Restricted accessResearch articleFirst published online 2021-01
Conditioning Regimens in Long-Term Pre-Clinical Studies to Support Development of Ex Vivo Gene Therapy: Review of Nonproliferative and Proliferative Changes
Hematopoietic stem cell gene therapy has become a successful therapeutic strategy for some inherited genetic disorders. Pre-clinical toxicity studies performed to support the human clinical trials using viral-mediated gene transfer and autologous hematopoietic stem and progenitor cell (HSPC) transplantation are complex and the use of mouse models of human diseases makes interpretation of the results challenging. In addition, they rely on the use of conditioning agents that must induce enough myeloablation to allow engraftment of transduced and transplanted HSPC. Busulfan and total body irradiation (TBI) are the most commonly used conditioning regimens in the mouse. Lenticular degeneration and atrophy of reproductive organs are expected histopathological changes. Proliferative and nonproliferative lesions can be observed with different incidence and distribution across strains and mouse models of diseases. The occurrence of these lesions can interfere with the interpretation of pre-clinical toxicity and tumorigenicity studies performed to support the human clinical studies. As such, it is important to be aware of the background incidence of lesions induced by different conditioning regimens. We review the histopathology results from seven long-term studies, five using TBI and two using busulfan.
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References
1.
RichterT, Nestler-ParrS, BabelaR, et al.Rare disease terminology and definitions—a systematic global review: report of the ISPOR Rare Disease Special Interest Group. Value Health, 2015; 18:906–914.
2.
NaldiniL. Genetic engineering of hematopoiesis: current stage of clinical translation and future perspectives. EMBO Mol Med, 2019; 11:e9958.
3.
GinnSL, AmayaAK, AlexanderIE, et al.Gene therapy clinical trials worldwide to 2017: an update. J Gene Med, 2018; 20:1–16.
4.
GalliMC, SerabianM. Regulatory aspects of gene therapy and cell therapy products—A Global Perspective. American Society of Gene & Cell Therapy. New York, NY: Springer International Publishing,, 2015; 1–27.
5.
DownJD, BermanAJ, WarholM, et al.Late tissue-specific toxicity of total body irradiation and busulfan in a murine bone marrow transplant model. Int J Radiat Oncol Biol Phys, 1989; 17:109–116.
6.
Duran-StruuckR, DyskoRC. Principles of bone marrow transplantation (BMT): providing optimal veterinary and husbandry care to irradiated mice in BMT studies. J Am Assoc Lab Anim Sci, 2009; 48:11–22.
7.
NunamakerEA, AndersonRJ, ArtwohlJE, et al.Predictive observation-based endpoint criteria for mice receiving total body irradiation. Comp Med, 2013; 63:313–322.
8.
FasslrinnerF, ScheteligJ, BurchertA, et al.Long-term efficacy of reduced-intensity versus myeloablative conditioning before allogeneic haemopoietic cell transplantation in patients with acute myeloid leukaemia in first complete remission: retrospective follow-up of an open-label, randomised phase 3 trial. Lancet Haematol, 2018; 5:e161–e169.
9.
PeakeK, ManningJ, LewisCA, et al.Busulfan as a myelosuppressive agent for generating stable high-level bone marrow chimerism in mice. J Vis Exp, 2015; e52553.
10.
HaywardAM, LemkeLB, BridgefordEC, et al.Biomethodology and surgical techniques Fox JG, Davisson MT, Quimby FW, Barthold SW, Newcomer CE, Smith AL. (eds.), The Mouse in Biomedical Research. London, UK: Elsevier Inc.,, 2007:482–483.
11.
RambaldiA, GrassiA, MasciulliA, et al.Busulfan plus cyclophosphamide versus busulfan plus fludarabine as a preparative regimen for allogeneic haemopoietic stem-cell transplantation in patients with acute myeloid leukaemia: an open-label, multicentre, randomised, phase 3 trial. Lancet Oncol, 2015; 16:1525–1536.
