Abstract
A deeper pathophysiologic understanding of available mouse models of sickle cell disease
(SCD), such as the Townes model, will help improve preclinical studies. We evaluated
groups of Townes mice expressing either normal adult human hemoglobin (HbA), sickle
cell trait (HbAS), or SCD (HbS), comparing younger versus older adults, and females
versus males. We obtained hematologic parameters in steady-state and hypoxic conditions
and evaluated metabolic markers and cytokines from serum. Kidney function was evaluated
by measuring the urine protein/creatinine ratio and urine osmolality. In vivo studies included von Frey assay, non-invasive plethysmography, and echocardiography.
Histopathological evaluations were performed in lung, liver, spleen, and kidney tissues.
HbS mice displayed elevated hemolysis markers and white blood cell counts, with some
increases more pronounced in older adults. After extended in vivo hypoxia, hemoglobin, platelet counts, and white blood cell counts decreased significantly
in HbS mice, whereas they remained stable in HbA mice. Cytokine analyses showed increased
TNF-alpha in HbS mice. Kidney function assays revealed worsened kidney function in
HbS mice. The von Frey assay showed a lower threshold to response in the HbS mice
than controls, with more noticeable differences in males. Echocardiography in HbS
mice suggested left ventricular hypertrophy and dilatation. Plethysmography suggested
obstructive lung disease and inflammatory changes in HbS mice. Histopathological studies
showed vascular congestion, increased iron deposition, and disruption of normal tissue
architecture in HbS mice. These data correlate with clinical manifestations in SCD
patients and highlight analyses and groups to be included in preclinical therapeutic
studies.
Abbreviations:
ALP (alkaline phosphatase), ALT (alanine aminotransferase), AST (aspartate aminotransferase), BL/6 (C57BL/6), BUN (blood-urea nitrogen), CO (cardiac output), DBIL (direct bilirubin), EDTA (ethylenediaminetetraacetic acid), EDV (end-diastolic volume), EEP (end-expiratory pause), EIP (end-inspiratory pause), ESV (end-systolic volume), GGT (gamma-glutamyl transferase), H&E (hematoxylin and eosin stain), Hb (hemoglobin), HbA (adult hemoglobin), HbF (fetal hemoglobin), HbS (sickle hemoglobin), HR (heart rate), IBIL (indirect bilirubin), IL (interleukin), IVSs (interventricular septum thickness in systole), LDH (lactate dehydrogenase), LLoQ (lower limit of quantification), LV (left ventricle), LVEF (left ventricle ejection fraction), LVFS (left ventricle fractional shortening), LVIDd/s (left ventricle internal diameter in diastole/systole), LVPWd/s (left ventricle posterior wall thickness in diastole/systole), MCV (mean corpuscular volume), MCW (Medical College of Wisconsin), MV (minute ventilation), PEF (peak-expiratory flow), PIF (peak-inspiratory flow), Plt (platelets), Rpef (ratio of time to peak expiratory flow over total expiratory time), QC (quality control), RF (respiratory frequency), SCD (sickle cell disease), TBIL (total bilirubin), Te (time of expiration), Ti (time of inspiration), TNF (tumor necrosis factor), TV (tidal volume), WBC (white blood cell count)To read this article in full you will need to make a payment
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References
- Centers for disease control and prevention.Data & Statistics on Sickle Cell Disease. 2020; (Available at:) (Accessed June 24, 2022)
- Sickle cell disease-genetics, pathophysiology, clinical presentation and treatment.Int J Neonatal Screen. 2019; 5: 20
- Physiological and Aberrant gamma-globin transcription during development.Front Cell Dev Biol. 2021; 9640060
- Sickle cell disease.Ann Intern Med. 2021; 174: ITC1-ITC16
- Knockout-transgenic mouse model of sickle cell disease.Science. 1997; 278: 873-876
- Pathology of Berkeley sickle cell mice: similarities and differences with human sickle cell disease.Blood. 2006; 107: 1651-1658
- Transgenic knockout mice with exclusively human sickle hemoglobin and sickle cell disease.Science. 1997; 278: 876-878
- The transgenic SAD mouse: a model of human sickle cell glomerulopathy.Kidney Int. 1994; 46: 1337-1345
- Towards a transgenic mouse model of sickle cell disease: hemoglobin SAD.EMBO J. 1991; 10: 3157-3165
