BCc1, a Chelation-Based Nanomedicine, modulates splenic cytokine networks toward a profile associated with antitumor immunity in 4T1 breast tumor–bearing mice
Archives of Advances in Biosciences,
Vol. 17 No. 1 (2026),
28 April 2026
,
Page 1-13
https://doi.org/10.22037/aab.v16i1.52004
Abstract
Background and Aim: Breast cancer remains a major cause of cancer mortality in women, and tumor-driven cytokine dysregulation promotes immune escape. This study aimed to define the splenic immunomodulatory activity of BCc1 nanomedicine in 4T1 breast tumor-bearing BALB/c mice by measuring interleukin-12 (IL-12), transforming growth factor-β1 (TGF-β1), and tumor necrosis factor-α (TNF-α), and the (TGF-β1)⁄(IL-12) balance.
Methods: Seventy female mice with palpable tumors received a 24-day regimen of intraperitoneal or oral BCc1, cyclophosphamide, or phosphate-buffered saline (PBS). Splenic cytokines were quantified by ELISA, and the (TGF-β1)⁄(IL-12) ratio was calculated as an index of cytokine polarization.
Results: BCc1 reshaped splenic cytokine patterns in a dose- and route-dependent manner. High-dose intraperitoneal BCc1 produced the strongest effects, markedly reducing and , inducing controlled modulation of , and substantially lowering the ratio. These changes indicate reduced immunoregulatory dominance and a shift toward an antitumor-permissive cytokine milieu. For several cytokine endpoints, high-dose BCc1 showed effects comparable to or greater than cyclophosphamide. Oral BCc1 elicited milder dose-dependent responses, with stronger immunomodulation at the higher oral dose.
Conclusion: BCc1 therefore mediates route- and dose-dependent splenic cytokine remodeling, supporting further evaluation as a multifunctional immunomodulatory nanotherapeutic; cellular phenotyping and functional antitumor assays are required to determine whether these cytokine shifts translate into immune-mediated tumor control
Background and Aim: Breast cancer remains a major cause of cancer mortality in women, and tumor-driven cytokine dysregulation promotes immune escape. This study aimed to define the splenic immunomodulatory activity of BCc1 nanomedicine in 4T1 breast tumor-bearing BALB/c mice by measuring interleukin-12 (IL-12), transforming growth factor-β1 (TGF-β1), and tumor necrosis factor-α (TNF-α), and the (TGF-β1)⁄(IL-12) balance.
Methods: Seventy female mice with palpable tumors received a 24-day regimen of intraperitoneal or oral BCc1, cyclophosphamide, or phosphate-buffered saline (PBS). Splenic cytokines were quantified by ELISA, and the (TGF-β1)⁄(IL-12) ratio was calculated as an index of cytokine polarization.
Results: BCc1 reshaped splenic cytokine patterns in a dose- and route-dependent manner. High-dose intraperitoneal BCc1 produced the strongest effects, markedly reducing and , inducing controlled modulation of , and substantially lowering the ratio. These changes indicate reduced immunoregulatory dominance and a shift toward an antitumor-permissive cytokine milieu. For several cytokine endpoints, high-dose BCc1 showed effects comparable to or greater than cyclophosphamide. Oral BCc1 elicited milder dose-dependent responses, with stronger immunomodulation at the higher oral dose.
