Induction of cyclophosphamide-resistance by aldehyde-dehydrogenase gene transfer

Department of Pathology, Columbia University, New York, NY 10032, USA.
Blood (Impact Factor: 10.45). 03/1996; 87(3):1097-103.
Source: PubMed


The identification of genes inducing resistance to anticancer chemotherapeutic agents and their introduction into hematopoietic cells represents a promising approach to overcome bone marrow toxicity, the limiting factor for most high-dose chemotherapy regimens. Because resistance to cyclophosphamide has been correlated with increased levels of expression of the aldehyde-dehydrogenase (ALDH1) gene in tumor cell lines in vitro, we tested whether ALDH1 overexpression could directly induce cyclophosphamide resistance. We have cloned a full-length human ALDH1 cDNA and used retroviral vectors to transduce it into human (U937) and murine (L1210) hematopoietic cell lines that were then tested for resistance to maphosphamide, an active analogue of cyclophosphamide. Overexpression of the ALDH1 gene resulted in a significant increases in cyclophosphamide resistance in transduced L1210 and U937 cells (50% inhibition concentration [IC50], approximately 13 mumol/L). The resistant phenotype was specifically caused by ALDH1 overexpression as shown by its reversion by disulfiram, a specific ALDH1 inhibitor. ALDH1 transduction into peripheral blood human hematopoietic progenitor cells also led to significant increases (4- to 10-fold; IC50, approximately 3 to 4 mumol/L) in cyclophosphamide resistance in an in vitro colony-forming assay. These findings indicate that ALDH1 overexpression is sufficient to induce cyclophosphamide resistance in vitro and provide a basis for testing the efficacy of ALDH1 gene transduction to protect bone marrow cells from high-dose cyclophosphamide in vivo.

