Abstract
The concept of resilience, defined as the ability to recover from stress, is a potential platform to predict healthy aging. However, specific stress tests for resilience have not yet been fully established in humans so investigations in animal models are of interest. The chemotherapeutic drug cyclophosphamide (Cyp) was selected as a chemical stressor to investigate resilience response in C57Bl/6 male mice at 4, 15, and 28 months of age. Following a single intraperitoneal injection of Cyp (100 mg/kg), tail blood was collected for counting white blood cells (WBC) every other day for 25 days, and physiological performance tests performed. Cyp induced a consistent pattern in neutrophil count in all three age groups, with a nadir at day 5 and a rebound at day 7 with different rates in each group. The neutrophil to lymphocyte ratio (NLR) showed an age-dependent rebound response 7 days after Cyp injection, with a similar pattern of decline back toward baseline. Mice in the 15-month age group with high pre-injection Cyp NLR had significantly higher total WBC counts after Cyp injection compared with mice with low pre-injection Cyp NLR, indicating a correlation between NLR and Cyp-altered WBC counts. In addition, mice with high pre-injection Cyp NLR showed significant learning impairment compared with mice with low pre-injection Cyp NLR, suggesting low NRL intensity can predict resilience to age-related cognitive decline. These observations provide the rationale to translate findings from the mouse to humans in developing in vitro Cyp stress tests.
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References
Allen RS, Dorman HR, Henkin H, Carden KD, Potts D. Definition of resilience. In: Resnick B, Gwyther LP, Roberto KA, editors. Resilience in aging: concepts, research, and outcomes. Cham: Springer International Publishing; 2018. p. 1–15.
Baguley BC, Beland FA, Berger MR, et al. A review of human carcinogens. Iarc, France: Pharmaceuticals; 2008.
Bjorner M, Zhu L. A minimally invasive, low-stress method for serial blood collection in aging mice. Pathobiol Aging Age-related Dis. 2019;9:1647400. https://doi.org/10.1080/20010001.2019.1647400.
Blagosklonny MV. Aging and immortality: quasi-programmed senescence and its pharmacologic inhibition. Cell Cycle. 2006;5:2087–102. https://doi.org/10.4161/cc.5.18.3288.
Christie LA, Acharya MM, Parihar VK, Nguyen A, Martirosian V, Limoli CL. Impaired cognitive function and hippocampal neurogenesis following cancer chemotherapy. Clin Cancer Res. 2012;18:1954–65. https://doi.org/10.1158/1078-0432.CCR-11-2000.
Darvas M, Mukherjee K, Lee A, Ladiges W. A novel one-day learning procedure for mice. Curr Protoc Mouse Biol. 2020;10:1–6. https://doi.org/10.1002/cpmo.68.
Demaria M, O’Leary MN, Chang J, et al. Cellular senescence promotes adverse effects of chemotherapy and cancer relapse. Cancer Discov. 2017;7:165–76. https://doi.org/10.1158/2159-8290.CD-16-0241.
Doloff JC, Waxman DJ. Transcriptional profiling provides insights into metronomic cyclophosphamide-activated, innate immune-dependent regression of brain tumor xenografts. BMC Cancer. 2015;15:375. https://doi.org/10.1186/s12885-015-1358-y.
Emmenegger U, Man S, Shaked Y, Francia G, Wong JW, Hicklin DJ, et al. A comparative analysis of low-dose metronomic cyclophosphamide reveals absent or low-grade toxicity on tissues highly sensitive to the toxic effects of maximum tolerated dose regimens. Cancer Res. 2004;64:3994–4000. https://doi.org/10.1158/0008-5472.CAN-04-0580.
Janelsins MC, Heckler CE, Thompson BD, Gross RA, Opanashuk LA, Cory-Slechta DA. A clinically relevant dose of cyclophosphamide chemotherapy impairs memory performance on the delayed spatial alternation task that is sustained over time as mice age. Neurotoxicology. 2016;56:287–93. https://doi.org/10.1016/j.neuro.2016.06.013.
Janssen-Heijnen MLG, Van Spronsen DJ, Lemmens VEPP, et al. A population-based study of severity of comorbidity among patients with non-Hodgkin’s lymphoma: prognostic impact independent of international prognostic index. Br J Haematol. 2005;129:597–606. https://doi.org/10.1111/j.1365-2141.2005.05508.x.
Kanakry CG, Fuchs EJ, Luznik L. Modern approaches to HLA-haploidentical blood or marrow transplantation. Nat Rev Clin Oncol. 2016;13:10–24. https://doi.org/10.1038/nrclinonc.2015.128.
Kato A, Sugiura T, Yamamoto T, Misaki T, Tsuji T, Sakao Y, et al. Water intoxication induced by low-dose oral cyclophosphamide in a patient with anti-neutrophil cytoplasmic antibody-related glomerulonephritis. NDT Plus. 2008;1:286–8. https://doi.org/10.1093/ndtplus/sfn076.