- Comparative analysis of pain behaviours in humanized mouse models of sickle cell anemia.PLoS One. 2016; 11e0160608
- Human sickle hemoglobin in transgenic mice.Science. 1990; 247: 566-568
- Correction of sickle cell disease by homologous recombination in embryonic stem cells.Blood. 2006; 108: 1183-1188
- Phenotypic characterization the townes sickle mice.Blood. 2014; 124: 4916
- A “shot in the arm" for sickle cell disease.Mol Ther. 2021; 29: 416-417
- Sickle cell disease in mice is associated with sensitization of sensory nerve fibers.Exp Biol Med (Maywood). 2015; 240: 87-98
- Cognitive and behavior deficits in sickle cell mice are associated with profound neuropathologic changes in hippocampus and cerebellum.Neurobiol Dis. 2016; 85: 60-72
- Sleep phenotype in the Townes mouse model of sickle cell disease.Sleep Breath. 2019; 23: 333-339
- Impaired collateral vessel formation in sickle cell disease.Arterioscler Thromb Vasc Biol. 2018; 38: 1125-1133
- The gut microbiome in sickle cell disease: Characterization and potential implications.PLoS One. 2021; 16e0255956
- Testosterone-dependent sex differences in red blood cell hemolysis in storage, stress, and disease.Transfusion. 2016 Oct; 56: 2571-2583
- Sex-based differences in the manifestations and complications of sickle cell disease: Report from the Sickle Cell Disease Implementation Consortium.PLoS One. 2021; 16e0258638
- Sickle cell disease and stroke.Pediatr Neurol. 2019; 95: 34-41
- Disease severity impacts plerixafor-mobilized stem cell collection in patients with sickle cell disease.Blood Adv. 2021; 5: 2403-2411
- Gender-related differences in sickle cell disease in a pediatric cohort: a single-center retrospective study.Front Mol Biosci. 2019; 6: 140
- Sickle cell disease promotes sex-dependent pathological bone loss through enhanced cathepsin proteolytic activity in mice.Blood Adv. 2022; 6: 1381-1393
LABORATORY TJ. Stock No: 013071 | Townes model. 2021 [cited 2021 10/26/2021]; Available at: https://www.jax.org/strain/013071. Accessed October 26, 2021.
Helena Laboratories. Hemoglobin electrophoresis procedure. 2022, Available at: https://www.helena.com/Procedures/Pro15Rev7%20Print.pdf. Accessed April 7, 2022.
- Laboratory animal welfare: Public Health Service policy on humane care and use of laboratory animals by awardee institutions; notice.Fed Regist. 1985; 50: 19584-19585
- Comparison of submental blood collection with the retroorbital and submandibular methods in mice (mus musculus).J Am Assoc Lab Anim Sci. 2016; 55: 570-576
Sysmex C. Sysmex XN-1000™ Hematology Analyzer. 2022 [cited 2022 Feb 7]; Available at: https://www.sysmex.com/US/en/products/hematology/xnseries/pages/xn-1000-hematology-analyzer.aspx. Accessed February 7, 2022.
Analytical Industries, Inc. Handheld oxygen analyzers for medical use (respiratory therapy) AII-2000 and AII-2000 Palm O2. 2022. Available at: https://www.processsensing.com/docs/datasheet/Handheld-Analyzers-for-Medical_V2.pdf. Accessed April 7, 2022.
- Techniques for the detection of sickle cell disease: a review.Micromachines (Basel). 2021; 12
- Relationship between morphologic characteristics of sickle cells and method of deoxygenation.J Lab Clin Med. 1984; 104: 987-994
- Cellular function reinstitution of offspring red blood cells cloned from the sickle cell disease patient blood post CRISPR genome editing.J Hematol Oncol. 2017; 10: 119
- NIH Image to ImageJ: 25 years of image analysis.Nat Methods. 2012; 9: 671-675
- Probability and the westgard rules.Ann Clin Lab Sci. 2003; 33: 113-114
- Optimising the quantification of cytokines present at low concentrations in small human mucosal tissue samples using Luminex assays.J Immunol Methods. 2013; 394: 1-9
- Ways to fit a PK model with some data below the quantification limit.J Pharmacokinet Pharmacodyn. 2001; 28: 481-504
- Efficient analysis of experimental observations.Annu Rev Pharmacol Toxicol. 1980; 20: 441-462
- Quantitative assessment of tactile allodynia in the rat paw.J Neurosci Methods. 1994; 53: 55-63
The Jackson Laboratory Mouse Neurobehavioral Phenotyping Facility (JAX-MNBF). Standard Operating Procedure, Document Number: SOP-JAX-MNBF_SPA Version 1.10, 2018.