32 Reads
  • Source
    • "Increased activity of aldehyde dehydrogenase (ALDH), a detoxifying enzyme responsible for the oxidation of intracellular aldehydes [24], [25], has been detected in some stem/progenitor cells. For example, high ALDH activity has been found in murine and human hematopoietic and neural stem and progenitor cells [26]–[29]. "
    [Show abstract] [Hide abstract]
    ABSTRACT: High levels of aldehyde dehydrogenase (ALDH) activity have been detected in various progenitor and stem cells. Thus, Aldefluor fluorescence, which represents precisely the ALDH activity, has been widely used for the identification, evaluation, and isolation of stem and progenitor cells. Recently, ALDH activity was detected in embryonic and adult mouse pancreas, specifically in adult centroacinar and terminal duct cells supposed to harbor endocrine and exocrine progenitor cells in the adult pancreas. Nevertheless, ALDH activity and aldeflour fluorescence have not been examined in beta cells. Here, we report a dynamic increase in the number of aldeflour+ beta cells during pregnancy. Interestingly, nearly all these aldeflour+ beta cells are positive for Ki-67, suggesting that they are in an active cell cycle (G1, S and M phases). To determine precisely at which phase beta cells activate ALDH activity and thus become aldeflour+, we co-stained insulin with additional proliferation markers, phosphohistone3 (PHH3, a marker for M-phase proliferating cells) and Bromodeoxyuridine (BrdU, a marker for S-phase proliferating cells). Our data show little aldeflour+ beta cells that were positive for either PHH3, or BrdU, suggesting that beta cells activate ALDH and become Aldefluor+ when they enter G1-phase of active cell cycle, but may downregulate ALDH when they leave G1-phase and enter S phase. Our data thus reveal a potential change in ALDH activity of proliferating beta cells during pregnancy, which provides a novel method for isolation and analysis of proliferating beta cells. Moreover, our data also suggest that caution needs to be taken on interpretation of Aldefluor lineage-tracing data in pancreas.
    PLoS ONE 05/2014; 9(5):e96204. DOI:10.1371/journal.pone.0096204 · 3.23 Impact Factor
  • Source
    • "ALDHs contribute to drug resistance through detoxification of many cytotoxic agents provided that aldehydes are generated by several metabolic processes (reviewed by Marchitti et al.[156]). Increased ALDH activity in hematopoietic stem cells, for example, contributes to metabolize and detoxify cyclophosphamide [157]. The ALDH family of enzymes comprises 19 isoforms that can be found in different cell compartments: nucleus, cytoplasm or mitochondria. "
    [Show abstract] [Hide abstract]
    ABSTRACT: Some cancers like melanoma and pancreatic and ovarian cancers, for example, commonly display resistance to chemotherapy, and this is the major obstacle to a better prognosis of patients. Frequently, literature presents studies in monolayer cell cultures, 3D cell cultures or in vivo studies, but rarely the same work compares results of drug resistance in different models. Several of these works are presented in this review and show that usually cells in 3D culture are more resistant to drugs than monolayer cultured cells due to different mechanisms. Searching for new strategies to sensitize different tumors to chemotherapy, many methods have been studied to understand the mechanisms whereby cancer cells acquire drug resistance. These methods have been strongly advanced along the years and therapies using different drugs have been increasingly proposed to induce cell death in resistant cells of different cancers. Recently, cancer stem cells (CSCs) have been extensively studied because they would be the only cells capable of sustaining tumorigenesis. It is believed that the resistance of CSCs to currently used chemotherapeutics is a major contributing factor in cancer recurrence and later metastasis development. This review aims to appraise the experimental progress in the study of acquired drug resistance of cancer cells in different models as well as to understand the role of CSCs as the major contributing factor in cancer recurrence and metastasis development, describing how CSCs can be identified and isolated.
    Journal of Experimental & Clinical Cancer Research 04/2014; 33(1):37. DOI:10.1186/1756-9966-33-37 · 4.43 Impact Factor
  • Source
    • "Aldehyde dehydrogenase-1 (ALDH1) is a cytosolic enzyme, responsible for the metabolism of intracellular aldehydes [16]. Numerous preclinical studies have shown that expression of ALDH1 in tumor cells has been associated with stem-like characteristics, including innate chemoresistance and clonal capacity [17-20]. Moreover, expression of ALDH1 in surgically resected breast cancer is strongly associated with metastasis and poor survival [18,21]. "
    [Show abstract] [Hide abstract]
    ABSTRACT: Although neoadjuvant chemotherapy (NAC) for locally advanced breast cancer can improve operability and local disease control, there is a lack of reliable biomarkers that predict response to chemotherapy or long-term survival. Since expression of ALDEHYDE DEHYDROGENASE-1 (ALDH1) is associated with the stem-like properties of self-renewal and innate chemoresistance in breast cancer, we asked whether expression in serial tumour samples treated with NAC could identify women more likely to benefit from this therapy. Women with locally advanced breast cancer were randomly assigned to receive four cycles of anthracycline-based chemotherapy, followed by four cycles of taxane therapy (Arm A), or the same regimen in reverse order (Arm B). Tumour specimens were collected at baseline, after four cycles, and then at surgical resection. ALDH1 expression was determined by immunohistochemistry and correlated with tumour response using Fisher's exact test while Kaplan-Meier method was used to calculate survival. A hundred and nineteen women were enrolled into the study. Fifty seven (48%) were randomized to Arm A and 62 (52%) to Arm B. Most of the women (90%) had ductal carcinoma and 10% had lobular carcinoma. Of these, 26 (22%) achieved a pathological complete response (pCR) after NAC. There was no correlation between baseline ALDH1 expression and tumour grade, stage, hormone receptor, HER2 status and Ki67 index. ALDH1 negativity at baseline was significantly associated with cPR (p = 0.004). The presence of ALDH1(+) cells in the residual tumour cells in non-responding women was strongly predictive of worse overall survival (p = 0.024). Moreover, serial analysis of specimens from non-responders showed a marked increase in tumour-specific ALDH1 expression (p = 0.028). Overall, there was no survival difference according to the chemotherapy sequence. However, poorly responding tumours from women receiving docetaxel chemotherapy showed an unexpected significant increase in ALDH1 expression. ALDH1 expression is a useful predictor of chemoresistance. The upregulation of ALDH1 after NAC predicts for poor survival in locally advanced breast cancer. Although the chemotherapy sequence had no effect on overall prognosis, our results suggest that anthracycline-based chemotherapy may be more effective at targeting ALDH1(+) breast cancer cells.Trial registration: ACTRN12605000588695.
    Breast cancer research: BCR 04/2014; 16(2):R44. DOI:10.1186/bcr3648 · 5.49 Impact Factor
Show more


32 Reads