Conclusion: BCc1 therefore mediates route- and dose-dependent splenic cytokine remodeling, supporting further evaluation as a multifunctional immunomodulatory nanotherapeutic; cellular phenotyping and functional antitumor assays are required to determine whether these cytokine shifts translate into immune-mediated tumor control
- Chelation therapy
- Cytokine remodeling
- Immunomodulatory nanomedicine
- Tumor immune escape
- Splenic microenvironment
How to Cite
References
1. Yang F, He Q, Dai X, Zhang X, Song D. The potential role of nanomedicine in the treatment of breast cancer to overcome the obstacles of current therapies. Frontiers in Pharmacolgy. 2023 Feb;14:1143102. (DOI: 10.3389/fphar.2023.1143102) (PMID)
2. Lewis L, Thompson B, Stellmaker R, Koelmeyer L. Body composition and chemotherapy toxicities in breast cancer: a systematic review of the literature. Journal of Cancer Survivorship. 2025 Jun;19(3):914-29. (DOI: 10.1007/s11764-023-01512-z) (PMID)
3. Kitsios K, Sharifi S, Mahmoudi M. Nanomedicine technologies for diagnosis and treatment of breast cancer. ACS Pharmacology & Translational Science. 2023 Apr;6(5):671-82. (DOI: 10.1021/acsptsci.3c00044) (PMID)
4. Chaudhari R, Patel V, Kumar A. Cutting-edge approaches for targeted drug delivery in breast cancer: beyond conventional therapies. Nanoscale Advances. 2024 Apr;6(9):2270-86. (DOI: 10.1039/d4na00086b) (PMID)
5. Li M, Zhou S, Zhang Y, Li J, Zhang K. Advancements in tumor-targeted nanoparticles: Design strategies and multifunctional therapeutic approaches. Nanomaterials. 2025 Aug;15(16):1262. (DOI: 10.3390/nano15161262) (PMID)
6. Gao Z, Wan D, Luo M, Wei X. Application of nanomedicines in tumor immunotherapy. Journal of Molecular Cell Biology. 2025 Jun;16(12):mjae055. (DOI: 10.1093/jmcb/mjae055) (PMID)
7. Lin Y, Lin P, Chen X, Zhao X, Cui L. Harnessing nanoprodrugs to enhance cancer immunotherapy: overcoming barriers to precision treatment. Materials Today Bio. 2025 May;32:101933. (DOI: 10.1016/j.mtbio.2025.101933) (PMID)
8. Muradova Z, Carmès L, Brown N, Rossetti F, Guthier R, Yasmin-Karim S, et al. Targeted-theranostic nanoparticles induce anti-tumor immune response in lung cancer. Journal of Nanobiotechnology. 2025 Jul;23(1):466. (DOI: 10.1186/s12951-025-03542-4) (PMID)
9. Nazaran MH, inventor. Chelate compounds. United States patent US8288587B2. 2012.(LINK)
10. Kalanaky S, Hafizi M, Fakharzadeh S, Vasei M, Langroudi L, Janzamin E, et al. BCc1, the novel antineoplastic nanocomplex, showed potent anticancer effects in vitro and in vivo. Drug Design, Development and Therapy. 2015 Dec;10:59-70. (DOI: 10.2147/DDDT.S89694) (PMID)
11. Karimi P, Fakharzadeh S, Kalanaky S, Hafizi M, Hashemi M, Mahdavi M, et al. Immunologic Mechanisms of BCc1 Nanomedicine Synthesized by Nanochelating Technology in Breast Tumor-bearing Mice: Immunomodulation and Tumor Suppression. Anti-Cancer Agents in Medicinal Chemistry. 2024;24(19):1442-56. (DOI: 10.2174/0118715206302153240723053521) (PMID)
12. Hafizi M, Soleimani M, Noorian S, Kalanaky S, Fakharzadeh S, Tavakolpoor Saleh N, et al. Effects of BCc1 nanoparticle and its mixture with doxorubicin on survival of murine 4T1 tumor model. OncoTargets and Therapy. 2019 Jun;12:4691-4701. (DOI: 10.2147/OTT.S200446) (PMID)
13. Fakharzadeh S, Kalanaky S, Hafizi M, Goya MM, Masoumi Z, Namaki S, et al. The new nano-complex, Hep-c, improves the immunogenicity of the hepatitis B vaccine. Vaccine. 2013 May;31(22):2591-7. (DOI: 10.1016/j.vaccine.2013.03.030) (PMID)