HurTY, LeeSH, OckSA, et al.Dose-dependent effects of busulfan on dog testes in preparation for spermatogonial stem cell transplantation. Lab Anim Res, 2017; 33:264–269.
14.
KangEM, HsiehMM, MetzgerM, et al.Busulfan pharmacokinetics, toxicity, and low-dose conditioning for autologous transplantation of genetically modified hematopoietic stem cells in the rhesus macaque model. Exp Hematol, 2006; 34:132–139.
15.
Robert-RichardE, GedC, OrtetJ, et al.Human cell engraftment after busulfan or irradiation conditioning of NOD/SCID mice. Haematologica, 2006; 91:1384.
16.
VisigalliI, DelaiS, FerroF, et al.Gene therapy augments the efficacy of hematopoietic cell transplantation and fully corrects mucopolysaccharidosis type I phenotype in the mouse model. Blood, 2010; 116:5130–5139.
17.
LidonniciMR, PaleariY, TiboniF, et al.Multiple integrated non-clinical studies predict the safety of lentivirus-mediated gene therapy for beta-thalassemia. Mol Ther Methods Clin Dev, 2018; 11:9–28.
18.
GibsonFM, AndrewsCM, DiamantiP, et al.A new model of busulphan-induced chronic bone marrow aplasia in the female BALB/c mouse. Int J Exp Pathol, 2003; 84:31–48.
19.
RehgJE, RahijaR, BushD, et al.Immunophenotype of spontaneous hematolymphoid tumors occurring in young and aging female CD-1 mice. [Corrected] [published correction appears in Toxicol Pathol 2015;43:1172]. Toxicol Pathol, 2015; 43:1025–1034.
20.
FrithCH, WardJM, ChandraM. The morphology, immunohistochemistry, and incidence of hematopoietic neoplasms in mice and rats. Toxicol Pathol, 1993; 21:206–218.
21.
RamosMF, BakerJ, AtzpodienEA, et al.Nonproliferative and proliferative lesions of the ratand mouse special sense organs (Ocular [eye and glands], Olfactory and Otic). J Toxicol Pathol, 2018; 31(3Suppl):97S–214S.
22.
DixonD, AlisonR, BachU, et al.Nonproliferative and proliferative lesions of the rat and mouse female reproductive system. J Toxicol Pathol, 2014; 27(3–4Suppl):1S–107S.
23.
CreasyD, BubeA, de RijkE, et al.Proliferative and nonproliferative lesions of the rat and mouse male reproductive system. Toxicol Pathol, 2012; 40(6Suppl):40S–121S.
24.
BishopJB, WassomJS. Toxicological review of busulfan (Myleran). Mutat Res, 1986; 168:15–45.
25.
ChoiYJ, OkDW, KwonDN, et al.Murine male germ cell apoptosis induced by busulfan treatment correlates with loss of c-kit-expression in a Fas/FasL- and p53-independent manner. FEBS Lett, 2004; 575:41–51.
26.
GrimesP, VonsallmannL, FrichetteA. Influence of myleran on cell proliferation in the lens epithelium. Invest Ophthalmol, 1964; 3:566–576.
27.
GutierrezK, GlanznerWG, ChemerisRO, et al.Gonadotoxic effects of busulfan in two strains of mice. Reprod Toxicol, 2016; 59:31–39.
28.
UptonAC, KimballAW, FurthJ. Some delayed effects of atom-bomb radiations in mice. Cancer Res, 1960; 20:1–60.
29.
BuggiaI, LocatelliF, RegazziMB, et al.Busulfan. Ann Pharmacother, 1994; 28:1055–1062.
30.
PaixA, AntoniD, WaissiW, et al.Total body irradiation in allogeneic bone marrow transplantation conditioning regimens: a review. Crit Rev Oncol Hematol, 2018; 123:138–148.
31.
GyurkoczaB, SandmaierBM. Conditioning regimens for hematopoietic cell transplantation: one size does not fit all. Blood, 2014; 124:344–353.
32.