Kato K, Takeuchi A, Akashi K, Eto M. Cyclophosphamide-induced tolerance in allogeneic transplantation: from basic studies to clinical application. Front Immunol. 2020;10:1–8. https://doi.org/10.3389/fimmu.2019.03138.
Kirkland JL, Stout MB, Sierra F. Resilience in aging mice. Journals Gerontol - Ser A Biol Sci Med Sci. 2016;71:1407–14. https://doi.org/10.1093/gerona/glw086.
Li J, Chen Q, Luo X, Hong J, Pan K, Lin X, et al. Neutrophil-to-lymphocyte ratio positively correlates to age in healthy population. J Clin Lab Anal. 2015;29:437–43. https://doi.org/10.1002/jcla.21791.
Liu F, Poursine-Laurent J, Link DC. The granulocyte colony-stimulating factor receptor is required for the mobilization of murine hematopoietic progenitors into peripheral blood by cyclophosphamide or interleukin-8 but not Flt-3 ligand. Blood. 1997;90:2522–8. https://doi.org/10.1182/blood.v90.7.2522.
López-Otín C, Blasco MA, Partridge L, Serrano M, Kroemer G. The hallmarks of aging. Cell. 2013;153:1194–217. https://doi.org/10.1016/j.cell.2013.05.039.
MacLeod JE, DeLeo JA, Hickey WF, et al. Cancer chemotherapy impairs contextual but not cue-specific fear memory. Behav Brain Res. 2007;181:168–72. https://doi.org/10.1016/j.bbr.2007.04.003.
Miller KD, Nogueira L, Mariotto AB, Rowland JH, Yabroff KR, Alfano CM, et al. Cancer treatment and survivorship statistics, 2019. CA Cancer J Clin. 2019;69:363–85. https://doi.org/10.3322/caac.21565.
O’Connell KE, Mikkola AM, Stepanek AM, Vernet A, Hall CD, Sun CC, et al. Practical murine hematopathology: a comparative review and implications for research. Comp Med. 2015;65(2):96–113.
Panis C, Herrera ACSA, Victorino VJ, Campos FC, Freitas LF, de Rossi T, et al. Oxidative stress and hematological profiles of advanced breast cancer patients subjected to paclitaxel or doxorubicin chemotherapy. Breast Cancer Res Treat. 2012;133:89–97. https://doi.org/10.1007/s10549-011-1693-x.
Schorr A, Carter C, Ladiges W. The potential use of physical resilience to predict healthy aging. Pathobiol Aging Age-related Dis. 2017;8:1403844. https://doi.org/10.1080/20010001.2017.1403844.
Šefc L, Pšenák O, Sýkora V, Šulc K, Nečas E. Response of hematopoiesis to cyclophosphamide follows highly specific patterns in bone marrow and spleen. J Hematotherapy Stem Cell Res. 2003;12:47–61. https://doi.org/10.1089/152581603321210136.
Valiathan R, Ashman M, Asthana D. Effects of ageing on the immune system: infants to elderly. Scand J Immunol. 2016;83:255–66. https://doi.org/10.1111/sji.12413.
Watters JW, Kloss EF, Link DC, Graubert TA, McLeod HL. A mouse-based strategy for cyclophosphamide pharmacogenomic discovery. J Appl Physiol. 2003;95:1352–60. https://doi.org/10.1152/japplphysiol.00214.2003.
Wei Z, Wang D, Luo D, et al. Intervention of low-dose cyclophosphamide on neutrophil in rat bone marrow at early postburn stage. J Clin Rehabil Tissue Eng Res. 2007;11:8541–4.
Wilson BE, Jacob S, Yap ML, Ferlay J, Bray F, Barton MB. Estimates of global chemotherapy demands and corresponding physician workforce requirements for 2018 and 2040: a population-based study. Lancet Oncol. 2019;20:769–80. https://doi.org/10.1016/S1470-2045(19)30163-9.
Winkler IG, Pettit AR, Raggatt LJ, Jacobsen RN, Forristal CE, Barbier V, et al. Hematopoietic stem cell mobilizing agents G-CSF, cyclophosphamide or AMD3100 have distinct mechanisms of action on bone marrow HSC niches and bone formation. Leukemia. 2012;26:1594–601. https://doi.org/10.1038/leu.2012.17.
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This work was supported by NIA grant R01 AG057381 (Ladiges, PI).
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Zhu, L., Dou, Y., Bjorner, M. et al. Development of a cyclophosphamide stress test to predict resilience to aging in mice. GeroScience 42, 1675–1683 (2020). https://doi.org/10.1007/s11357-020-00222-z
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DOI: https://doi.org/10.1007/s11357-020-00222-z