Gene Regulation Using Spherical Nucleic Acids to Treat Skin Disorders

Spherical nucleic acids (SNAs) are nanostructures consisting of nucleic acids in a spherical configuration, often around a nanoparticle core. SNAs are advantageous as gene-regulating agents compared to conventional gene therapy owing to their low toxicity, enhanced stability, uptake by virtually any cell, and ability to penetrate the epidermal barrier. In this review we: (i) describe the production, structure and properties of SNAs; (ii) detail the mechanism of SNA uptake in keratinocytes, regulated by scavenger receptors; and (iii) report how SNAs have been topically applied and intralesionally injected for skin disorders. Specialized SNAs called nanoflares can be topically applied for gene-based diagnosis (scar vs. normal tissue). Topical SNAs directed against TNFα and interleukin-17A receptor reversed psoriasis-like disease in mouse models and have been tested in Phase 1 human trials. Furthermore, SNAs targeting ganglioside GM3 synthase accelerate wound healing in diabetic mouse models. Most recently, SNAs targeting toll-like receptor 9 are being used in Phase 2 human trials via intratumoral injection to induce immune responses in Merkel cell and cutaneous squamous cell carcinoma. Overall, SNAs are a valuable tool in bench-top and clinical research, and their advantageous properties, including penetration into the epidermis after topical delivery, provide new opportunities for targeted therapies.

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siRNA-based spherical nucleic acids reverse impaired wound healing in diabetic mice by ganglioside GM3 synthase knockdown

Proceedings of the National Academy of Sciences ◽

10.1073/pnas.1505951112 ◽

2015 ◽

Vol 112 (18) ◽

pp. 5573-5578 ◽

Cited By ~ 100

Author(s):

Pratik S. Randeria ◽

Mark A. Seeger ◽

Xiao-Qi Wang ◽

Heather Wilson ◽

Desmond Shipp ◽

...

Keyword(s):

Wound Healing ◽

Nucleic Acids ◽

Egf Receptor ◽

Receptor Activation ◽

Diabetic Mice ◽

Ganglioside Gm3 ◽

Intact Mouse ◽

Impaired Wound Healing ◽

Igf1 Receptor ◽

Spherical Nucleic Acids

Spherical nucleic acid (SNA) gold nanoparticle conjugates (13-nm-diameter gold cores functionalized with densely packed and highly oriented nucleic acids) dispersed in Aquaphor have been shown to penetrate the epidermal barrier of both intact mouse and human skin, enter keratinocytes, and efficiently down-regulate gene targets. ganglioside-monosialic acid 3 synthase (GM3S) is a known target that is overexpressed in diabetic mice and responsible for causing insulin resistance and impeding wound healing. GM3S SNAs increase keratinocyte migration and proliferation as well as insulin and insulin-like growth factor-1 (IGF1) receptor activation under both normo- and hyperglycemic conditions. The topical application of GM3S SNAs (50 nM) to splinted 6-mm-diameter full-thickness wounds in diet-induced obese diabetic mice decreases local GM3S expression by >80% at the wound edge through an siRNA pathway and fully heals wounds clinically and histologically within 12 d, whereas control-treated wounds are only 50% closed. Granulation tissue area, vascularity, and IGF1 and EGF receptor phosphorylation are increased in GM3S SNA-treated wounds. These data capitalize on the unique ability of SNAs to naturally penetrate the skin and enter keratinocytes without the need for transfection agents. Moreover, the data further validate GM3 as a mediator of the delayed wound healing in type 2 diabetes and support regional GM3 depletion as a promising therapeutic direction.

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Pengaruh Gel Ekstrak Daun Kerehau (Callicarpa longifolia Lam.) Terhadap Penyembuhan Luka Pada Model Tikus Diabetes

Kartika Jurnal Ilmiah Farmasi ◽

10.26874/kjif.v6i2.154 ◽

2019 ◽

Vol 6 (2) ◽

pp. 70

Author(s):

Sansan Candra ◽

Elis Susilawati ◽

I Ketut Adnyana

Keyword(s):

