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Use of stem cell derived peripheral neurons as a model of chemotherapeutic induced peripheral neuropathy
376
Abstracts
0135 Use of stem cell derived peripheral neurons as a model of chemotherapeutic induced peripheral neuropathy Shannon Delaneya, Claudia Winga, Nicole Favrea, Greg Luermanb, Eileen Dolana a
University of Chicago, Chicago, IL, USA Axiogenesis AG, Köln, Germany
b
Chemotherapy induced peripheral neuropathy (CIPN) is one of the most common and potentially permanent side effects of chemotherapy, second only in frequency to hematopoietic toxicity. About 20–40% of cancer patients with estimates of about 400,000 patients each year develop CIPN as a consequence of treatment with platinum analogues (cisplatin, oxaliplatin, carboplatin), taxanes (paclitaxel, docetaxel), vinca alkaloids (vincristine), thalidomide, epothilones and bortezomib. Moreover, CIPN can lead to dose reduction of the chemotherapeutic agent or possible cessation of treatment. This may have an adverse impact on cancer treatment and disease outcomes. In this study, we evaluate the utility of induced pluripotent stem cell-derived human peripheral neurons as a cellular model to assist in the development of neuroprotective agents and validate genes thought to contribute to CIPN. Upon treatment of human peripheral neurons with paclitaxel, vincristine, bortezomib and cisplatin (1 nM to 10 μM) up to 72 h, we measured morphological characteristics such as relative outgrowth, process length, branches, and outgrowth intensity as well as cell health as measured by Cell titer Glo. We identified reproducible decreases in relative neurite length per magnitude of drug concentration. The morphological changes differed across drugs, with vincristine and bortezomib treatment showing the most dramatic effect on neurite outgrowth, paclitaxel showing an intermediate effect, and cisplatin showing no effect up to 10 μM of drug. These cells provide a novel and innovative means to evaluate neurotoxic chemotherapeutic agents and identify new drugs to overcome this devastating side effect of chemotherapy. doi:10.1016/j.vascn.2016.02.134
0136 Enhance in pentylenetetrazole or maximal electro shock induced seizures in experimental models of ulcerative colitis: An interplay of altered cytokines level, adhesion molecules, blood brain barrier disruption Bikash Medhi, Rakesh K. Sewal, Manish Modi, Uma Nahar Saikia, Amitava Chakrabarti PGIMER, Chandigarh, India Aim of study was designed to assess the seizure susceptibility in experimental models of ulcerative colitis. Experimental colitis was induced by single intra-colonic instillation of 20 mg trinitrobenzene sulphonic acid (TNBS) in wistar rats. Zonisamide (ZNS), and thalidomide (Thali) were given to two different groups of animals along with TNBS instillation. Two groups of animals were subjected to administration of vehicle and TNBS in colon respectively without any treatment. After 14 days, seizures were induced either by pentylenetetrazole (PTZ) or maximal electroshock (MES). Seizure parameters, colitis score, oxidative stress and TNF-α, IL-6, IL-1β, ICAM-1, ICAM-2, VCAM-1, MMP-9 level in brain and serum samples were assessed. Histopathological examination and blood brain barrier permeability study was done.