- Methods used to evaluate pain behaviors in rodents.Front Mol Neurosci. 2017; 10: 284
Jasmin L, Kohan L, Franssen M, Janni G, Goff JR. The cold plate as a test of nociceptive behaviors: description and application to the study of chronic neuropathic and inflammatory pain models. Pain 199;75(2-3):367-82.
- A simple and inexpensive method for determining cold sensitivity and adaptation in mice.J Vis Exp. 2015;
- Moving to a World Beyond “p < 0.05”.Am Stat. 2019; 73: 1-19
- Hematological parameters in sickle cell anemia patients with and without priapism.Ann Saudi Med. 2006; 26: 439-443
- Original Research: Diametric effects of hypoxia on pathophysiology of sickle cell disease in a murine model.Exp Biol Med (Maywood). 2016; 241: 766-771
- The liver in sickle cell disease.Clin Liver Dis. 2019; 23: 177-189
- Inflammation in sickle cell disease.Clin Hemorheol Microcirc. 2018; 68: 263-299
- Serum IL-6, IL-10, and TNFalpha levels in pediatric sickle cell disease patients during vasoocclusive crisis and steady state condition.Cytokine. 2015; 72: 43-47
- Sickle cell disease: renal manifestations and mechanisms.Nat Rev Nephrol. 2015; 11: 161-171
- The glomerulopathy of sickle cell disease.Am J Hematol. 2014; 89: 907-914
- Renal protection by atorvastatin in a murine model of sickle cell nephropathy.Br J Haematol. 2018; 181: 111-121
- Mouse models for studying pain in sickle disease: effects of strain, age, and acuteness.Br J Haematol. 2012; 156: 535-544
- A noninvasive comparison of left ventricular performance in sickle cell anemia and chronic aortic regurgitation.Clin Cardiol. 1983; 6: 542-548
- Cardiovascular abnormalities in sickle cell disease.J Am Coll Cardiol. 2012; 59: 1123-1133
- Echocardiographic abnormalities in sickle cell disease.Am J Hematol. 2004; 76: 195-198
- Cardiomyopathy in sickle cell disease.Cureus. 2020; 12: e9619
- Sickle cell anemia mice develop a unique cardiomyopathy with restrictive physiology.Proc Natl Acad Sci U S A. 2016; 113: E5182-E5191
- Cardiorespiratory pathogenesis of sickle cell disease in a mouse model.Sci Rep. 2017; 7: 8665
- Lung function in sickle cell disease.Paediatr Respir Rev. 2014; 15: 33-37
- Asthma, allergic sensitization and lung function in sickle cell disease.Allergol Immunopathol (Madr). 2020; 48: 450-457
- The sickle cell mouse lung: proinflammatory and primed for allergic inflammation.Transl Res. 2015; 166: 254-268
- Limitations of a mouse model of sickle cell anemia.Blood Cells Mol Dis. 2002; 28: 146-151
- Spleen histology in children with sickle cell disease and hereditary spherocytosis: hints on the disease pathophysiology.Hum Pathol. 2017; 60: 95-103
- Kidney biopsy findings in children with sickle cell disease: a Midwest Pediatric Nephrology Consortium study.Pediatr Nephrol. 2019; 34: 1435-1445
- How I treat the older adult with sickle cell disease.Blood. 2018; 132: 1750-1760
- Differences between males and females in adult sickle cell pain crisis in eastern Saudi Arabia.Ann Saudi Med. 2004; 24: 179-182
- Practical murine hematopathology: a comparative review and implications for research.Comp Med. 2015; 65: 96-113
- Clinical hematological and biochemical parameters in Swiss, BALB/c, C57BL/6 and B6D2F1 Mus musculus.Animal Model Exp Med. 2020; 3: 304-315
- Inflammatory and oxidative stress phenotypes in transgenic sickle cell mice.Blood Cells Mol Dis. 2016; 62: 13-21
- The platelet count and its implications in sickle cell disease patients admitted for intensive care.Indian J Crit Care Med. 2018; 22: 585-590
- Hypoxia impact on red blood cell-mediated microvascular occlusion and adhesion in sickle cell disease and sickle cell trait.Blood. 2021; 138: 966
- Neutrophils, platelets, and inflammatory pathways at the nexus of sickle cell disease pathophysiology.Blood. 2016; 127: 801-809
- Sickle Cell Disease.Pediatric Respiratory Diseases. 2020; 2020: 529-541
- Pain in sickle cell disease. Rates and risk factors.N Engl J Med. 1991; 325: 11-16