Wound Healing ◽

Mouse Models ◽

Leaf Extract ◽

Healing Process ◽

Diabetic Mouse ◽

Diabetic Animal ◽

Wound Healing Process ◽

Diabetic Ulcer ◽

Average Percentage ◽

Accelerate Wound Healing

Abstrak Luka diabetes adalah infeksi, luka dan destruksi pada penderita diabetes melitus. Daun kerehau secara empiris digunakan oleh suku Dayak Tunjung sebagai obat luka dan bengkak. Penelitian ini bertujuan untuk mengetahui pengaruh gel ekstrak daun kerehau terhadap penyembuhan luka pada model tikus diabetes. Induksi diabetes dilakukan menggunakan aloksan dosis 75 mg/kg BB intravena. Model hewan diabetes dibuat luka dengan kedalaman 0,3 cm menggunakan scalpel steril pada bagian punggung. Hewan uji dibagi menjadi lima kelompok dan mendapatkan sediaan basis gel, Prontosan® gel, sediaan gel ekstrak daun kerehau konsentrasi 2,5%, 5%, dan 10%. Pengolesan gel dilakukan dua kali sehari selama 13 hari. Parameter penyembuhan luka dilakukan dengan mengukur panjang luka dan dinyatakan dengan persentase penyembuhan luka. Hasil pengujian menunjukkan bahwa dengan pemberian gel ekstrak daun kerehau dapat mempercepat penyembuhan luka pada model tikus diabetes dimana sediaan gel ekstrak daun kerehau konsentrasi 2,5% memberikan kesembuhan pada hari ke-13, sediaan gel konsentrasi 5% pada hari ke-12, dan sediaan gel konsentrasi 10% pada hari ke-9. Hasil juga menunjukkan rata-rata persentase penyembuhan luka pada kelompok konsentrasi 10% lebih tinggi dibandingkan konsentrasi 2,5% dan 5%. Dapat disimpulkan bahwa pemberian gel ekstrak daun kerehau dapat menyembuhkan luka pada model tikus diabetes. Kata kunci: Luka diabetes, daun kerehau, penyembuhan luka Effect of Kerehau Leaves Extract Gel to Wound Healing Process in Diabet Rats Model Abstract Diabetic wounds are infection, injury and destruction in people with diabetes mellitus. Kauhau leaves are empirically used by the Tunjung Dayak tribe as a medicine for wounds and swelling. The aim of this study was to determine the effect of Kerehau leaf extract gel on wound healing in diabetic mouse models. Induction of diabetes is done using alloxan dose of 75 mg / kg BW intravenously. The diabetic animal model was made with a 0.3 cm depth wound using a sterile scalpel on the back. The test animals were divided into five groups and obtained base gel preparations, Prontosan® gel, preparations of Kerehau leaf extract gel concentrations of 2.5%, 5%, and 10%. Gel application is done twice a day for 13 days. Wound healing parameters were performed by measuring the length of the wound and expressed by the percentage of wound healing. The test results showed that administration of Kerehau leaf extract gel could accelerate wound healing in diabetic mouse models where the preparations of Kerehau leaf extract gel with a concentration of 2.5% gave healing on the 13th day, preparations for 5% gel on 12th day, and preparations gel concentration of 10% on day 9. The results also showed that the average percentage of wound healing in the concentration group was 10% higher than the concentration of 2.5% and 5%. It can be concluded that administration of Kerehau leaf extract gel can heal wounds in diabetic mouse models. Keywords: Diabetic ulcer, kerehau leaf, wound healing

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627 The DLL3-targeted half-life extended bispecific T cell engager (HLE BiTE®) immune-oncology therapy AMG 757 has potent antitumor activity in neuroendocrine cancer

Journal for ImmunoTherapy of Cancer ◽

10.1136/jitc-2020-sitc2020.0627 ◽

2020 ◽

Vol 8 (Suppl 3) ◽

pp. A663-A663

Author(s):

Keegan Cooke ◽

Juan Estrada ◽

Jinghui Zhan ◽

Jonathan Werner ◽

Fei Lee ◽

...

Keyword(s):

T Cells ◽

T Cell ◽

Tumor Growth ◽

Mouse Models ◽

T Cell Activation ◽

Phase 1 ◽

Phase 1 Study ◽

Human T Cells

BackgroundNeuroendocrine tumors (NET), including small cell lung cancer (SCLC), have poor prognosis and limited therapeutic options. AMG 757 is an HLE BiTE® immune therapy designed to redirect T cell cytotoxicity to NET cells by binding to Delta-like ligand 3 (DLL3) expressed on the tumor cell surface and CD3 on T cells.MethodsWe evaluated activity of AMG 757 in NET cells in vitro and in mouse models of neuroendocrine cancer in vivo. In vitro, co-cultures of NET cells and human T cells were treated with AMG 757 in a concentration range and T cell activation, cytokine production, and tumor cell killing were assessed. In vivo, AMG 757 antitumor efficacy was evaluated in xenograft NET and in orthotopic models designed to mimic primary and metastatic SCLC lesions. NSG mice bearing established NET were administered human T cells and then treated once weekly with AMG 757 or control HLE BiTE molecule; tumor growth inhibition was assessed. Pharmacodynamic effects of AMG 757 in tumors were also evaluated in SCLC models following a single administration of human T cells and AMG 757 or control HLE BiTE molecule.ResultsAMG 757 induced T cell activation, cytokine production, and potent T cell redirected killing of DLL3-expressing SCLC, neuroendocrine prostate cancer, and other DLL3-expressing NET cell lines in vitro. AMG 757-mediated redirected lysis was specific for DLL3-expressing cells. In patient-derived xenograft and orthotopic models of SCLC, single-dose AMG 757 effectively engaged human T cells administered systemically, leading to a significant increase in the number of human CD4+ and CD8+ T cells in primary and metastatic tumor lesions. Weekly administration of AMG 757 induced significant tumor growth inhibition of SCLC (figure 1) and other NET, including complete regression of established tumors and clearance of metastatic lesions. These findings warranted evaluation of AMG 757 (NCT03319940); the phase 1 study includes dose exploration (monotherapy and in combination with pembrolizumab) and dose expansion (monotherapy) in patients with SCLC (figure 2). A study of AMG 757 in patients with neuroendocrine prostate cancer is under development based on emerging data from the ongoing phase 1 study.Abstract 627 Figure 1AMG 757 Significantly reduced tumor growth in orthotopic SCLC mouse modelsAbstract 627 Figure 2AMG 757 Phase 1 study designConclusionsAMG 757 engages and activates T cells to kill DLL3-expressing SCLC and other NET cells in vitro and induces significant antitumor activity against established xenograft tumors in mouse models. These preclinical data support evaluation of AMG 757 in clinical studies of patients with NET.Ethics ApprovalAll in vivo work was conducted under IACUC-approved protocol #2009-00046.

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