Mean colitis score decreased in thalidomide treated group but not in ZNS treated group. Seizures were decreased in ZNS and Thali treated groups. Seizures were significantly higher in TNBS instilled group as compared to veh group. Oxidative stress was significantly lower in ZNS and Thali treated groups as compared to TNBS instilled group. TNF-α, IL-6, IL-1β, ICAM-1, VCAM-1, MMP-9 levels were found increased in TNBS instilled animals treated with vehicle whereas a reverse observation was noted for ICAM-2 level in brain and serum. Histopathological showed establishment of ulcerative colitis model in animals. Blood brain barrier was found distorted in TNBS instilled groups of animals. Seizure susceptibility may increase in experimental models of ulcerative colitis and anti-inflammatory agent may ameliorate this effect. doi:10.1016/j.vascn.2016.02.135
0137 Utility of an in vitro platform to assess neuronal toxicity using human iPSC-derived neurons Dinah Misnera, Chelsea Snyderc, Tin Ngob, Lanlan Yua, Nicola Stagga, Karin Staflina a
Genentech, South San Francisco, CA, USA University of California, Davis, Davis, CA, USA c Gilead Sciences, Foster City, CA, USA b
A major cause of drug attrition from clinical trials is neurotoxicity, a problem likely due to the lack of predictivity of currently available preclinical models to human safety. One of the most common druginduced neurotoxicities observed clinically is peripheral neuropathy. Existing in vivo animal models are time-consuming, expensive, and lack a direct species relationship to humans. Current in vitro models consist of immortalized cell lines and primary rodent cultures, which show variable correlation to human physiology. There is an unmet need for higher throughput, in vitro human-based assays that can identify compounds with neurotoxic and neuropathic potential. These in vitro assays can precede in vivo animal studies, and augment with data in the most relevant species before clinical trials begin. We chose to establish such assays with human induced pluripotent stem cells (iPSCs) that have been differentiated into mature neurons. These iPSC neurons offer a renewable source of mature cells that may be used to investigate neurotoxic potential. Our characterization of these neurons shows that they express mature markers at both transcript and protein levels. Using kinetic, high-throughput imaging we evaluated a large panel of test compounds known to cause peripheral neuropathy in the clinic and examined effects on neurite outgrowth and viability. Results from our assays show that these test compounds exert neurotoxicity in a classspecific manner and that our assessment correlates to clinical findings. We propose the use of this in vitro human assay to evaluate the potential for neurotoxicity and to provide utility in drug safety applications. doi:10.1016/j.vascn.2016.02.136
0138 NeuroSafe: A human integrated in vitro Neurotoxicity Safety Platform Loredana Redaellia, Giovanna Marescaa, Sara Tremoladaa, Christina Kuhna, Matteo Brioschia, Dietmar Hessb, Elke Guentherb, Lia Scarabottoloa
Abstracts
0135 Use of stem cell derived peripheral neurons as a model of chemotherapeutic induced peripheral neuropathy Shannon Delaneya, Claudia Winga, Nicole Favrea, Greg Luermanb, Eileen Dolana a
University of Chicago, Chicago, IL, USA Axiogenesis AG, Köln, Germany
b
Chemotherapy induced peripheral neuropathy (CIPN) is one of the most common and potentially permanent side effects of chemotherapy, second only in frequency to hematopoietic toxicity. About 20–40% of cancer patients with estimates of about 400,000 patients each year develop CIPN as a consequence of treatment with platinum analogues (cisplatin, oxaliplatin, carboplatin), taxanes (paclitaxel, docetaxel), vinca alkaloids (vincristine), thalidomide, epothilones and bortezomib. Moreover, CIPN can lead to dose reduction of the chemotherapeutic agent or possible cessation of treatment. This may have an adverse impact on cancer treatment and disease outcomes. In this study, we evaluate the utility of induced pluripotent stem cell-derived human peripheral neurons as a cellular model to assist in the development of neuroprotective agents and validate genes thought to contribute to CIPN. Upon treatment of human peripheral neurons with paclitaxel, vincristine, bortezomib and cisplatin (1 nM to 10 μM) up to 72 h, we measured morphological characteristics such as relative outgrowth, process length, branches, and outgrowth intensity as well as cell health as measured by Cell titer Glo. We identified reproducible decreases in relative neurite length per magnitude of drug concentration. The morphological changes differed across drugs, with vincristine and bortezomib treatment showing the most dramatic effect on neurite outgrowth, paclitaxel showing an intermediate effect, and cisplatin showing no effect up to 10 μM of drug. These cells provide a novel and innovative means to evaluate neurotoxic chemotherapeutic agents and identify new drugs to overcome this devastating side effect of chemotherapy. doi:10.1016/j.vascn.2016.02.134
0136 Enhance in pentylenetetrazole or maximal electro shock induced seizures in experimental models of ulcerative colitis: An interplay of altered cytokines level, adhesion molecules, blood brain barrier disruption Bikash Medhi, Rakesh K. Sewal, Manish Modi, Uma Nahar Saikia, Amitava Chakrabarti PGIMER, Chandigarh, India Aim of study was designed to assess the seizure susceptibility in experimental models of ulcerative colitis. Experimental colitis was induced by single intra-colonic instillation of 20 mg trinitrobenzene sulphonic acid (TNBS) in wistar rats. Zonisamide (ZNS), and thalidomide (Thali) were given to two different groups of animals along with TNBS instillation. Two groups of animals were subjected to administration of vehicle and TNBS in colon respectively without any treatment. After 14 days, seizures were induced either by pentylenetetrazole (PTZ) or maximal electroshock (MES). Seizure parameters, colitis score, oxidative stress and TNF-α, IL-6, IL-1β, ICAM-1, ICAM-2, VCAM-1, MMP-9 level in brain and serum samples were assessed. Histopathological examination and blood brain barrier permeability study was done.
