Bio-Techne Announces Travel Grants to SFN 2017

Temporal lobe epilepsy (TLE) is typified by multiple epileptogenic changes in the hippocampus, which include reductions in GABA-ergic interneurons, depletion of fibroblast growth factor-2 (FGF-2), waned neurogenesis, and chronic inflammation. These changes may contribute to cognitive, memory and mood dysfunction and spontaneous recurrent seizures (SRS). We examined the beneficial effects of combined grafting of medial ganglionic eminence derived neural stem cells (MGE-NSCs) into the hippocampus and two-weeks of systemic FGF-2 administration in a rat model of chronic TLE. We induced status epilepticus (SE) in young adult F344 rats via injections of kainic acid. Four months later, chronically epileptic rats (CERs) exhibiting comparable frequency of SRS were randomly assigned to one of 4 groups: CERs receiving MGE-NSC’s and FGF-2; CERs receiving MGE-NSC’s, CERs receiving sham surgery  and epilepsy-only controls. The first two CER groups received bilateral grafting of MGE-NSCs into the CA3 region of the hippocampus (3 grafts/side, 100,000 live cells/graft). Donor cells were isolated and expanded from MGEs dissected from gestation day 13 GFP-positive F344 rat fetuses. The first group also received daily SQ injections of FGF-2 (40ng/gm/day) for 2 weeks after grafting. Three to four months later (~8 months after SE), animals were examined for cognitive, memory and mood function using multiple behavioral tests followed by 3 weeks of continuous EEG recordings. Animals receiving MGE-NSCs plus FGF-2 or MGE-NSCs alone displayed improved ability for novel object recognition and pattern separation, and reduced depressive-like behavior, in comparison to CER control groups. Animals receiving MGE-NSCs plus FGF-2 also showed ability to discern minor changes in the environment in an object location test. Continuous EEG recordings uncovered 45-51% reduction in the frequency and intensity of SRS, and 65% reduction in the total time spent in seizure activity, in CERs receiving MGE-NSCs and FGF-2 or MGE-NSCs alone.  Graft-derived cells were seen in the CA3 region with a yield of 24-28% of injected cells, of which ~20% expressed GABA. Some graft-derived processes could also be seen in the adjoining CA1 subfield and the dentate gyrus. ~20% of graft-derived cells expressed GABA . Doublecortin immunostaining revealed increased neurogenesis in animals receiving MGE-NSCs and FGF-2. Thus, grafting of MGE-NSCs alone or MGE-NSCs with FGF-2 treatment can ease recognition memory, pattern separation and mood impairments and lessen SRS in CERs. However, FGF-2 treatment after grafting of MGE-NSCs also improves hippocampus-dependent location memory function and neurogenesis.

Acetylcholine (ACh) modulates the immune system activity and the inflammatory mediators by activating of muscarinic and nicotinic acetylcholine receptors expressed by immune cells. Cholinergic system dysfunction is present in MS patients and contributes to inflammatory processes. In fact, we demonstrated that in serum and CSF of MS patients, ACh levels were lower than in HD subjects. The ACh levels in MS patients are negatively correlated with the high levels of hydrolyzing enzymes AChE and BuChE. Moreover high levels of ACh hydrolyzing enzymes are positively correlated to the high levels of pro-inflammatory cytokines. Similarly, in brain of EAE mice we observed low levels of ACh biosynthetic enzyme ChAT couple with high levels of AChE and BuChE comparing mice in relapse vs remitting phase. These results indicate that the decreased levels of ACh may contribute to exacerbate the inflammatory state in MS. Neuroinflammation plays a critical role in several nervous system disorders. The microglia in the CNS and the immune cells in the blood, respond to ACh stimulation, modulating the inflammatory mediators by alph- 7 nicotinic receptors (nAChRs) activation. It has been reported that nicotine treatment significantly delays and attenuates inflammatory and autoimmune responses in EAE. Moreover we recently demonstrated that the stimulation of nAChRs by nicotine was able to decrease the IL-1beta and IL-17 expression in PBMC of MS patients. These and other results highlight the role of the alpha-7 receptor subtype in the modulation of inflammatory cytokines. These receptors represent an innovative approach in the treatment of those neuropathologies in which local inflammation plays a crucial role, including MS. Recent significant drug discovery efforts have been devoted to nAChRs and several promising selective ligands have been developed. In this respect, the identification of potent selective alpha-7 nAChRs ligands showing minimal or no side effects and able to cross the blood brain barrier, is of strategic relevance for the therapeutic treatment of several nervous system disorders. In the present project we propose to investigate the ability of novel alpha7 nAChRs agonists ((R)-(-)-ICH-3 and (R)-(-)-ICH-4) to modulate the cholinergic system activity and peripheral inflammation. Moreover, by the collaboration with a group of researchers in Edinburg, using the induced pluripotent stem cells (iPS) obtained from human fibroblasts, we can produce oligodendrocytes from healthy subjects and MS patients. This approach may allow to study for the first time, the effects produced by cholinergic stimulation on human oligodendrocytes in terms of proliferation, survival and differentiation.

