculture as a function of extracellular pH. In: Proceedings of the 93rd Annual Meeting of the American Association of Cancer Research, San Francisco

  12. Freeman AE, Hoffman RM (1986) In vivo-like growth of human tumors in vitro. Proc Natl Acad Sci U S A 83:2694

    PubMed  Google Scholar 

  13. Furukawa T, Kubota T, Hoffman RM (1995) Clinical applications of the histoculture drug response assay. Clin Cancer Res 1:305

    PubMed  Google Scholar 

  14. Gamcsik MP, Kasibhatla MS, Adams DJ, Flowers JL, Colvin OM, Manikumar G, Wani MC, Wall ME, Kohlhagen G, Pommier Y (2001) Dual role of glutathione in modulating camptothecin activity: depletion potentiates activity, but conjugation enhances the stability of the topoisomerase I-DNA cleavage complex. Mol Cancer Ther 1:11

    CAS  PubMed  Google Scholar 

  15. Geller J, Sionit LR, Connors K, Hoffman RM (1992) Measurement of androgen sensitivity in the human prostate in in vitro three-dimensional histoculture. Prostate 21:269

    CAS  PubMed  Google Scholar 

  16. Geller J, Sionit LR, Connors KM, Youngkin T, Hoffman RM (1993) Expression of prostate-specific antigen in human prostate specimens in in vitro three dimensional histoculture. In Vitro Cell Dev Biol 29A:523

    CAS  Google Scholar 

  17. Geller J, Sionit LR, Baird A, Kohls M, Connors KM, Hoffman RM (1994) In vivo and in vitro effects of androgen on fibroblast growth factor-2 concentrations in the human prostate. Prostate 25:206

    CAS  PubMed  Google Scholar 

  18. Geller J, Partido C, Sionit L, Youngkin T, Nachtsheim D, Espanol M, Tan Y, Hoffman R (1997) Comparison of androgen-independent growth and androgen-dependent growth in BPH and cancer tissue from the same radical prostatectomies in sponge-gel matrix histoculture. Prostate 31:250

    Article  CAS  PubMed  Google Scholar 

  19. Geller J, Sionit L, Partido C, Li L, Tan X, Youngkin T, Nachtsheim D, Hoffman RM (1998) Genistein inhibits the growth of human-patient BPH and prostate cancer in histoculture. Prostate 34:75

    Article  CAS  PubMed  Google Scholar 

  20. Gerweck LE, Seetharaman K (1996) Cellular pH gradient in tumor versus normal tissue: potential exploitation for the treatment of cancer. Cancer Res 56:1194

    CAS  PubMed  Google Scholar 

  21. Gerweck LE, Kozin SV, Stocks SJ (1999) The pH partition theory predicts the accumulation and toxicity of doxorubicin in normal and low-pH-adapted cells. Br J Cancer 79:838

    Google Scholar 

  22. Green SK, Frankel A, Kerbel RS (1999) Adhesion-dependent multicellular drug resistance. Anticancer Drug Des 14:153

    CAS  PubMed  Google Scholar 

  23. Guadagni F, Roselli M, Hoffman RM (1991) Maintenance of expression of tumor antigens in three-dimensional in vitro human tumor gel-supported histoculture. Anticancer Res 11:543

    CAS  PubMed  Google Scholar 

  24. Hoffman RM (1998) Three-dimensional sponge-gel matrix histoculture of human tumors: methods and applications, 1st edn. Academic Press, New York

  25. Hoffman RM, Connors KM, Meerson-Monosov AZ, Herrera H, Price JH (1989) A general native-state method for determination of proliferation capacity of human normal and tumor tissues in vitro. Proc Natl Acad Sci U S A 86:2013

    PubMed  Google Scholar 

  26. Hsiang YH, Hertzberg R, Hecht S, Liu LF (1985) Camptothecin induces protein-linked DNA breaks via mammalian DNA topoisomerase I. J Biol Chem 260:14873

    PubMed  Google Scholar 

  27. Hsiang YH, Lihou MG, Liu LF (1989) Arrest of replication forks by drug-stabilized topoisomerase I-DNA cleavable complexes as a mechanism of cell killing by camptothecin. Cancer Res 49:5077

    CAS  PubMed  Google Scholar 

  28. Jaxel C, Kohn KW, Wani MC, Wall ME, Pommier Y (1989) Structure-activity study of the actions of camptothecin derivatives on mammalian topoisomerase I: evidence for a specific receptor site and a relation to antitumor activity. Cancer Res 49:1465

    PubMed  Google Scholar 

  29. Kerbel RS, Rak J, Kobayashi H, Man MS, St Croix B, Graham CH (1994) Multicellular resistance: a new paradigm to explain aspects of acquired drug resistance of solid tumors. Cold Spring Harb Symp Quant Biol 59:661

    CAS  PubMed  Google Scholar 

  30. Kollmannsberger C, Mross K, Jakob A, Kanz L, Bokemeyer C (1999) Topotecan—a novel topoisomerase I inhibitor: pharmacology and clinical experience. Oncology 56:1

