Bolder BioPATH, Inc. is a contract pharmacology and pathology company specializing in In Vivo models of Rheumatoid Arthritis, Osteoarthritis, Cancer as well as other autoimmune and inflammation models, Our goals are to provide preclinical (efficacy and toxicity) data to support advancing proteins and small molecules to IND/NDA stage. Since 2003, our mission has been to provide our clients with the highest quality data to move rapidly toward an IND filing. We pride ourselves on being the most flexible contract research lab available. The collaborative relationship between the study sponsor and the contract lab is of the utmost importance and we value supporting our clients’ research objectives. **Recent Publications** - LaBranche T, Bendele A, Omura B, et al. Nerve growth factor inhibition with tanezumab influences weight-bearing and subsequent cartilage damage in the rat medial meniscal tear model. *Ann Rheum Dis*. 2016 July(0:1-8). - Kyostio-Moore S, Piraino S, Berthelette P, et al. Overexpression of cystatin C in synovium does not reduce synovitis or cartilage degradation in established osteoarthritis. *Arthritis Research & Therapy.* 2015;17(1):5. doi:10.1186/s13075-015-0519-3. - Kraus VB, Huebner JL, DeGroot J, Bendele A. The OARSI histopathology initiative – recommendations for histological assessments of osteoarthritis in a guinea pig. *Osteoarthritis Cartilage*. 2010 Oct;18 Suppl 3:S35-52. - Gerwin N, Bendele AM, Glasson S, Carlson CS. The OARSI histopathology initiative – recommendations for histological assessments of osteoarthritis in the rat. *Osteoarthritis Cartilage*. 2010 Oct;18 Suppl 3:S24-34. - Settle S, Vickery L, Nemirovskiy O, Vidmar T, Bendele A, Messing D, Ruminski P, Schnute M, Sunyer T. Cartilage degradation biomarkers predict efficacy of a novel, highly selective matrix metalloproteinase 13 inhibitor in a dog model of osteoarthritis: confirmation by multivariate analysis that modulation of type II collagen and aggrecan degradation peptides parallels pathologic changes. *Arthritis Rheum*. 2010 Oct;62(10):3006-15. - Zimmerman DH, Taylor P, Bendele A, Carambula R, Duzant Y, Lowe V, O’Neill SP, Talor E, Rosenthal KS. CEL-2000: A therapeutic vaccine for rheumatoid arthritis arrests disease development and alters serum cytokine/chemokine patterns in the bovine collagen type II induced arthritis in the DBA mouse model. *Int Immunopharmacol*. 2010 Apr;10(4):412-21. Epub 2010 Jan 13.
Acute DSS-induced colitis in mice and rats
TNBS-induced colitis in mice and rats
Adoptive T-cell transfer-induced colitis in mice
MDR1a Spontaneous Colitis in mice
Anti-CD40-induced Colitis in mice
Acute DSS-induced colitis in mice and rats
TNBS-induced colitis in mice and rats
Adoptive T-cell transfer-induced colitis in mice
MDR1a Spontaneous Colitis in mice
Anti-CD40-induced Colitis in mice
For standard xenograft models, female mice (5 to 6 weeks old) are injected subcutaneously with 0.1 mL of viable human tumor cells on the flank region. All tumor cells are tested... Show more »
For standard xenograft models, female mice (5 to 6 weeks old) are injected subcutaneously with 0.1 mL of viable human tumor cells on the flank region. All tumor cells are tested for infections agents at University of Missouri at Columbia, RADIL lab, and certified clean prior to their use.
Trocar Injection Induction:
Several tumor models, such as the OVCAR-3 human ovarian tumor and the BT474 human mammary tumor, are difficult to consistently grow in vivo when using tissue culture cells. For these models, it is necessary to first generate a few established tumors in severe combined immunodeficient (SCID) mice, or nude mice, and then harvest tumors aseptically, divide into fragment pieces and re-implant those fragments into a cohort of nude mice. For such studies tumor cells obtained from the American Type Culture Collection (ATCC) or other repository are expanded in tissue culture and checked for any infectious agents at the University of Missouri prior to injecting in mice. Once tumors are established, the subcutaneous tumors are removed aseptically and divided in 4 mm3 pieces. These tumor fragments can be cryogenically preserved for future retrieval and use in future studies, or re-implanted immediately. Female mice (5 to 6 weeks old) are implanted subcutaneously with a tumor fragment 4 mm3 of viable tumor on the flank region, using 11 gauge trocar needles.
