carbocyanines and Burkitt-Lymphoma

This review covers the application of heptamethine cyanine dye (HMCD) mediated drug delivery. A relatively small number of HMCDs possess tumor targeting abilities, and this has spurred interest from research groups to explore them as drug delivery systems. Their tumor selectivity is primarily attributed to their uptake by certain isoforms of organic anion transporting polypeptides (OATPs) which are overexpressed in cancer tissues, although there are other possible mechanisms for the observed selectivity still under investigation. This specificity is confirmed using various cancer cell lines and is accompanied by moderate cytotoxicity. Their retention in tumor tissue is facilitated by the formation of albumin adducts as revealed by published mechanistic studies. HMCDs are also organelle selective dyes with specificity toward mitochondria and lysosomes, and with absorption and emission in the near-infrared region. This makes them valuable tools for biomedical imaging, especially in the field of fluorescence-guided tumor surgery. Furthermore, conjugating antitumor agents to HMCDs is providing novel drugs that await clinical testing. HMCD development as theranostic agents with dual tumor targeting and treatment capability signals a new approach to overcome drug resistance (mediated through evasion of efflux pumps) and systemic toxicity, the two parameters which have long plagued drug discovery.

Topics: Antineoplastic Agents; Brain Neoplasms; Breast Neoplasms; Burkitt Lymphoma; Carbocyanines; Coloring Agents; Drug Delivery Systems; Drug Discovery; Drug Resistance, Neoplasm; Female; Humans; Kidney Neoplasms; Male; Precision Medicine; Prostatic Neoplasms

Burkitt lymphoma is a fast-growing mature B cell malignancy, whose genetic hallmark is translocation and activation of the c-myc gene. Prompt multiagent immunochemotherapy regimens can have favorable outcomes, but prognosis is poor in refractory or relapsed disease. We previously identified a novel family of near-infrared heptamethine carbocyanine fluorescent dyes (HMCD or DZ) with tumor-homing properties via organic anion-transporting peptides. These membrane carriers have uptake in tumor cells but not normal cells in cell culture, mouse and dog tumor models, patient-derived xenografts, and perfused kidney cancers in human patients.. Here we report the cytotoxic effects of a synthesized conjugate of DZ with cisplatin (CIS) on B cell lymphoma CA46, Daudi, Namalwa, Raji, and Ramos cell lines in cell culture and in xenograft tumor formation. Impaired mitochondrial membrane permeability was examined as the mechanism of DZ-CIS-induced lymphoma cell death.. The new conjugate, DZ-CIS, is cytotoxic against Burkitt lymphoma cell lines and tumor models. DZ-CIS retains tumor-homing properties to mitochondrial and lysosomal compartments, does not accumulate in normal cells and tissues, and has no nephrotoxicity in mice. DZ-CIS accumulated in Burkitt lymphoma cells and tumors induces apoptosis and retards tumor cell growth in culture and xenograft tumor growth in mice.. DZ-CIS downregulated c-myc and overcame CIS resistance in myc-driven TP53-mutated aggressive B cell Burkitt lymphoma. We propose that DZ-CIS could be used to treat relapsed/refractory aggressive Burkitt lymphomas.

Topics: Animals; Antineoplastic Agents; Apoptosis; Burkitt Lymphoma; Carbocyanines; Cell Proliferation; Cisplatin; Drug Compounding; Humans; Male; Mice; Mice, Inbred NOD; Mice, SCID; Tumor Cells, Cultured; Xenograft Model Antitumor Assays

Detection of grafted human cells in mice using fluorescence is a rapid and simple technique whose use is continually expanding. Robust engraftment of human hematological malignancy (HHM) lines and patient cells into the naturally immunodeficient turkey embryo has recently been demonstrated by polymerase chain reaction (PCR), fluorescence activated cell sorting (FACS) and histology. We demonstrate here that fluorescence imaging is a rapid and simple technique for detecting engraftment and homing of cells derived from HHM in turkey embryos. Raji lymphoma cells expressing a far-red fluorescent protein were injected intravascularly into turkey embryos and fluorescence was detected 8 days later in their limbs and skulls. Much stronger signals were obtained after removal of the bones from the limbs. Unlabeled Raji cells did not give a fluorescent signal. Treatment with doxorubicin dramatically reduced the fluorescent signal. Intravenously injected HL-60 leukemia cells labeled with infrared-fluorescing dye were detected in the bone marrow after 16 h. Homing was active, although some non-specific fluorescence was present. Use of fluorescence imaging of HHM in turkey embryos is therefore feasible and reduces the time, effort and expense for detecting engraftment. This technique has potential to become a high-throughput xenograft system for hematological chemotherapy development and testing, and for study of hematological cell homing.

Topics: Animals; Bone Marrow; Bone Marrow Cells; Burkitt Lymphoma; Carbocyanines; Cell Line, Tumor; Cell Lineage; Cell Movement; Cell Separation; Doxorubicin; Embryo, Nonmammalian; Fluorescent Dyes; Graft Survival; Green Fluorescent Proteins; HL-60 Cells; Humans; Luminescent Proteins; Microscopy, Fluorescence; Neoplasm Transplantation; Turkeys; Xenograft Model Antitumor Assays

Overexpression of human polμ in a Burkitt's lymphoma-derived B cell line (RAMOS), in which somatic hypermutation (SHM) is constitutive, induced an increase in somatic mutations in the parental cell line (Nucleic Acids Res 32:5861-5873, 2004). To further study Polμ implications in SHM, a dominant-negative (DN) mutant of Polμ (Polμ-DN) was generated which showed moderated overexpression of the Polμ-DN protein. The subcellular prefractionation was used to improve the detection of low-abundance proteins contained in membrane/organelles and nuclei, which are efficiently separated from high-abundance proteins commonly found in the cytosol that might otherwise hamper analysis. Two-dimensional (2D) difference gel electrophoresis (DIGE) is a technique for comparative proteomics, which improves the reproducibility and reliability of differential protein expression analysis between samples. The standard sample included in every gel (Cy2) comprises equal amounts of each sample to be compared, and thus improves the accuracy of protein quantification between samples from different gels, allowing accurate detection of small differences in protein levels. The combination of this techniques allowed the detection in Fraction F2 (membrane/organelles) of 2,111 spots, 55 of them with significant variation (19 increased and 36 decreased in a ratio >2.0 or <-2.0), and in Fraction F3 (nuclear) of 2,416 spots, 80 of them with significant variation (51 increased and 29 decreased in a ratio >1.5 or <-1.5).

Topics: Analytic Sample Preparation Methods; B-Lymphocytes; Burkitt Lymphoma; Carbocyanines; Cell Line, Tumor; Humans; Image Processing, Computer-Assisted; Intracellular Space; Isoelectric Focusing; Proteome; Solubility; Staining and Labeling; Two-Dimensional Difference Gel Electrophoresis