    CAS  PubMed  Google Scholar 

  31. Kubota T, Sasano N, Abe O, Nakao I, Kawamura E, Saito T, Endo M, Kimura K, Demura H, Sasano H, et al (1995) Potential of the histoculture drug-response assay to contribute to cancer patient survival. Clin Cancer Res 1:1537

    PubMed  Google Scholar 

  32. Luzzio MJ, Besterman JM, Emerson DL, Evans MG, Lackey K, Leitner PL, McIntyre G, Morton B, Myers PL, Peel M, et al (1995) Synthesis and antitumor activity of novel water soluble derivatives of camptothecin as specific inhibitors of topoisomerase I. J Med Chem 38:395

    CAS  PubMed  Google Scholar 

  33. Mi Z, Burke TG (1994) Differential interactions of camptothecin lactone and carboxylate forms with human blood components. Biochemistry 33:10325

    CAS  PubMed  Google Scholar 

  34. Olbina G, Miljkovic D, Hoffman RM, Geller J (1998) New sensitive discovery histoculture model for growth-inhibition studies in prostate cancer and BPH. Prostate 37:126

    Article  CAS  PubMed  Google Scholar 

  35. Perou CM, Sorlie T, Eisen MB, van de Rijn M, Jeffrey SS, Rees CA, Pollack JR, Ross DT, Johnsen H, Akslen LA, Fluge O, Pergamenschikov A, Williams C, Zhu SX, Lonning PE, Borresen-Dale AL, Brown PO, Botstein D (2000) Molecular portraits of human breast tumours. Nature 406:747

    CAS  PubMed  Google Scholar 

  36. Philippart P, Harper L, Chaboteaux C, Decaestecker C, Bronckart Y, Gordover L, Lesueur-Ginot L, Malonne H, Lavergne O, Bigg DC, da Costa PM, Kiss R (2000) Homocamptothecin, an E-ring-modified camptothecin, exerts more potent antiproliferative activity than other topoisomerase I inhibitors in human colon cancers obtained from surgery and maintained in vitro under histotypical culture conditions. Clin Cancer Res 6:1557

    CAS  PubMed  Google Scholar 

  37. Pommier Y, Pourquier P, Fan Y, Strumberg D (1998) Mechanism of action of eukaryotic DNA topoisomerase I and drugs targeted to the enzyme. Biochim Biophys Acta 1400:83

    PubMed  Google Scholar 

  38. Prescott DM, Charles HC, Poulson JM, Page RL, Thrall DE, Vujaskovic Z, Dewhirst MW (2000) The relationship between intracellular and extracellular pH in spontaneous canine tumors. Clin Cancer Res 6:2501

    CAS  PubMed  Google Scholar 

  39. Raghunand N, He X, van Sluis R, Mahoney B, Baggett B, Taylor CW, Paine-Murrieta G, Roe D, Bhujwalla ZM, Gillies RJ (1999) Enhancement of chemotherapy by manipulation of tumour pH. Br J Cancer 80:1005

    Google Scholar 

  40. Robbins KT, Varki NM, Storniolo AM, Hoffman H, Hoffman RM (1991) Drug response of head and neck tumors in native-state histoculture. Arch Otolaryngol Head Neck Surg 117:83

    CAS  PubMed  Google Scholar 

  41. Robbins KT, Connors KM, Storniolo AM, Hanchett C, Hoffman RM (1994) Sponge-gel supported histoculture drug-response assay for head and neck cancer: correlations with clinical response to cisplatin. Arch Otolaryngol Head Neck Surg 120:288

    CAS  PubMed  Google Scholar 

  42. Rosen LS (1998) Irinotecan in lymphoma, leukemia, and breast, pancreatic, ovarian, and small-cell lung cancers. Oncology (Huntingt) 12:103

    Google Scholar 

  43. Ross DT, Scherf U, Eisen MB, Perou CM, Rees C, Spellman P, Iyer V, Jeffrey SS, Van de Rijn M, Waltham M, Pergamenschikov A, Lee JC, Lashkari D, Shalon D, Myers TG, Weinstein JN, Botstein D, Brown PO (2000) Systematic variation in gene expression patterns in human cancer cell lines. Nat Genet 24:227

    Article  CAS  PubMed  Google Scholar 

  44. Saramaki OR, Savinainen KJ, Nupponen NN, Bratt O, Visakorpi T (2001) Amplification of hypoxia-inducible factor 1alpha gene in prostate cancer. Cancer Genet Cytogenet 128:31

    CAS  PubMed  Google Scholar 

  45. Sawada S, Okajima S, Aiyama R, Nokata K, Furuta T, Yokokura T, Sugino E, Yamaguchi K, Miyasaka T (1991) Synthesis and antitumor activity of 20(S)-camptothecin derivatives: carbamate-linked, water-soluble derivatives of 7-ethyl-10-hydroxycamptothecin. Chem Pharm Bull (Tokyo) 39:1446

