2017 Targeting Trends Review

Hay C, Sult E, Huang Q, Mulgrew K, Fuhrmann S, McGlinchey K, Hammond S, Rothstein R, Rios-Doria J, Poon E, Holoweckyj N, Durham N, Leow C, Diedrich G, Damschroder M, Herbst R, Hollingsworth R, Sachsenmeier K (2016) Targeting CD73 in the tumor microenvironment with MEDI9447. Oncoimmunology 5:e1208875. doi: 10.1080/2162402X.2016.1208875

Summary: MEDI9447 is a human monoclonal antibody that is specific for the ectoenzyme CD73 and currently undergoing Phase I clinical trials. Here the authors show that MEDI9447 is a potent inhibitor of CD73 ectonucleotidase activity, with wide ranging immune regulatory consequences. MEDI9447 results in relief from adenosine monophosphate (AMP)-mediated lymphocyte suppression in vitro and inhibition of mouse syngeneic tumor growth in vivo. In contrast with other cancer immunotherapy agents such as checkpoint inhibitors or T-cell agonists, MEDI9447 drives changes in both myeloid and lymphoid infiltrating leukocyte populations within the tumor microenvironment of mouse models. In vitro experiments validating the internalization of antibodies into cell lines MDA-MB-231 and 4T1 were measured using the Fab-ZAP human antibody internalization kit (Cat. #KIT-51-Z). Combination data showing additive activity between MEDI9447 and anti-PD-1 antibodies using human cells in vitro and mouse tumor models further demonstrate the potential value of relieving adenosine-mediated immunosuppression. Based on these data, a Phase I study to test the safety, tolerability, and clinical activity of MEDI9447 in cancer patients was initiated (NCT02503774).

Related Products: Fab-ZAP human (Cat. #IT-51)

Jiao J, Xiong W, Wang L, Yang J, Qiu P, Hirai H, Shao L, Milewicz D, Chen Y, Yang B (2016) Differentiation defect in neural crest-derived smooth muscle cells in patients with aortopathy associated with bicuspid aortic valves. EBioMedicine 10:282-290. doi: 10.1016/j.ebiom.2016.06.045 PMID: 27394642

Summary: Individuals with bicuspid aortic valves (BAV) are at a higher risk of developing thoracic aortic aneurysms (TAA) than patients with trileaflet aortic valves (TAV). Aneurysms associated with BAV most commonly involve the ascending aorta. Smooth muscle cells (SMCs) in the ascending and descending aorta arise from neural crest (NC) and paraxial mesoderm (PM), respectively. Scientists hypothesized defective differentiation of the neural crest stem cells (NCSCs)-derived SMCs but not paraxial mesoderm cells (PMCs)- derived SMCs contributes to the aortopathy associated with BAV. Induced pluripotent stem cells (iPSCs) from BAV/TAA patients were differentiated into NCSC-derived SMCs and showed decreased expression of a marker of SMC differentiation (MYH11) and impaired contraction. The scientists demonstrated that decreased differentiation and contraction of patient’s NCSC-derived SMCs may contribute to the aortopathy associated with BAV.

Usage: Anti-NGFr (ME20.4, p75, Cat. #AB-N07) was used for the immunofluorescence staining and flow cytometry of NCSCs.

Related Products: NGFr (ME20.4, p75) Mouse Monoclonal (Cat. #AB-N07)

Wehner A, Milen A, Albin R, Pierchala B (2016) The p75 neurotrophin receptor augments survival signaling in the striatum of pre-symptomatic Q175(WT/HD) mice. Neuroscience 324:297-306. doi: 10.1016/j.neuroscience.2016.02.069 PMID: 26947127

Summary: Huntington’s disease (HD) is a dominantly inherited neurodegenerative disorder. It’s characterized by a combination of motor, cognitive, and psychiatric features. Striatal spiny neurons are dependent on brain-derived neurotropic factor for proper function and survival. Studies suggest both the receptors for BDNF, TrkB and the p75 neurotrophin receptor (p75), are improperly regulated in the striata of HD patients and mouse models. The authors investigated the role of p75 in the Q175 knock-in mouse model of HD be examining levels of activation of downstream signaling molecules to determine if p75 represents a promising therapeutic target. Anti-NGFr (mup75) (Cat. #AB-N01AP) was used at a 1:2000 dilution in immunoblotting. The data suggest that p75 signaling plays an early role in augmenting pro-survival signaling in the striatum and that disruption of p75 signaling at a pre-symptomatic age may exacerbate pathologic changes in these knock-in mouse models.

