Data-driven learning how oncogenic gene expression locally alters heterocellular networks

Developing drugs increasingly relies on mechanistic modeling and simulation. Models that capture causal relations among genetic drivers of oncogenesis, functional plasticity, and host immunity complement wet experiments. Unfortunately, formulating such mechanistic cell-level models currently relies on hand curation, which can bias how data is interpreted or the priority of drug targets. In modeling molecular-level networks, rules and algorithms are employed to limit a priori biases in formulating mechanistic models. Here we combine digital cytometry with Bayesian network inference to generate causal models of cell-level networks linking an increase in gene expression associated with oncogenesis with alterations in stromal and immune cell subsets from bulk transcriptomic datasets. We predict how increased Cell Communication Network factor 4, a secreted matricellular protein, alters the tumor microenvironment using data from patients diagnosed with breast cancer and melanoma. Predictions are then tested using two immunocompetent mouse models for melanoma, which provide consistent experimental results

Immunogenicity and antitumor activity of the superlytic λF7 phage nanoparticles displaying a HER2/neu-derived peptide AE37 in a tumor model of BALB/c mice

The final publication is available at Elsevier via http://dx.doi.org/10.1016/j.canlet.2018.03.030 © 2018. This manuscript version is made available under the CC-BY-NC-ND 4.0 license https://creativecommons.org/licenses/by-nc-nd/4.0/Phage display technique has been increasingly researched for vaccine design and delivery strategies in recent years. In this study, the AE37 (Ii-Key/HER-2/neu 776–790) peptide derived from HER2 (human epidermal growth factor receptor protein) was used as a fused peptide to the lambda phage (λF7) coat protein gpD, and the phage nanoparticles were used to induce antitumor immunogenicity in a TUBO model of breast cancer in mice. Mice were immunized with the AE37 peptide displaying phage, λF7 (gpD::AE37) every 2-week intervals over 6-weeks, then the generated immune responses were evaluated. An induction of CTL immune response by the λF7 (gpD::AE37) construct compared to the control λF7 and buffer groups was observed in vitro. Moreover, in the in vivo studies, the vaccine candidate showed promising prophylactic and therapeutic effects against the HER2 overexpressing cancer in BALB/c mice.Mashhad University of Medical Sciences, Mashhad, Iran bach (MUMS GN: 922610)NSERC, Canada (NSERC GN: 214684

Preparation of nanoliposomes containing HER2/neu (P5+435) peptide and evaluation of their immune responses and anti-tumoral effects as a prophylactic vaccine against breast cancer.

HER2/neu is an immunogenic protein inducing both humoral and cell-mediated immune responses. The antigen-specific cytotoxic T lymphocytes (CTLs) are the main effector immune cells in the anti-tumor immunity. To induce an effective CTL specific response against P5+435 single peptide derived from rat HER2/neu oncogene, we used a liposome delivery vehicle. In vivo enhancement of liposome stability and intracytoplasmic delivery of peptides are the main strategies which elevate the liposome-mediated drug delivery. Liposomes containing high transition temperature phospholipids, such as DSPC, are stable with prolonged in vivo circulation and more accessibility to the immune system. Incorporation of DOPE phospholipid results in the effective delivery of peptide into the cytoplasm via the endocytotic pathway. To this end, the P5+435 peptide was linked to Maleimide-PEG2000-DSPE and coupled on the surface of nanoliposomes containing DSPC: DSPG: Cholesterol with/without DOPE. We observed that mice vaccinated with Lip-DOPE-P5+435 formulation had the highest number of IFN-γ- producing CTLs with the highest cytotoxic activity that consequently led to significantly smallest tumor size and prolonged survival rate in the TUBO mice model. In conclusion, our study indicated that the liposomal form of P5+435 peptide containing DOPE can be regarded as a promising prophylactic anti-cancer vaccine to generate potent antigen-specific immunity

Conjugated nanoliposome with the HER2/neu-derived peptide GP2 as an effective vaccine against breast cancer in mice xenograft model - Fig 2

<p><b>Splenocyte cell phenotype and level of cytokine expression (A and B) and lymph nodes (C) of BALB / c mice immunized with different liposomal formulations.</b> 14 days after the last immunization, splenocytes were isolated and stimulated <i>in vitro</i> with PMA/I for 4h and stained with a surface CD8 and CD4 marker and intracellular IFN-γ and IL-4 cytokine prior to FACS analysis. (A)Geometric mean fluorescence intensity (MFI) level for INF-γ in gated CD8 and CD4 in the spleen. (B)MFI level for IL-4 in gated CD4 lymphocyte populations in the spleen. (C) MFI level for INF-γ in gated CD8, CD4, and IL-4 in gated CD4 in lymph nodes. Data represent mean± SEM (= 3).*<i>p</i><0.05, **<i>p</i><0.01, and ***<i>p</i><0.001; denote significant difference from all other formulations and the HEPS-dextrose 5%.</p

Prophylactic efficacy data of different liposomal vaccine formulations in TUBO tumor model of mice (n = 5).

<p>Prophylactic efficacy data of different liposomal vaccine formulations in TUBO tumor model of mice (n = 5).</p

qRT-RCR.

<p>Analysis of IFN-γ and IL-4 levels in splenocytes isolated from BALB/c mice vaccinated with Lip-DOPE-MPL-GP2, two weeks after the final vaccination. All values represent means ± SD (n = 3).</p

Therapeutic efficacy data for liposomal formulations in TUBO tumor mice model (n = 5).

<p>Therapeutic efficacy data for liposomal formulations in TUBO tumor mice model (n = 5).</p

Characteristics of liposomal formulations (n = 3; Mean ±SD).

<p>Characteristics of liposomal formulations (n = 3; Mean ±SD).</p