Su Y, Zhang X, Bidlingmaier S, Behrens CR, Lee NK, Liu B (2020) ALPPL2 is a highly specific and targetable tumor cell surface antigen. Cancer Res 80(20):4552-4564. doi: 10.1158/0008-5472.CAN-20-1418 PMID: 32868383
Objective: To evaluate therapeutic potential of ALPPL2 targeting.
Summary: Exquisite tissue specificity and broad tumor type coverage suggest that ALPPL2 could be an excellent cell surface target for therapeutic development against mesothelioma.
Usage: Biotinylated M25 IgG1 and Streptavidin-ZAP were mixed at a molar ratio of 1:1.
Rizvanovic H, Pinheiro AD, Kim K, Thomas J (2019) Chlorotoxin conjugated with saporin reduces viability of ML-1 thyroid cancer cells in vitro. bioRxiv 885483. doi: 10.1101/2019.12.20.885483
Objective: To determine whether Chlorotoxin-conjugated Saporin (CTX-SAP) would inhibit the growth of aggressive thyroid cancer cell lines expressing MMP-2.
Summary: This in vitro study demonstrated the efficacy of a CTX-SAP conjugate in reducing the viability of ML-1 thyroid cancer cells in a dose dependent manner.
Usage: There was a statistically significant reduction in cell viability with increasing concentrations of the CTX-SAP conjugate (biotinylated Chlorotoxin mixed with Streptavidin-ZAP). The cell viability of ML-1 cells was decreased by 49.77% with the treatment of 600 nM of CTX-SAP (F=44.24). Unconjugated Chlorotoxin or Saporin had no significant effect on cell viability a using similar assay.
Zurlo G, Liu X, Takada M, Fan C, Simon JM, Ptacek TS, Rodriguez J, von Kriegsheim A, Liu J, Locasale JW, Robinson A, Zhang J, Holler JM, Kim B, Zikánová M, Bierau J, Xie L, Chen X, Li M, Perou CM, Zhang Q (2019) Prolyl hydroxylase substrate adenylosuccinate lyase is an oncogenic driver in triple negative breast cancer. Nat Commun 10(1):5177. doi: 10.1038/s41467-019-13168-4 PMID: 31729379
Objective: To investigate the role of ADSL hydroxylation in controlling cMYC and triple negative breast cancer (TNBC) tumorigenesis.
Summary: ADSL regulates cMYC protein level through adenosine levels and leads to downstream metabolic changes and breast cancer cell proliferation. ADSL is essential for TNBC cell growth and invasiveness and contributes to TNBC breast tumorigenesis and metastasis in vivo.
Wu SJ, Coarfa C, Abbas H, Checker R, Dhar S, Rajapakshe K, Lee MG, Flores ER (2019) ∆Np63 executes an oncogenic program through the regulation of a common landscape of super-enhancer associated genes in non-small cell lung cancer. Cell Reports 3420369. doi: 10.2139/ssrn.3420369
Objective: To investigate how ΔNp63 serves to maintain lung adenocarcinoma (ADC) and squamous cell carcinoma (SCC) through the regulation and maintenance of lung stem cells that are necessary for lung adenocarcinoma and squamous cell initiation and progression.
Summary: ∆Np63 maintains lung ADC and SCC by keeping lung stem cells in quiescence. ChIP-seq analysis of lung basal cells and alveolar type 2 (AT2) cells lacking ∆Np63 revealed a robust loss of activating histone marks at super enhancers of cell identity genes defining a unifying oncogenic role for ∆Np63 in non-small cell lung cancer.
Thorén MM, Chmielarska Masoumi K, Krona C, Huang X, Kundu S, Schmidt L, Forsberg-Nilsson K, Floyd Keep M, Englund E, Nelander S, Holmqvist B, Lundgren-Åkerlund E (2019) Integrin α10, a novel therapeutic target in glioblastoma, regulates cell migration, proliferation, and survival. Cancers (Basel) 11(4):587. doi: 10.3390/cancers11040587 PMID: 31027305
Objective: To investigate the potential of integrin alpha10beta1 as a therapeutic target in glioblastomas (GBMs).
Summary: Integrin alpha10beta1 has a crucial role in the migration, proliferation, and survival of GBM cells and that an integrin alpha10beta1 antibody–drug conjugate induced cell death of GBM cells both in vitro and in vivo.
Usage: Infusions of anti- 10-SAP or Anti-ctrl-SAP were made icv (1 mcg/2 L per infusion).