- Alpha and beta thalassemia.Am Fam Physician. 2009; 80: 339-344
- Plasma levels of TNF-alpha in sickle cell patients receiving hydroxyurea.Hematology. 2004; 9: 61-64
- Immunological hallmarks of inflammatory status in vaso-occlusive crisis of sickle cell anemia patients.Front Immunol. 2021; 12559925
- Differential contribution of FXa and thrombin to vascular inflammation in a mouse model of sickle cell disease.Blood. 2014; 123: 1747-1756
- Mouse models of diabetic nephropathy.J Am Soc Nephrol. 2009; 20: 2503-2512
- Development of a chronic kidney disease model in C57BL/6 mice with relevance to human pathology.Nephron Extra. 2013; 3: 12-29
- Assessment of subclinical renal glomerular and tubular dysfunction in children with beta thalassemia major.Children (Basel). 2021; 8
- Hydroxyurea improves nitric oxide bioavailability in humanized sickle cell mice.Am J Physiol Regul Integr Comp Physiol. 2021; 320: R630-RR40
- Hyperfiltration predicts long-term renal outcomes in humanized sickle cell mice.Blood Adv. 2019; 3: 1460-1475
- Combined hydroxyurea and ETA receptor blockade reduces renal injury in the humanized sickle cell mouse.Acta Physiol (Oxf). 2019; 225: e13178
- Long-term endothelin-a receptor antagonism provides robust renal protection in humanized sickle cell disease mice.J Am Soc Nephrol. 2017; 28: 2443-2458
- Sex differences in the trajectory of glomerular filtration rate in pediatric and murine sickle cell anemia.Blood Adv. 2020; 4: 263-265
- Perfluorocarbon emulsion improves oxygen transport of normal and sickle cell human blood in vitro.J Biomed Mater Res A. 2014; 102: 2105-2115
- Echocardiographic assessment of the right heart in mice.J Vis Exp. 2013;
- Echocardiography in Mice.Curr Protoc Mouse Biol. 2011; 1: 71-83
- New metrics for evaluating viral respiratory pathogenesis.PLoS One. 2015; 10e0131451
- Airway inflammation and lung function in sickle cell disease.Pediatr Allergy Immunol Pulmonol. 2019; 32: 92-102
- High protein diet attenuates histopathologic organ damage and vascular leakage in transgenic murine model of sickle cell anemia.Exp Biol Med (Maywood). 2014; 239: 966-974
- How we manage iron overload in sickle cell patients.Br J Haematol. 2017; 177: 703-716
- Iron homeostasis in a mouse model of thalassemia intermedia is altered between adolescence and adulthood.PeerJ. 2020; 8: e8802
- Relationship between Plasma Ferritin Level and Siderocyte Number in Splenectomized beta-Thalassemia/HbE Patients.Anemia. 2012; 2012890471
- Control of Oxidative stress and inflammation in sickle cell disease with the nrf2 activator dimethyl fumarate.Antioxid Redox Signal. 2017; 26: 748-762
- A novel, highly potent and selective phosphodiesterase-9 inhibitor for the treatment of sickle cell disease.Haematologica. 2020; 105: 623-631
- Integrin VLA-4 as a PET imaging biomarker of hyper-adhesion in transgenic sickle mice.Blood Adv. 2020; 4: 4102-4112
- Dietary omega-3 fatty acids protect against vasculopathy in a transgenic mouse model of sickle cell disease.Haematologica. 2015; 100: 870-880
- Sickle cell disease subjects and mouse models have elevated nitrite and cGMP levels in blood compartments.Nitric Oxide. 2020; 94: 79-91
- Transgenic sickle mice have vascular inflammation.Blood. 2003; 101: 3953-3959
- Correction of murine sickle cell disease using gamma-globin lentiviral vectors to mediate high-level expression of fetal hemoglobin.Mol Ther. 2009; 17: 245-252
- Efficacy of homologous inositol hexaphosphate-loaded red blood cells in sickle transgenic mice.Br J Haematol. 2012; 157: 357-369
- Cannabinoid receptor-specific mechanisms to alleviate pain in sickle cell anemia via inhibition of mast cell activation and neurogenic inflammation.Haematologica. 2016; 101: 566-577
- Genetic diminution of circulating prothrombin ameliorates multiorgan pathologies in sickle cell disease mice.Blood. 2015; 126: 1844-1855