Related Products: NGFr (mu p75) Rabbit Polyclonal, affinity-purified (Cat. #AB-N01AP)

Iizuka K, Yokomizo T, Watanabe N, Tanaka Y, Osato M, Takaku T, Komatsu N (2016) Lack of phenotypical and morphological evidences of endothelial to hematopoietic transition in the murine embryonic head during hematopoietic stem cell emergence. PLoS One 11:e0156427. doi: 10.1371/journal.pone.0156427 PMID: 27227884

Summary: Hemogenic endothelial cells have been observed in several embryonic tissues, such as the dorsal aorta, the placenta and the yolk sac. Recent work also suggest that the mouse embryonic head also produces hematopoietic stem cells (HSCs)/progenitors. However, a histological basis for HSC generation in the head hasn’t been determined because the hematopoietic cluster and hemogenic endothelium have not been well characterized. The authors in this study used whole-mount immunostaining and 3D confocal reconstruction techniques to analyze both c-Kit hematopoietic cluster and Runx1 hemogenic endothelium in the whole-head vasculature. Alexa488 labeled anti-NGFr (Cat. #FL-N01AP) was used in flow cytometry. The number of c-Kit hematopoietic cells was 20-fold less in the head arteries than in the dorsal aorta. In addition, nascent hematopoietic cells, observed by a budding structure and a Runx1 hemogenic endothelium, were not observed in the head. These results suggest that head HSCs may not be or are rarely generated from the endothelium in the same manner as aortic HSCs.

Related Products: NGFr (mu p75) Rabbit Polyclonal, affinity-purified Alexa488-labeled (Cat. #AB-N01APFLA)

Wang L, Conner J, Nagahara A, Tuszynski M (2016) Rehabilitation drives enhancement of neuronal structure in functionally relevant neuronal subsets. Proc Natl Acad Sci U S A 113:2750-2755. doi: 10.1073/pnas.1514682113

Summary: Rehabilitation is often prescribed after brain injury, but the basis for how training can influence brain plasticity and recovery is unclear. In this study, the authors show that intense rehabilitation training after focal brain injury drives significant structural changes in brain cells located adjacent to the injury. Importantly, a key brain modulatory system, the basal forebrain cholinergic system, is required for enabling rehabilitation to impact brain structure. Rats underwent cholinergic ablations by injecting 192-IgG-Saporin (Cat. #IT-01) into the nucleus basalis (0.2-0.25 mcl of 0.375 mg/ml solution in artificial CSF). Damage to the cholinergic system, which can occur naturally during aging, completely blocks brain plasticity mediated by rehabilitation and significantly attenuates functional recovery. These results provide new insights into how rehabilitation may promote recovery and suggest that brain cholinergic systems may be a possible therapeutic target for influencing recovery.

Related Products: 192-IgG-SAP (Cat. #IT-01)

Tanigawa M, Suzuki C, Niwano K, Kanekatsu R, Tanaka H, Horiike K, Hamase K, Nagata Y (2016) Participation of D-serine in the development and reproduction of the silkworm Bombyx mori. J Insect Physiol 87:20-29. doi: 10.1016/j.jinsphys.2016.01.006 PMID: 26828952

Summary: The silkworm Bombyx mori is known to contain high levels of free D-serine, an optical isomer of L-serine. In this work, the authors investigated the localization of D-serine in various organs of the silkworms in various stages of life in an effort to elucidate its role. They used an immunohistochemical approach to localize D-serine to the silkworm hemolymph, midgut, testes, ovaries, and fat bodies. They also used rabbit antibody against glutaraldehyde-conjugated D-alanine (Cat. #AB-T049) to examine the distribution of D-alanine throughout the silkworms. The authors treated silkworms with an inhibitor of serine racemase to reduce the conversion of L- to D-serine and on the basis of their results suggested the possible involvement of D-serine in ATP synthesis for metamorphosis and reproduction.

Sakurai T, Kamiyoshi A, Kawate H, Mori C, Watanabe S, Tanaka M, Uetake R, Sato M, Shindo T (2016) A non-inheritable maternal Cas9-based multiple-gene editing system in mice. Sci Rep 6:20011. doi: 10.1038/srep20011

Summary: In this work, the authors generated transgenic mice with systemic Cas9 overexpression (Cas9 mice) in order to simplify the procedure of generating genetically modified animals using the CRISPR/Cas9 system – only guide RNAs (gRNAs) would need to be administered to induce mutations at target loci. To test Cas9 mice for genome editing in vitro, the authors transiently transfected primary fibroblasts from Cas9 mice with Ggta1 gRNA (Ggta1 is responsible for synthesizing the cell-surface α-Gal epitope). They treated the fibroblasts with rIB4-SAP (Cat. #IT-10) and found that it killed Ggta1 +/+ and KO/+ cells, while biallelic Ggta1 KO cells survived as they did not synthesize the α-Gal epitope. This indicated that primary cells from the Cas9 transgenic mice have CRISPR/Cas9 genome editing capability with the administration of gRNA alone. The success of their experiments indicate that this method could potentially be used to generate other genetically modified animals.