14. Dong W, Li Y, Fei Q, Li S, He X, Chai Y, et al. Targeted spleen modulation: a novel strategy for next-generation disease immunotherapy. Theranostics. 2025 Mar;15(10):4416-45. (DOI: 10.7150/thno.111116) (PMID)
15. Pu T, Sun J, Ren G, Li H. Neuro-immune crosstalk in cancer: mechanisms and therapeutic implications. Signal Transduction and Targeted Therapy. 2025 Jun;10(1):176. (DOI: 10.1038/s41392-025-02241-8) (PMID)
16. Wang F, Lou J, Gao X, Zhang L, Sun F, Wang Z, et al. Spleen-targeted nanosystems for immunomodulation. Nano Today. 2023;52:101943. (DOI: 10.1016/j.nantod.2023.101943)
17. Gephart BD, Coulter DW, Solheim JC. Effects of the alkylating agent cyclophosphamide in potentiating anti-tumor immunity. International Journal of Molecular Sciences. 2025 Jul;26(13):6440. (DOI: 10.3390/ijms26136440) (PMID)
18. Turner PV, Brabb T, Pekow C, Vasbinder MA. Administration of substances to laboratory animals: routes of administration and factors to consider. Journal of the American Association for Laboratory Animal. 2011 Sep;50(5):600-13. (PMID)
19. Al Shoyaib A, Archie SR, Karamyan VT. Intraperitoneal route of drug administration: should it be used in experimental animal studies? Pharmaceutical Research. 2019 Dec;37(1):12. (DOI: 10.1007/s11095-019-2745-x) (PMID)
20. Landsiedel R, Hahn D, Ossig R, Ritz S, Sauer L, Buesen R, et al. Gut microbiome and plasma metabolome changes in rats after oral gavage of nanoparticles: sensitive indicators of possible adverse health effects. Particle and Fibre Toxicology. 2022 Mar;19(1):21. (DOI: 10.1186/s12989-022-00459-w) (PMID)
21. Diehl KH, Hull R, Morton D, Pfister R, Rabemampianina Y, Smith D, et al. A good practice guide to the administration of substances and removal of blood, including routes and volumes. Journal of Applied Toxicology. 2001 Jan-Feb;21(1):15-23. (DOI: 10.1002/jat.727) (PMID)
22. Yu L, Jin Y, Song M, Zhao Y, Zhang H. When natural compounds meet nanotechnology: nature-inspired nanomedicines for cancer immunotherapy. Pharmaceutics. 2022 Jul;14(8):1589. (DOI: 10.3390/pharmaceutics14081589) (PMID)
23. Kontoghiorghes GJ. Advances on chelation and chelator metal complexes in medicine. International Journal of Molecular Sciences. 2020 Apr;21(7):2499. (DOI: 10.3390/ijms21072499) (PMID)
24. Shah A, Dobrovolskaia MA. Immunological effects of iron oxide nanoparticles and iron-based complex drug formulations: Therapeutic benefits, toxicity, mechanistic insights, and translational considerations. Nanomedicine. 2018 Apr;14(3):977-990. (DOI: 10.1016/j.nano.2018.01.014) (PMID)
25. Palumbo GA, Galimberti S, Barcellini W, Cilloni D, Di Renzo N, Elli EM, et al. From biology to clinical practice: iron chelation therapy with deferasirox. Frontiers in Oncology. 2021 Oct;11:752192. (DOI: 10.3389/fonc.2021.752192) (PMID)
26. Hafizi M, Kalanaky S, Moaiery H, Khayamzadeh M, Noorian S, Kaveh V, et al. A randomized, double-blind, placebo-controlled investigation of BCc1 nanomedicine effect on survival and quality of life in metastatic and non-metastatic gastric cancer patients. Journal of Nanobiotechnology. 2019 Apr;17(1):52. (DOI: 10.1186/s12951-019-0484-0) (PMID)
27. Shigehiro T, Ueno M, Kijihira M, Takahashi R, Umemura C, Taha EA, et al. Immune state conversion of the mesenteric lymph node in a mouse breast cancer model. International Journal of Molecular Sciences. 2022 Sep;23(19):11035. (DOI: 10.3390/ijms231911035) (PMID)