The data obtained might contribute to better characterize the role of a7 nAChRs and cholinergic stimulation in the modulation of inflammatory processes and oligodendrocytes survival and differentiation, and to identify and propose new therapeutic tools for MS treatment 

Increased HIV RNA replication and pro-inflammatory cytokines and proteins are hallmarks of untreated HIV infection progression. HIV infected persons may present with subtle neurocognitive impairment with little or no functional impairment in activities of daily living (ADL) during the early to middle stages of the disease; and this tends to affect their quality of life. Neurocognitive impairment results from HIV-induced proinflammatory neural toxicity due a chronic persistent micronutrient deficient state. Pro-inflammatory cytokines induce increased demand for and consumption of anti-oxidants and other micronutrients thus maintaining a micronutrient deficient state among HIV-infected persons irrespective of their clinical stage.    Therefore it is postulated that micronutrients supplementation may retard HIV induced hyper-catabolism; improve oxidative and nutritional status of patients, delay progression of HIV infection to AIDS, and improve mental function of infected persons. This research studied the effect of micronutrient supplementation on HIV RNA load, CD4+ T cell count, IL-6 and C-reactive proteins in untreated HIV infected patients.

We recruited 90 antiretroviral naïve adult patients at our hospital HIV clinics after informed consent. Inclusion criteria were: stage 1 and 2, CD4+ >500 cells/µl, absence of immunosuppressive and/or psychiatric illness. Their mental status were evaluated using the Mental Status Questionnaire (MSQ), Mini-mental State examination (MMSE), modified HIV Dementia scale (mHDS), after which plasma HIV RNA load, blood CD4+ counts, serum interleukin-6 (IL-6) and C-reactive proteins (CRP) were determined at baseline. Each patient was given one capsule of SYNOVITTM (micronutrient supplements) to take daily; and at 24 weeks of SYNOVITTM supplementation, the patients were re-evaluated. Patients who became pregnant, defaulted from one clinic visit or whose CD4+ reduced below 500 cells/µl were excluded.

At 24 weeks of SYNOVITTM supplementation, MSQ, MMSE and MDS scores increased respectively from 8.0 to 10.0 (p>0.05), 19.0 ± 0.2 to 27.2 ± 0.6 (p<0.05) and 6.0 ±1.2 to 11.0 ± 1.2 (p<0.05); however, HIV RNA load (copies/ml) reduced from 27,105.0 to 16,227.5 (p>0.05); IL-6 (pg/ml) from to 1.3 (p>0.05); CRP (ng/ml) 29.4 to 26.2 (p<0.05); and CD4+ (cells/µl) from 704.5 to 561.0 (p<0.05). In conclusion, micronutrient supplementation improved mental status and reduced HIV RNA replication, pro-inflammatory biomarkers (IL-6, CRP) of HIV infection but did not improve CD4+ cell counts.

Lupus is a disease for which effective therapy is an urgent need. The current medications such as cyclophophamide have toxic side effects. Complement activation is an important aspect of the disease and forma a part of the clinical profile. Recent studies reveal novel roles for the different complement proteins along with the traditionally understood immune functions. One of the first organs affected in lupus patients is the brain. Using current technology and both, the mouse in vivo and the cultured cells in vitro we are studying the role of complement proteins in this devastating disease. To increase the awareness that complement proteins are potential therapeutic targets, to exchange ideas about their mechanisms of action and their links with other signaling pathways, the travel grant would be a godsend.