Shramm PA, Ancheta L (2019) Streptavidin-pHast: A readily conjugatable, pH-sensitive dye to screen for internalization. Cancer Res 79(13 Suppl):2177. Proceedings of the American Association for Cancer Research Annual Meeting, Atlanta GA. PMID: 909090
Summary: Quick and efficient screening of targeting agents that internalize effectively is vital for determining their suitability as potential therapeutics. Some of the most recent successes in the treatment of cancers have been from antibodies to cell surface proteins that are responsible for tumor cell proliferation. Examples are Cetuximab (target: EGFR) approved for colorectal cancer, and Trastuzumab (target: HER2) for breast cancer. These antibodies have more than one effect on the cancer cell, but one of the most important is that, upon binding to the cell surface antigen, the complex is internalized. As such, the down-regulated cell surface protein no longer plays a role in cancer cell division. Here we describe a method for determining internalization of cell surface molecules by targeting agents using a pH-dependent fluorescent reporter cross-linked to streptavidin. Streptavidin is a tetrameric protein (molecular weight 53 kDa in its recombinant form), with each subunit able to bind a single biotin molecule. The bond between streptavidin and biotin is rapid and essentially non-reversible, unaffected by most extremes of pH, organic solvents, and denaturing reagents. It is the strongest known noncovalent biological interaction (Ka = 1015 M-1) between protein and ligand. A variety of molecules, including lectins, proteins, and antibodies, can be biotinylated and reacted with streptavidin-labeled probes or other detection reagents for use in biological assays. The fluorescence from this reporter increases intensity as the pH of its surroundings becomes more acidic, as demonstrated when exposed to the environment inside a cell (thereby providing evidence of internalization). Here we describe methods that can be used to explore candidates as cancer therapeutics in a quick, reliable and reproducible manner.
Im E-J, Lee C-H, Moon P-G, Rangaswamy GG, Lee B, Lee JM, Lee J-C, Jee J-G, Bae J-S, Kwon T-K, Kang K-W, Jeong M-S, Lee J-E, Jung H-S, Ro H-J, Jun S, Kang W, Seo S-Y, Cho Y-E, Song B-J, Baek M-C (2019) Sulfisoxazole inhibits the secretion of small extracellular vesicles by targeting the endothelin receptor A. Nature Commun 10(1):1387. doi: 10.1038/s41467-019-09387-4 PMID: 30918259
Objective: To identify a direct protein target of SFX in cancer cells.
Summary: Using data from the BindingDB, SEA identified four proteins as target candidates, which are: endothelin receptor type A, kynurenine 3-monooxygenase, carbonic anhydrase 13, and angiotensin II type 1a receptor.
Zangardi ML, Spring LM, Nagayama A, Bardia A (2019) Sacituzumab for the treatment of triple-negative breast cancer: the poster child of future therapy?. Expert Opin Investig Drugs 28(2):107-112. doi: 10.1080/13543784.2019.1555239 PMID: 30507322
Summary: This publication explores the potential of sacituzumab as a treatment for triple-negative breast cancer, highlighting its promising role as a leading candidate for future therapeutic approaches. The study suggests that sacituzumab may represent a breakthrough in the management of this aggressive breast cancer subtype, raising hopes for improved outcomes in patients.
Berhani O, Glasner A, Kahlon S, Duev-Cohen A, Yamin R, Horwitz E, Enk J, Moshel O, Varvak A, Porgador A, Jonjic S, Mandelboim O. (2019) Human anti-NKp46 antibody for studies of NKp46-dependent NK cell function and its applications for type 1 diabetes and cancer research. Eur J Immunol 49(2):228-241. doi: 10.1002/eji.201847611 PMID: 30536875
Objective: To investigate human NKp46 activity and its critical role in Natural Killer (NK) cell biology.
Summary: A unique anti-human NKp46 monoclocal antibody was developed and conjugated to Saporin. Targeted toxin inhibits growth of NKp46-positive cells; thus, exemplifying the potential as an immunotherapeutic drug to treat NKp46-dependent diseases, such as, type I diabetes and NK and T cell related malignancies.
Usage: Conjugation of the antibodies to Saporin, treatment of cells, and cell viability assay Biotin-Z Kit instructions.
Kinneer K, Meekin J, Tiberghien AC, Tai YT, Phipps S, Kiefer CM, Rebelatto MC, Dimasi N, Moriarty A, Papadopoulos KP, Sridhar S, Gregson SJ, Wick MJ, Masterson L, Anderson KC, Herbst R, Howard PW, Tice DA (2018) SLC46A3 as a potential predictive biomarker for antibody-drug conjugates bearing noncleavable linked maytansinoid and pyrrolobenzodiazepine warheads. Clin Cancer Res 24(24):6570-6582. doi: 10.1158/1078-0432.CCR-18-1300 PMID: 30131388
Objective: To develop biomarkers to uncover the underlying mechanism of resistance by certain cell lines for ADCs.
Summary: Loss of SLC46A3 expression was found to be a mechanism of innate and acquired resistance to ADCs bearing DM1 and SG3376.
Usage: For Lysosomal trafficking, ADCs were labeled with Fab-pHast human (Cat. #PH-01). Cells were incubated with 3 mg/mL of labeled ADCs at 37°C for desired time points and fluorescence quantified by flow cytometry.