SafwatA, NielsenOS, abd el-BakkyH, et al.Renal damage after total body irradiation in a mouse model for bone marrow transplantation: effect of radiation dose rate. Radiother Oncol, 1995; 34:203–209.
33.
DownJD, BermanAJ, WarholM, et al.Late complications following total-body irradiation and bone marrow rescue in mice: predominance of glomerular nephropathy and hemolytic anemia. Int J Radiat Biol, 1990; 57:551–565.
34.
TravisEL, PetersLJ, McNeillJ, et al.Effect of dose-rate on total body irradiation: lethality and pathologic findings. Radiother Oncol, 1985; 4:341–351.
35.
DownJD, SteelGG. The expression of early and late damage after thoracic irradiation: a comparison between CBA and C57B1 mice. Radiat Res, 1983; 96:603–610.
36.
DownJD. The nature and relevance of late lung pathology following localised irradiation of the thorax in mice and rats. Br J Cancer Suppl, 1986; 7:330–332.
37.
JacksonIL, VujaskovicZ, DownJD. Revisiting strain-related differences in radiation sensitivity of the mouse lung: recognizing and avoiding the confounding effects of pleural effusions. Radiat Res, 2010; 173:10–20.
38.
PetterinoC, NaylorS, MukaratirwaS, et al.Adrenal gland background findings in CD-1 (Crl:CD-1(ICR)BR) mice from 104-week carcinogenicity studies. Toxicol Pathol, 2015; 43:816–824.
39.
SonWC, GopinathC. Early occurrence of spontaneous tumours in CD-1 mice and Sprague–Dawley rats. Toxicol Pathol, 2004; 32:371–374.
40.
BlackwellBN, BucciTJ, HartRW, et al.Longevity, body weight, and neoplasia in ad libitum-fed and diet-restricted C57BL6 mice fed NIH-31 open formula diet. Toxicol Pathol, 1995; 23:570–582.
41.
WardJM. Lymphomas and leukemias in mice. Exp Toxicol Pathol, 2006; 57:377–381.
42.
GenikPC, Bielefeldt-OhmannH, LiuX, et al.Strain background determines lymphoma incidence in Atm knockout mice. Neoplasia, 2014; 16:129–136.
43.
VesselinovitchSD, SimmonsEL, MihailovichN, et al.The effect of age, fractionation, and dose on radiation carcinogenesis in various tissues of mice. Cancer Res, 1971; 31:2133–2142.
44.
RobinE, BermanM, BhoopalamN, et al.Induction of lymphomas in mice by busulfan and chloramphenicol. Cancer Res, 1981; 41(9 Pt 1):3478–3482.
SchiroliG, FerrariS, ConwayA, et al.Preclinical modeling highlights the therapeutic potential of hematopoietic stem cell gene editing for correction of SCID-X1. Sci Transl Med, 2017; 9:eaan0820.
47.
WardJM, RehgJE, MorseHC3rd, et al.Differentiation of rodent immune and hematopoietic system reactive lesions from neoplasias. Toxicol Pathol, 2012; 40:425–434.
48.
Rask-NielsenR, EbbesenP. Reticular neoplasms induced in DBA/2 and CBA mice by intraperitoneal injections of mineral oil. J Natl Cancer Inst, 1965; 35:83–95.
49.
PhilipM, RowleyDA, SchreiberH. Inflammation as a tumor promoter in cancer induction. Semin Cancer Biol, 2004; 14:433–439.
50.
BoormanGA, RaffertyCN, WardJM, et al.Leukemia and lymphoma incidence in rodents exposed to low-frequency magnetic fields. Radiat Res, 2000; 153:627–636.
51.
TriviaiI, ZieglerM, BergholzU, et al.Endogenous retrovirus induces leukemia in a xenograft mouse model for primary myelofibrosis. Proc Natl Acad Sci U S A, 2014; 111:8595–8600.
52.
TillmanH, JankeLJ, FunkA, et al.Morphologic and immunohistochemical characterization of spontaneous lymphoma/leukemia in NSG mice. Vet Pathol, 2020; 57:160–171.
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