Abstract
Genetically defined mice have been the cornerstone of cancer research and resources at The Jackson Laboratory since the founding of the laboratory nearly nine decades ago. The Jackson ...Laboratory distributes more than 10,500 strains to the scientific community, many with applications in cancer research, including mouse models for specific cancers, xenograft models, immunodeficient platforms for PDX studies, multi-purpose “tool strains” (such as CRISPR cas9-expressing lines), recombinase expressing strains, as well as conditional and inducible expression lines (such as Cre-lox). New to the JAX Repository collection is a KrasLSL-A146T strain with inducible mutant Kras A146T for use in research of gastrointestinal tract and hematological cancers. The tamoxifen-inducible model for pancreatic ductal adenocarcinoma (PDAC), KPCX, and the related conditional activatable Kras allele/floxed Trp53 double mutant line, KP, have recently been developed. Inducible expression of mutant human speckle-type POZ protein (SPOP) in one newly acquired line allows study of prostate tumorigenesis. The portfolio of strains useful for engraftment studies has expanded, complementing the versatile humanized NOD scid gamma (NSG) platform lines. For custom generation of mutant strains, the Repository distributes a variety of research tool strains such as cas9-expressing lines, both constitutive expressing and Cre-inducible models, on different defined genetic backgrounds. Other recently added mutants allow research of the role of inflammation in cancer and screening immunomodulatory cancer therapies.
In addition to safeguarding each strain by cryopreservation, the Repository‘s comprehensive quality control program confirms mutation identity and genetic background, and screens for unwanted alleles (such as stray GFP, cre, lacZ, etc.). Investigators can search for specific strains on the JAX Mice website, or peruse the Oncology Therapeutic Area webpage for cancer research related strains and resources. Researchers can submit their strains to be considered for inclusion in the Repository on The Jackson Laboratory website:
www.jax.org/donate-a-mouse
The Jackson Laboratory Repository is supported by the NIH, The Howard Hughes Medical Institute, and other private charitable foundations.
Citation Format: Deborah Boswell, Stephen Rockwood, Cathleen Lutz, The JAX Repository Team. Mouse strains for cancer research at The Jackson Laboratory Repository abstract. In: Proceedings of the American Association for Cancer Research Annual Meeting 2019; 2019 Mar 29-Apr 3; Atlanta, GA. Philadelphia (PA): AACR; Cancer Res 2019;79(13 Suppl):Abstract nr 4625.
The “housekeeping” sodium/hydrogen exchanger, NHE1, mediates the electroneutral 1:1 exchange of Na
+ and H
+ across the plasma membrane. NHE1 is ubiquitous and is studied extensively for regulation ...of pH
i
, cell volume, and response to growth factors. We describe a spontaneous mouse mutant,
s low-
w ave
e pilepsy, (
swe), with a neurological syndrome including ataxia and a unique epilepsy phenotype consisting of 3/sec absence and tonic-clonic seizures.
swe was fine-mapped on Chromosome 4 and identified as a null allele of
Nhe1. Mutants show selective neuronal death in the cerebellum and brainstem but otherwise are healthy. This first example of a disease-causing mutation in an
Nhe gene provides a new tool for studying the delicate balance of neuroexcitability and cell survival within the CNS.
Proximal spinal muscular atrophy (SMA) is the leading genetic cause of infant mortality. Traditionally, SMA has been described as a motor neuron disease; however, there is a growing body of evidence ...that arrhythmia and/or cardiomyopathy may present in SMA patients at an increased frequency. Here, we ask whether SMA model mice possess such phenotypes. We find SMA mice suffer from severe bradyarrhythmia characterized by progressive heart block and impaired ventricular depolarization. Echocardiography further confirms functional cardiac deficits in SMA mice. Additional investigations show evidence of both sympathetic innervation defects and dilated cardiomyopathy at late stages of disease. Based upon these data, we propose a model in which decreased sympathetic innervation causes autonomic imbalance. Such imbalance would be characterized by a relative increase in the level of vagal tone controlling heart rate, which is consistent with bradyarrhythmia and progressive heart block. Finally, treatment with the histone deacetylase inhibitor trichostatin A, a drug known to benefit phenotypes of SMA model mice, produces prolonged maturation of the SMA heartbeat and an increase in cardiac size. Treated mice maintain measures of motor function throughout extended survival though they ultimately reach death endpoints in association with a progression of bradyarrhythmia. These data represent the novel identification of cardiac arrhythmia as an early and progressive feature of murine SMA while providing several new, quantitative indices of mouse health. Together with clinical cases that report similar symptoms, this reveals a new area of investigation that will be important to address as we move SMA therapeutics towards clinical success.