- Morphological and functional platelet abnormalities in Berkeley sickle cell mice.Blood Cells Mol Dis. 2008; 41: 109-118
- Base editing of haematopoietic stem cells rescues sickle cell disease in mice.Nature. 2021; 595: 295-302
- A novel human gamma-globin gene vector for genetic correction of sickle cell anemia in a humanized sickle mouse model: critical determinants for successful correction.Blood. 2009; 114: 1174-1185
- Mast cell activation contributes to sickle cell pathobiology and pain in mice.Blood. 2013; 122: 1853-1862
- Bivalent ligand MCC22 potently attenuates nociception in a murine model of sickle cell disease.Pain. 2018; 159: 1382-1391
- Elimination of the fibrinogen integrin alphaMbeta2-binding motif improves renal pathology in mice with sickle cell anemia.Blood Adv. 2019; 3: 1519-1532
- Heme Induces IL-6 and cardiac hypertrophy genes transcripts in sickle cell mice.Front Immunol. 2020; 11: 1910
- Pain-related behaviors and neurochemical alterations in mice expressing sickle hemoglobin: modulation by cannabinoids.Blood. 2010; 116: 456-465
- Tissue factor promotes activation of coagulation and inflammation in a mouse model of sickle cell disease.Blood. 2012; 120: 636-646
- High molecular weight kininogen contributes to early mortality and kidney dysfunction in a mouse model of sickle cell disease.J Thromb Haemost. 2020; 18: 2329-2340
- IL-18 mediates sickle cell cardiomyopathy and ventricular arrhythmias.Blood. 2021; 137: 1208-1218
- Hemolysis in sickle cell mice causes pulmonary hypertension due to global impairment in nitric oxide bioavailability.Blood. 2007; 109: 3088-3098
- Endothelin type A receptors mediate pain in a mouse model of sickle cell disease.Haematologica. 2018; 103: 1124-1135
- Gabapentin alleviates chronic spontaneous pain and acute hypoxia-related pain in a mouse model of sickle cell disease.Br J Haematol. 2019; 187: 246-260
- Heme causes pain in sickle mice via toll-like receptor 4-mediated reactive oxygen species- and endoplasmic reticulum stress-induced glial activation.Antioxid Redox Signal. 2021; 34: 279-293
- Functional MRI BOLD response in sickle mice with hyperalgesia.Blood Cells Mol Dis. 2017; 65: 81-85
- Spatiotemporal alterations in gait in humanized transgenic sickle mice.Front Immunol. 2020; 11561947
- Effect of chronic opioid therapy on pain and survival in a humanized mouse model of sickle cell disease.Blood Adv. 2019; 3: 869-873
- Genetic inactivation of calpain-1 attenuates pain sensitivity in a humanized mouse model of sickle cell disease.Haematologica. 2016; 101: e397-e400
- Sickle cell mice exhibit mechanical allodynia and enhanced responsiveness in light touch cutaneous mechanoreceptors.Mol Pain. 2012; 8: 62
- Transient receptor potential vanilloid 1 mediates pain in mice with severe sickle cell disease.Blood. 2011; 118: 3376-3383
- Cold hypersensitivity increases with age in mice with sickle cell disease.Pain. 2014; 155: 2476-2485
- Endothelin-1 contributes to the progression of renal injury in sickle cell disease via reactive oxygen species.Br J Pharmacol. 2016; 173: 386-395
- Pathologic angiogenesis in the bone marrow of humanized sickle cell mice is reversed by blood transfusion.Blood. 2020; 135: 2071-2084
- Endothelin A receptor antagonist attenuated renal iron accumulation in iron overload heme oxygenase-1 knockout mice.Can J Physiol Pharmacol. 2022; 100: 637-650
- Transgenic sickle mice are markedly sensitive to renal ischemia-reperfusion injury.Am J Pathol. 2005; 166: 963-972
- P-selectin glycoprotein ligand-1 inhibition blocks increased leukocyte-endothelial interactions associated with sickle cell disease in mice.Blood. 2012; 120: 3862-3864
Article info
Publication history
Published online: October 30, 2022
Accepted:
October 23,
2022
Received in revised form:
October 17,
2022
Received:
August 29,
2022
Publication stage
In Press Journal Pre-ProofIdentification
Copyright
Published by Elsevier Inc.