In these models, tumor growth is monitored and test agent treatment is typically initiated once tumors reach a weight range of 100 to 400 mg.
Mice are visually inspected daily and when visible tumors are evident, tumor size is measured with vernier calipers and tumor volume determined using the formula: Length x Width2/2. When tumor size equals 200 mg +/- 100 mg, the mice are assigned to treatment groups.1-6 End points used to assess drug efficacy include evaluations of tumor growth (comparing tumors in treated versus control mice), tumor cell kill (log10 cell kill), and tumor regression. Regressions are defined as partial if the tumor weight decreases to 50% or less of the tumor weight at the start of treatment without dropping below 63 mg (5 x 5 mm tumor). Both complete regressions (CRs) and tumor-free survivors are defined by instances in which the tumor burden falls below measurable limits (<63 mg).7
These strains of immunocompromised mice vary with regard to the degree of residual immunity and are required to allow the growth of human tumors. Nude mice are the earliest strain of immunocompromised rodents that have been used for growing human tumors and are still the most common strain used. However, some more difficult to grow tumors have been shown to grow in Severe Combined Immunodeficient Mice strains (SCID) where they fail to grow in nude mice. This requires the use of several different strains of immunodeficient mice that have been shown to support the growth of a given tumor line depending on the specific tumor model used.
Bolder BioPATH offers validated models of many inflammatory diseases including psoriasis, dermatitis, delayed type hyper-sensitivity, anaphylaxis, peritonitis, acute edema, pancreatitis and more. Our disease models have been developed over many years, giving us unrivaled expertise. Our inflammatory disease models serve as the... Show more »
Bolder BioPATH offers validated models of many inflammatory diseases including psoriasis, dermatitis, delayed type hyper-sensitivity, anaphylaxis, peritonitis, acute edema, pancreatitis and more. Our disease models have been developed over many years, giving us unrivaled expertise. Our inflammatory disease models serve as the basis for in-depth analysis of compounds at each disease stage, offering answers to your candidates therapeutic effects.
*Inflammatory Bowel... Show more »
Inflammatory Bowel Disease
Inflammatory bowel disease (IBD) is a chronic relapsing inflammation of unknown etiology that presents as two main conditions: Crohn’s disease (CD), a transmural granulomatous inflammation involving segments of the small and large intestine, and ulcerative colitis (UC), which mostly affects the mucosal layer of the large intestine or colon1.
Historically, most animal models of IBD have been chemically induced using agents such as dextran sulfate sodium (DSS), 2,4,6-Trinitrobenzenesulfonic acid (TNBS) or oxazolone. Of these, the DSS model, which initiates a lethal inflammatory bowel disease that resembles human ulcerative colitis, is one of the most commonly used. More recently, models of mouse IBD have been developed using naturally occurring mutant mouse strains (e.g. C3H/HeBir), gene-knockout, and transgenic strains that spontaneously develop colitis. Additionally, intestinal inflammation with characteristics resembling both UC and CD has been shown to occur in immunodeficient mice that are reconstituted with the CD45RBhigh subset of CD4+ T cells1. The adoptive T-cell transfer model has proven to be a highly reproducible and easily manipulated model of colitis2.
The mechanisms underlying the development of IBD are not well understood. Traditionally, it was believed to be an infectious disease; however, the failure to find a causative microbe and the effectiveness of steroid therapy led to the view that IBD may be an immunological or autoimmune disorder. Recent research has suggested that genetic susceptibility and an emerging mucosal immune response against gut constituents are important factors, and that T cells and tumor necrosis factor alpha (TNF-a) play crucial roles in the pathogenesis of the disease1.
Although animal models of IBD are not perfect analogs of the human disease, mouse models have characteristics that closely resemble both UC and CD in humans. The DSS model has similarities to UC and is characterized by colonic epithelial cell lesions and acute inflammation followed by chronic colitis once DSS administration has been terminated. Clinical symptoms include diarrhea and bloody stool1. The CD45RBhighadoptive T-cell transfer model has characteristics of UC (like, a diffuse distribution of lesions in the large intestine, crypt elongation and branching, and extensive mucin depletion) as well as CD (like, transmural inflammation, and the presence of many macrophages and lymphocytes with the occasional multinucleated giant cell). Clinical symptoms associated with the adoptive T-cell transfer model include progressive weight loss and loose stool with mucus, but not diarrhea or bloody stool2.