    Google Scholar 

  46. Singh B, Li R, Xu L, Poluri A, Patel S, Shaha AR, Pfister D, Sherman E, Goberdhan A, Hoffman RM, Shah J (2002) Prediction of survival in patients with head and neck cancer using the histoculture drug response assay. Head Neck 24:437

    Article  PubMed  Google Scholar 

  47. Theodorescu D, Connors KM, Groce A, Hoffman RM, Kerbel RS (1993) Lack of influence of c-Ha-ras expression on the drug sensitivity of human bladder cancer histocultured in three-dimensions. Anticancer Res 13:941

    CAS  PubMed  Google Scholar 

  48. Thistlethwaite AJ, Leeper DB, Moylan DJ 3rd, Nerlinger RE (1985) pH distribution in human tumors. Int J Radiat Oncol Biol Phys 11:1647

    Google Scholar 

  49. Vescio RA, Redfern CH, Nelson TJ, Ugoretz S, Stern PH, Hoffman RM (1987) In vivo-like drug responses of human tumors growing in three-dimensional gel-supported primary culture. Proc Natl Acad Sci U S A 84:5029

    PubMed  Google Scholar 

  50. Vescio RA, Connors KM, Youngkin T, Bordin GM, Robb JA, Umbreit JN, Hoffman RM (1990) Cancer biology for individualized therapy: correlation of growth fraction index in native-state histoculture with tumor grade and stage. Proc Natl Acad Sci U S A 87:691

    PubMed  Google Scholar 

  51. Vescio RA, Connors KM, Kubota T, Hoffman RM (1991) Correlation of histology and drug response of human tumors grown in native-state three-dimensional histoculture and in nude mice. Proc Natl Acad Sci U S A 88:5163

    CAS  PubMed  Google Scholar 

  52. Wachsberger PR, Landry J, Storck C, Davis K, O'Hara MD, Owen CS, Leeper DB, Coss RA (1997) Mammalian cells adapted to growth at pH 6.7 have elevated HSP27 levels and are resistant to cisplatin. Int J Hyperthermia 13:251

    CAS  PubMed  Google Scholar 

  53. Wall ME, Wani MC, Nicholas AW, Manikumar G, Tele C, Moore L, Truesdale A, Leitner P, Besterman JM (1993) Plant antitumor agents. 30. Synthesis and structure activity of novel camptothecin analogs. J Med Chem 36:2689

    CAS  PubMed  Google Scholar 

  54. Wallen JW, Cate RL, Kiefer DM, Riemen MW, Martinez D, Hoffman RM, Donahoe PK, Von Hoff DD, Pepinsky B, Oliff A (1989) Minimal antiproliferative effect of recombinant mullerian inhibiting substance on gynecological tumor cell lines and tumor explants. Cancer Res 49:2005

    CAS  PubMed  Google Scholar 

  55. Wani MC, Nicholas AW, Manikumar G, Wall ME (1987) Plant antitumor agents. 25. Total synthesis and antileukemic activity of ring A substituted camptothecin analogues. Structure-activity correlations. J Med Chem 30:1774

    CAS  PubMed  Google Scholar 

  56. Yoshimasu T, Oura S, Maebeya S, Tanino H, Bessho T, Arimoto J, Sakurai T, Matsuyama K, Naito Y, Furukawa T, Yano T, Suzuma T, Hirai I (2000) Histoculture drug response assay on non-small cell lung cancer (in Japanese). Gan To Kagaku Ryoho 27:717

    CAS  Google Scholar 

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Acknowledgements

This work is dedicated to the memory of Dr. Monroe E. Wall. The authors wish to acknowledge Alan Proia and Duke Surgical Pathology for invaluable assistance with procuring the primary human tissue samples and Stacey Snyder of Duke Radiation Oncology Department, Ray Liao and Dr. David Price of Duke Urology Department and Steve Keir of Duke Neurooncology Department for expert technical assistance and providing xenografts for histoculture experiments. This work was supported by NIH grant U01 CA68697-02.

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Authors and Affiliations

  1. Duke University Medical Center, Durham, NC 27710, USA

    James L. Flowers, Timothy A. Driscoll, Henry S. Friedman, Mark Dewhirst, O. Michael Colvin & David J. Adams

  2. AntiCancer, Inc., San Diego, CA 92111, USA

    Robert M. Hoffman

  3. Research Triangle Institute, Research Triangle Park, NC 27709, USA

    Monroe E. Wall, Mansukh C. Wani & Govindarajan Manikumar

  4. Department of Medicine, Duke University Medical Center, DUMC-3843, Durham, NC 27710, USA

    James L. Flowers

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  1. James L. Flowers
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  3. Timothy A. Driscoll
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  4. Monroe E. Wall
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Correspondence to David J. Adams.

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Flowers, J.L., Hoffman, R.M., Driscoll, T.A. et al. The activity of camptothecin analogues is enhanced in histocultures of human tumors and human tumor xenografts by modulation of extracellular pH. Cancer Chemother Pharmacol 52, 253–261 (2003). https://doi.org/10.1007/s00280-003-0635-7

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