Mutations of the alpha1A calcium channel subunit have been shown to cause such human neurological diseases as familial hemiplegic migraine, episodic ataxia-2, and spinocerebellar ataxia 6 and also to ...cause the murine neurological phenotypes of tottering and leaner. The leaner phenotype is recessive and characterized by ataxia with cortical spike and wave discharges (similar to absence epilepsy in humans) and a gradual degeneration of cerebellar Purkinje and granule cells. The mutation responsible is a single-base substitution that produces truncation of the normal open reading frame beyond repeat IV and expression of a novel C-terminal sequence. Here, we have used whole-cell recordings to determine whether the leaner mutation alters calcium channel currents in cerebellar Purkinje cells, both because these cells are profoundly affected in leaner mice and because they normally express high levels of alpha1A. In Purkinje cells from normal mice, 82% of the whole-cell current was blocked by 100 nM omega-agatoxin-IVA. In Purkinje cells from homozygous leaner mice, this omega-agatoxin-IVA-sensitive current was 65% smaller than in control cells. Although attenuated, the omega-agatoxin-IVA-sensitive current in homozygous leaner cells had properties indistinguishable from that of normal Purkinje neurons. Additionally, the omega-agatoxin-IVA-insensitive current was unaffected in homozygous leaner mice. Thus, the leaner mutation selectively reduces P-type currents in Purkinje cells, and the alpha1A subunit and P-type current appear to be essential for normal cerebellar function.
Abstract
Systemic Lupus Erythematosus is a chronic autoimmune disease that causes systemic damage throughout the body and is characterized by a broad spectrum of clinical manifestations. The ...pathogenic mechanisms underlying the disease remain unclear, but are thought to be the result of gene-by-environment interactions that lead to the breakdown of self-tolerance in susceptible individuals. Mouse models of SLE have been instrumental in our understanding of the genetic and cellular requirements for SLE induction, and represent a valuable resource for testing SLE therapies and drug candidates in vivo. No one mouse model completely recapitulates the complexity of the human disease, thus it is crucial to evaluate the mode of action and therapeutic efficacy of targeted therapeutics in multiple SLE mouse models. Here we demonstrate an effective program for screening therapeutic candidates using two well-established SLE mouse models, NZBWF1/J mice and MRL/MpJ-Faslpr/J mice, which mimic the genetic and pathophysiological aspects of the human disease. We characterize the disease course in each model in order to optimize the treatment schedule for either prophylactic or therapeutics drug efficacy studies. Using the immunosuppressant drug cyclophosphamide, we longitudinally compare the clinical measures of the disease to immune cell profiles during both in-life and at study terminus. This program provides a valuable resource for the investigation into the specific effects novel therapeutics have on immune homeostasis.
Abstract
The Jackson Laboratory Repository (JAX) serves as a centralized facility for the development, distribution and cryopreservation of mouse models of human biology and disease. JAX distributes ...more than 10,500 strains to the scientific community, many with applications in immunology research, including mouse models for autoimmune disease, infectious disease, immunodeficient platforms for PDX studies, and multi-purpose “tool strains” (such as CRISPR cas9-expressing lines), recombinase expressing strains, as well as conditional and inducible expression lines.
One of our newest strains is a conditional knock-out (KO) of STING/MYPS, a pathogen sensor involved in the activation of IL6 and IFNb. Additions to our extensive set of interleukin alleles include a SMART-17A knock-in (KI) expressing human NGFR on IL17A expressing cells and a KI/KO of Il9-expressing iCre/EYFP. The latest reporter and cre-expressing strains include an inducible cre KO/KI to Lyz2, and a KI/KO strain with dual reporters of chemokine Ccr2RFP and Cx3cr1GFPactivity, and two strains expressing the INDIA apoptosis reporter, one with widespread conditional expression and the other with expression in germinal center B cells. KOMP mice, offered via the MMRRC portion of the JAX Mouse Repository, provide a growing number of targeted immunological mutations.
Donating a strain to the Repository fulfills NIH’s requirements for sharing mice. Researchers wishing to have mice considered for inclusion in the Repository are encouraged to submit strains: www.jax.org/donate-a-mouse.
This work is supported by NIH, HHMI and private foundations.