28. Lewis SM, Williams A, Eisenbarth SC. Structure and function of the immune system in the spleen. Science Immunology. 2019 Mar;4(33):eaau6085. (DOI: 10.1126/sciimmunol.aau6085) (PMID)
29. Bronte V, Pittet MJ. The spleen in local and systemic regulation of immunity. Immunity. 2013 Nov;39(5):806-18. (DOI: 10.1016/j.immuni.2013.10.010) (PMID)
30. Lewis KL, Caton ML, Bogunovic M, Greter M, Grajkowska LT, Ng D, et al. Notch2 receptor signaling controls functional differentiation of dendritic cells in the spleen and intestine. Immunity. 2011 Nov;35(5):780-91. (DOI: 10.1016/j.immuni.2011.08.013) (PMID)
31. Teng MW, Bowman EP, McElwee JJ, Smyth MJ, Casanova J-L, Cooper AM, et al. IL-12 and IL-23 cytokines: from discovery to targeted therapies for immune-mediated inflammatory diseases. Nature Medicine. 2015 Jul;21(7):719-29. (DOI: 10.1038/nm.3895) (PMID)
32. Xu Y, Sun X, Tong Y. Interleukin-12 in multimodal tumor therapies for induction of anti-tumor immunity. Discover Oncology. 2024 May;15(1):170. (DOI: 10.1007/s12672-024-01011-2) (PMID)
33. Trinchieri G. Interleukin-12 and the regulation of innate resistance and adaptive immunity. Nature reviews immunology. 2003 Feb 1;3(2):133-46.(LINK)
34. Cirella A, Luri-Rey C, Di Trani CA, Teijeira A, Olivera I, Bolanos E, et al. Novel strategies exploiting interleukin-12 in cancer immunotherapy. Pharmacology & Therapeutics. 2022 Nov;239:108189. (DOI: 10.1016/j.pharmthera.2022.108189) (PMID)
35 Batlle E, Massagué J. Transforming growth factor-β signaling in immunity and cancer. Immunity. 2019 Apr 16;50(4):924-40.(LINK)
36. Bright JJ, Sriram S. TGF-β inhibits IL-12-induced activation of Jak-STAT pathway in T lymphocytes. Journal of Immunology. 1998 Aug;161(4):1772-7. (PMID)
37. Massagué J. TGFβ signalling in context. Nature reviews Molecular cell biology. 2012 Oct;13(10):616-30. (LINK)
38. Zhou M, Wang J, Pan J, Wang H, Huang L, Hou B, et al. Nanovesicles loaded with a TGF-β receptor 1 inhibitor overcome immune resistance to potentiate cancer immunotherapy. Nature communications. 2023 Jun;14(1):3593. (DOI: 10.1038/s41467-023-39035-x) (PMID)
39. Mariathasan S, Turley SJ, Nickles D, Castiglioni A, Yuen K, Wang Y, Kadel Iii EE, Koeppen H, Astarita JL, Cubas R, Jhunjhunwala S. TGFβ attenuates tumour response to PD-L1 blockade by contributing to exclusion of T cells. Nature. 2018 Feb 22;554(7693):544-8.(LINK)
40. Cruceriu D, Baldasici O, Balacescu O, Berindan-Neagoe I. The dual role of tumor necrosis factor-alpha (TNF-α) in breast cancer: molecular insights and therapeutic approaches. Cellular oncology (Dordrecht). 2020 Feb;43(1):1-18. (DOI: 10.1007/s13402-019-00489-1) (PMID)
41. Khan S, Aldawood Y, Shaikh AH, Zobairi A, Nabilah U, Alqahtani H, et al. Tumor Necrosis Factor-Alpha’s Role in the Pathophysiology of Colon Cancer. Diseases. 2025 Jun;13(6):185. (DOI: 10.3390/diseases13060185) (PMID)
42. Mussa A, Ismail NH, Hamid M, Al-Hatamleh MA, Bragoli A, Hajissa K, et al. Understanding the role of TNFR2 signaling in the tumor microenvironment of breast cancer. Journal of Experimental & Clinical Cancer Research. 2024 Nov;43(1):312. (DOI: 10.1186/s13046-024-03218-1) (PMID)
43. Afzali FM, Heshmati M, Salimi A, Kalanaky S, Fakharzadeh S, Hafizi M, et al. Nanochelate-based BCc1 delivery and its impact on key regulatory pathways in BALB/c breast cancer: An analysis of Beclin-1, ATG-4B, ATG-7, and mTOR expression. Biochemistry and Biophysics Reports. 2026 Jan;45:102418. (DOI: 10.1016/j.bbrep.2025.102418) (PMID)
- Abstract Viewed: 77 times
- PDF Downloaded: 12 times