Conventional treatments for IBD include corticosteroids and 5-aminosalicylic acid derivatives, which are not effective in inducing clinical remission and which can have deleterious side effects. Antibody therapies, including anti-TNF-a, anti-IL-12, and anti-IFN-g antibody therapies, have been shown to be effective in the treatment of IBD although repeated systemic antibody administration may also pose potential complications. Novel approaches using gene therapy have also shown some success.
Animal models of osteoarthritis (OA) that are commonly used to study the pathogenesis of cartilage degeneration and potential therapy can be either naturally occurring or induced. Induction involves surgical manipulation or injection of matrix modifying agents into the joint. The knee joint is the most common joint that is affected (spontaneous OA) or utilized (induced). Spontaneous OA occurs reliably only in certain animal species, but some form of surgical or chemical OA can be induced in any animal species. By using different animals, one can achieve diversity in the lesion characteristics. Alternatively, diverse lesions of varying severity can be obtained by applying different injuries to animals of the same species. This would then permit investigation of different types of therapeutic interventions. Recent studies that have compared profiles of cartilage degradation in various models with those in human or spontaneous animal disease indicate that most OA models have more similarities than differences (2, 3, 4, 5). Model selection for a particular purpose generally comes down to selection of the desired animal species and, within that species, having the desired morphologic and biochemical changes that will allow appropriate evaluation of the efficacy of a potential therapeutic agent in a reasonable period of time.
The diversity of models and species often leaves the investigator confused about the choice of an appropriate model, yet, as a result of preliminary work, the investigator may already have chosen a particular animal species. A further important consideration relates to the morphologic features of the lesion and to knowledge about mediators involved in the pathogenesis, particularly when a pharmaceutic agent is being tested. For example, inhibitors of collagenase should be tested in models where collagen degradation is of sufficient severity to generate an observable morphologic change. Aggrecanase inhibitors, on the other hand, should be tested in models where the primary morphologic change is proteoglycan loss, so that collagen loss does not obscure the proteoglycan evaluation. Another way to address this issue within most instability models is by individual evaluation of load-bearing zones of articular cartilage. In virtually all surgical models, there are areas of mechanical abrasion adjacent to areas of milder, enzymatic degradation. Histopathologic evaluation that takes into account these zonal variabilities will often be as useful as utilizing multiple models of different severity. However, just as criteria for efficacy are set at the onset of a human clinical trial, animal studies must also have pre-defined parameters. Criteria for success should depend on the potential limits of the agent under investigation.
Animal models of rheumatoid arthritis (RA) with a proven track record of predictability... Show more »
Animal models of rheumatoid arthritis (RA) with a proven track record of predictability for efficacy in humans include: rat adjuvant arthritis (AIA) , rat type II collagen arthritis (CIA), mouse type II collagen arthritis (CIA) and antigen- induced arthritis in several species. Agents currently in clinical use (or trials) that are active in these models include: corticosteroids, methotrexate, nonsteroidal anti-inflammatory drugs, cyclosporin A, leflunomide interleukin-1 receptor antagonist (IL-1ra) and soluble TNF receptors. For some of these agents, the models also predict that toxicities seen at higher doses for prolonged dosing periods would preclude dosing in humans at levels that might provide disease modifying effects.
In comparison to the osteoarthritis models, RA models are relatively easy to perform, have good reproducibility of data and are generally of short duration. Most of the RA models have some pathological features that are similar to those occurring in human disease. Important differences include 1) animal models of RA progress much more rapidly than does human disease and thus are characterized by primarily acute inflammatory responses and 2) rodents have a tendency to have marked bone resorption and bone formation (especially periosteal/ endosteal) in response to joint inflammation.
Ultimate selection of an animal model for studies on pathogenesis or effects of inhibitors of RA requires consideration of the purpose of the study. If the need is for rapid generation of preclinical efficacy/toxicity data to facilitate entry into clinical trials, selection of one of the induced (AIA, CIA) models is probably most appropriate. Generation of efficacy data in one of these models is procedurally straightforward and therefore should be reproducible. Test animals are readily available should the need for large-scale testing emerge. In addition, these models (AIA, CIA) have excellent track records for predicting activity and toxicity (at high doses of various agents) in humans. So comparative studies between older vs newer anti-arthritics can be done. Also, since these models are highly reproducible, examination of structure activity relationships between various molecules should be easily achievable.