Abstract
The Jackson Laboratory Mouse Repository continues to expand its diverse collection of mutant mouse strains, many of which have preclinical applications related to human cancer. Over 11,000 ...Repository mouse lines are accessible to the scientific community, each described and easily found on the JAX website. Among the hundreds of new strains made available annually by the Repository are mice from NIH-funded mouse projects and programs (SMSR, KOMP, MMRRC). A subset of strains relevant to cancer research is highlighted here. A number of mouse lines have been designed to be models for studying specific types of cancers. Recent additions include a model for small cell lung cancer carrying floxed alleles of Rb1 and Trp53 and a Cre recombinase regulatable Myc allele; a conditional strain, in which exon 15 of Apc is floxed, useful for generating models of ovarian and colorectal cancer; and a double floxed Wwtr1 Yap1 line with applications in studies of the role of the Hippo signaling pathway in tumorigenesis and tumor suppression. The set of engraftment models continues to grow. Typically immunodeficient NSG-based (but not exclusively) strains allow xenotransplantation of human tissues, cells or tumors. The set of research tool strains encompassing Cre-lox, Tet-On/-Off and CRISPR cas9 technologies also continues to grow as does the set of reporter strains (fluorescent protein, luciferase, lacZ, etc.), enabling researchers to customize models for particular applications. A transgenic strain expressing Cre recombinase in gastric parietal cells has applications in gastric cancer research. A reporter line that expresses JAG1 and luciferase in Cre-expressing tissues can be used to study prostate cancer. Other resources enabling cancer research at JAX include the JAX patient-derived xenograft (PDX) cancer model resource offering PDX tumor-bearing NSG mice as a preclinical platform for efficacy studies. The Onco-Hu model platform can be used for immunotherapy engraftment studies. The Oncology Therapeutic Area web page is tailored to showcase JAX strains and resources related to cancer research. Repository strains are tested in our comprehensive quality control program to confirm expected mutation identity and genetic background, as well as to screen for the presence of contaminating alleles (GFP, cre, lacZ, etc.). Mice harboring unexpected, often overlooked, genetically engineered alleles can dramatically confound reproducibility between labs. In addition, the JAX Reproductive Sciences group performs cryopreservation to safeguard each strain and can rapidly create cohorts for studies using IVF technology. Researchers wishing to have strains considered for inclusion in the Repository can submit them on the JAX website at www.jax.org/donate-a-mouse. The Jackson Laboratory Repository is supported by the NIH, The Howard Hughes Medical Institute, and several private charitable foundations.
Citation Format: Deborah Boswell, Stephen Rockwood, Cathleen Lutz, The JAX Repository Team. The Jackson Laboratory Repository: Mouse strains relevant to human cancer abstract. In: Proceedings of the American Association for Cancer Research Annual Meeting 2018; 2018 Apr 14-18; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2018;78(13 Suppl):Abstract nr 5094.
Genetic background significantly affects phenotype in multiple mouse models of human diseases, including muscular dystrophy. This phenotypic variability is partly attributed to genetic modifiers that ...regulate the disease process. Studies have demonstrated that introduction of the γ-sarcoglycan-null allele onto the DBA/2J background confers a more severe muscular dystrophy phenotype than the original strain, demonstrating the presence of genetic modifier loci in the DBA/2J background. To characterize the phenotype of dystrophin deficiency on the DBA/2J background, we created and phenotyped DBA/2J-congenic Dmdmdx mice (D2-mdx) and compared them with the original, C57BL/10ScSn-Dmdmdx (B10-mdx) model. These strains were compared with their respective control strains at multiple time points between 6 and 52 weeks of age. Skeletal and cardiac muscle function, inflammation, regeneration, histology and biochemistry were characterized. We found that D2-mdx mice showed significantly reduced skeletal muscle function as early as 7 weeks and reduced cardiac function by 28 weeks, suggesting that the disease phenotype is more severe than in B10-mdx mice. In addition, D2-mdx mice showed fewer central myonuclei and increased calcifications in the skeletal muscle, heart and diaphragm at 7 weeks, suggesting that their pathology is different from the B10-mdx mice. The new D2-mdx model with an earlier onset and more pronounced dystrophy phenotype may be useful for evaluating therapies that target cardiac and skeletal muscle function in dystrophin-deficient mice. Our data align the D2-mdx with Duchenne muscular dystrophy patients with the LTBP4 genetic modifier, making it one of the few instances of cross-species genetic modifiers of monogenic traits.
Abstract
The mouse continues to provide researchers a means by which to address complicated immunological questions. The emergence of disruptive genetic engineering technologies have proven to be ...very applicable to the mouse, spawning a new generation of promising preclinical models. To facilitate access to this flourishing resource, The Jackson Laboratory (JAX) Mouse Repository offers one of the largest sets of well-characterized, genetically engineered mutant mice. Hundreds of new strains are added annually to the collection.
Our newest strains include conditionally targeted interleukins, often combined with Cre recombinase and/or reporters. The conditional Il33 knock-out/in (KO/KI) carries an EGFP reporter, a KO/KI to the Il5 gene results in Il15-specific expression of tdTomato and humanized cre, a similar KO/KI to the Il22 gene results in tdTomato/cre expression in intestinal lymphoid tissues, and a KI to Il21 results in expression of Verde green (VFP). Another new strain expresses cre and GFP within peripheral CD8+ T cells.
MMRRC program mice include a suite of strains expressing and responding to influenza virus hemagglutinin (HA) neo-self antigen for the study of immune regulation, tolerance and autoimmunity. A Pad4 KO, exhibits absent histone deimination in neutrophils and a defective response to acute infection. KOMP mice, also offered via the MMRRC portion of the JAX Mouse Repository, provide a growing number of targeted immunological mutations. Donating a strain to the Repository fulfills NIH’s requirements for sharing mice. Researchers wishing to have mice considered for inclusion in the Repository are encouraged to submit strains: www.jax.org/donate-a-mouse.
This work is supported by NIH, HHMI and private foundations.
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