Related Products: IB4-SAP (Cat. #IT-10)

Xia W, Liu Y (2016) Targeted ablation of cardiac sympathetic neurons: A promising approach to prevent sudden cardiac death. Int J Cardiol 202:425-426. doi: 10.1016/j.ijcard.2015.09.049

Summary: Sudden cardiac death (SCD) refers to an unexpected death due to cardiovascular causes, occurring generally within 1 hr of symptom onset, in a person without any prior condition that would appear fatal. Currently, the implantable cardioverter-defibrillator (ICD) has been shown to be the most effective therapy for preventing SCD. However, the occurrence of lead complications is significant and more importantly, the ICD implantation remains costly and the quality of life for recipients is significantly affected with appropriate and inappropriate shocks. Sympathetic activation plays an important role in the pathophysiology of arrhythmias leading to SCD, and neuraxial modulation is emerging as an important avenue of therapeutic intervention. The authors demonstrate that targeted ablation of cardiac sympathetic neurons by bilateral stellate ganglia injection of CTB-SAP (Cat. #IT-14) is a novel method for sympathetic blockade. CTB-SAP will be retrogradely transported to the plasma membranse of sympathetic preganglionic neurons (SPNs) and bind to the GM1 gangliosides and subsequently ablate these neurons. Targeted ablation of cardiac sympathetic neurons by injection of CTB-SAP avoids the limitations of medical therapy and thoracic sympathectomy, such as incomplete compliance, Horner’s syndrome, and compensatory hyperhidrosis. Furthermore, they found that targeted ablation of cardiac sympathetic neurons reduces resting, reflex and exercise-induced sustained ventricular tachycardia, associated with a reduced number of neurons in the stellate ganglia and spinal cord, as well as a reduced left ventricular norepinephrine content and sympathetic innervation density. Therefore, targeted ablation of cardiac sympathetic neurons may be a promising approach to prevent SCD via regulating the cardiac autonomic nervous system.

Related Products: CTB-SAP (Cat. #IT-14)

Bourane S, Duan B, Koch S, Dalet A, Britz O, Garcia-Campmany L, Kim E, Cheng L, Ghosh A, Ma Q, Goulding M (2015) Gate control of mechanical itch by a subpopulation of spinal cord interneurons. Science 350:550-554. doi: 10.1126/science.aac8653

Summary: Light mechanical stimulation of the hairy skin can induce a form of itch known as mechanical itch. This itch sensation is normally suppressed by inputs from mechanoreceptors, however, in many forms of chronic itch, including alloknesis, this gating mechanism is lost. Scientists demonstrated that a population of spinal inhibitory interneurons (INs), that are defined by the expression of neuropeptide Y::Cre (NPY::Cre), act to gate mechanical itch. Mice in which dorsal NPY::Cre-derived neurons are selectively ablated or silenced develop mechanical itch without an increase in sensitivity to chemical itch or pain. This chronic itch state is histamine-independent and is transmitted independently of the GRP-GRPR signaling pathway. The scientists thereby revealed a dedicated spinal cord inhibitory pathway that gates the transmission of mechanical itch. Mice were given an intrathecal injection of 400 ng of Bombesin-SAP (Cat. #IT-40) in 10 ml of sterile saline to ablate GRPR-expressing neurons.

Related Products: Bombesin-SAP (Cat. #IT-40)

Sorge R, Mapplebeck J, Rosen S, Beggs S, Taves S, Alexander J, Martin L, Austin J, Sotocinal S, Chen D, Yang M, Shi X, Huang H, Pillon N, Bilan P, Tu Y, Klip A, Ji R, Zhang J, Salter M, Mogil J (2015) Different immune cells mediate mechanical pain hypersensitivity in male and female mice. Nat Neurosci 18:1081-1083. doi: 10.1038/nn.4053

Summary: A large and rapidly increasing body of evidence indicates that microglia-to-neuron signaling is essential for chronic pain hypersensitivity. Using multiple approaches, the authors found that microglia are not required for mechanical pain hypersensitivity in female mice; female mice achieved similar levels of pain hypersensitivity using adaptive immune cells, likely T lymphocytes. This sexual dimorphism suggests that male mice cannot be used as proxies for females in pain research. Mac-1-SAP mouse/human toxin (Cat. #IT-06, 15 μg in 8.8 μl) and Saporin control (Cat. #PR-01, 8.8 μg in 8.8 μl) were administered via i.t. injection. The topic of immune system involvement in chronic pain pathophysiology is one of the most active in the pain field; that this sex difference has not been observed until now is very surprising indeed. An important implication of the current findings is that distinct strategies targeting neuroimmune signaling might be required for the treatment of chronic pain in men versus women.

Related Products: Mac-1-SAP mouse/human (Cat. #IT-06), Saporin (Cat. #PR-01)