Mean colitis score decreased in thalidomide treated group but not in ZNS treated group. Seizures were decreased in ZNS and Thali treated groups. Seizures were significantly higher in TNBS instilled group as compared to veh group. Oxidative stress was significantly lower in ZNS and Thali treated groups as compared to TNBS instilled group. TNF-α, IL-6, IL-1β, ICAM-1, VCAM-1, MMP-9 levels were found increased in TNBS instilled animals treated with vehicle whereas a reverse observation was noted for ICAM-2 level in brain and serum. Histopathological showed establishment of ulcerative colitis model in animals. Blood brain barrier was found distorted in TNBS instilled groups of animals. Seizure susceptibility may increase in experimental models of ulcerative colitis and anti-inflammatory agent may ameliorate this effect. doi:10.1016/j.vascn.2016.02.135
0137 Utility of an in vitro platform to assess neuronal toxicity using human iPSC-derived neurons Dinah Misnera, Chelsea Snyderc, Tin Ngob, Lanlan Yua, Nicola Stagga, Karin Staflina a
Genentech, South San Francisco, CA, USA University of California, Davis, Davis, CA, USA c Gilead Sciences, Foster City, CA, USA b
A major cause of drug attrition from clinical trials is neurotoxicity, a problem likely due to the lack of predictivity of currently available preclinical models to human safety. One of the most common druginduced neurotoxicities observed clinically is peripheral neuropathy. Existing in vivo animal models are time-consuming, expensive, and lack a direct species relationship to humans. Current in vitro models consist of immortalized cell lines and primary rodent cultures, which show variable correlation to human physiology. There is an unmet need for higher throughput, in vitro human-based assays that can identify compounds with neurotoxic and neuropathic potential. These in vitro assays can precede in vivo animal studies, and augment with data in the most relevant species before clinical trials begin. We chose to establish such assays with human induced pluripotent stem cells (iPSCs) that have been differentiated into mature neurons. These iPSC neurons offer a renewable source of mature cells that may be used to investigate neurotoxic potential. Our characterization of these neurons shows that they express mature markers at both transcript and protein levels. Using kinetic, high-throughput imaging we evaluated a large panel of test compounds known to cause peripheral neuropathy in the clinic and examined effects on neurite outgrowth and viability. Results from our assays show that these test compounds exert neurotoxicity in a classspecific manner and that our assessment correlates to clinical findings. We propose the use of this in vitro human assay to evaluate the potential for neurotoxicity and to provide utility in drug safety applications. doi:10.1016/j.vascn.2016.02.136
0138 NeuroSafe: A human integrated in vitro Neurotoxicity Safety Platform Loredana Redaellia, Giovanna Marescaa, Sara Tremoladaa, Christina Kuhna, Matteo Brioschia, Dietmar Hessb, Elke Guentherb, Lia Scarabottoloa