Activity of commonly used small molecule anti-arthritic agents such as dexamethasone, indomethacin (and other NSAIDs including cyclo-oxygenase 2 inhibitors) and methotrexate (adjuvant only) are predicted by the developing rat adjuvant and established rat type II collagen arthritis models.
Dexamethasone and other corticosteroids are used in the clinical treatment of RA but only at low doses because of the toxicities associated with chronic use. Both animal models predict that corticosteroids have the potential to beneficially affect all aspects of RA and that they have the potential to be disease modifying but that toxicities associated with chronic dosing preclude their use at these efficacious doses. So the models predict that only modest clinical responses could be expected with a non-toxic dosing regime.
Systemic lupus erythematosus (SLE) is a multifactorial autoimmune disease characterized by polysystemic inflammation as a result of auto-reactive T and B cells. In mammals, the immune system can... Show more »
Systemic lupus erythematosus (SLE) is a multifactorial autoimmune disease characterized by polysystemic inflammation as a result of auto-reactive T and B cells. In mammals, the immune system can sustain prolonged adaptive immune responses to nearly any foreign antigen by generating, through genetic recombination, lymphocytes with highly diverse receptors. Due to the randomness of this process, a proportion of T and B cells inevitably have receptors that are directed against self-antigen; however, these lymphocytes are typically eliminated, reprogrammed, or inactivated in the lymphoid organs. In SLE patients, regulatory T cells that clear out autoantibody-antigen complexes may be reduced or lacking, and the continued presence of these complexes activate an autoimmune positive feedback loop that allows the disease to progress beyond the inciting autoantigen through “epitope spreading”1.
SLE has been recognized in a variety of species including humans, mice, rats, and other domesticated animals. SLE in mice closely resembles SLE in humans, including autoantibody production and renal disease. Mouse models of SLE fall under two main categories—spontaneous and induced—and each model presents its own iterations of lupus-like disease with a subset of symptoms similar to those seen in human SLE2.The most commonly used mouse models are the NZBWF1 and MRL/lpr strains, which develop spontaneous disease characterized by hyperactive B and T cells, high titers of several autoantibodies directed against nuclear antigens, defective clearance of immune complexes, and fatal immune glomerulonephritis1.
Various factors from genetic defects to infectious agents and drug exposure contribute to the pathogenesis of SLE. Patients with SLE have been shown to have defective B- and T-cell tolerance to nuclear antigens, hypomethylated DNA, an increased rate of apoptosis, and defective clearance of apoptotic debris; it has been hypothesized that, in SLE patients, apoptotic debris becomes immunogenic through abnormal processing. Plasmacytoid and myeloid dendritic cells and B-cell activating factor (BAFF) have been implicated in the disease’s feedback loop.
Clinical symptoms of SLE differ depending upon the underlying genetic abnormalities and the anatomic location of the cellular responses against self-antigen. Autoantibody-antigen immune complexes contribute to the clinical manifestation of the disease by causing lesions in blood vessels and tissues where they accumulate. In the case of lupus nephritis, circulating immune complexes deposit in the glomerular subendothelial space and in the mesangium, leading to deterioration of the glomerulus and eventual end-stage renal disease3. In addition to renal disease (glomerulonephritis, interstitial nephritis, vasculitis, and proteinuria), SLE patiens may present with swollen joints, skin rash, hematologic disorders, and respiratory and neurologic dysfunction1. Although mouse models are primarily characterized by the development of nephritis, some other lupus-like symptoms may develop in certain strains2.
Over the years, a variety of therapeutic approaches for disease intervention have been taken. Treatments have included generalized immunosuppressants (e.g. methotrexate), specific cytokine blocking (e.g. INFa), innate immune inhibitors (e.g. chloroquine), adaptive immune inhibitors (e.g. monoclonal antibodies against BAFF), and costimulation inhibitors (e.g. abatacept). Generalized immunosuppressants are considered undesirable because they increase a patient’s susceptibility to bacterial or fungal infections, so recent research has focused on the more specific approaches with a monoclonal antibody against BAFF (anti-BLys; belimumab) showing promising results1.
"Bolder BioPATH team was very response to our needs for reading our histology slides. They also provided suggestions on how to better prepare our samples."
Bolder BioPATH, Inc. has not received any endorsements.