Tools and Avenues for Nanotechnology-based Vectors Exploitation for Biomarker Signature and Therapeutical Drug Delivery

Nanomedicine main goal is to ameliorate biodistribution and side effects of therapeutics. Proper Micro (MP) and Nanoparticles (NP) can i) protect therapeutic molecules from reticuloendothelial clearance, ii) transport them to the site of action minimizing their influence on normal tissues and iii) enhance drug concentration and effects in target tissues and cells, allowing the use of lower doses. However a single component nanovector cannot accomplish all those duties; therefore, our efforts are focused on the optimization and performance evaluation of multi-component drug-delivering nanosystems, made of silica, silicon, polymers, lipids and proteins from synthetic and biological sources. In recent years, the groups in Naples and in Houston in cooperation have studied many different strategies which permit MP and NP to overcome one or more biological barriers (BB) that hamper the delivery of therapeutics to the desired site of action. The two research groups have produced a series of results which will be briefly summarized in this report and then discussed in greater detail in the talk. 1) Diverse types of Multistage vector (MSV) were produced and used to enhance the delivery of free and encapsulated drugs in cellular and mice models of cancer. MSV is constituted by a silicon mesoporous microparticle which protect nanoparticles or free drugs from reticuloendothelial clearance, superbly marginates toward vessel walls and preferentially accumulates on inflamed endothelium [1-3]. 2) A Biomimetic drug delivery platform was produced with a nanoporous silicon core and and a shell derived from the leukocyte cell membrane. This Leukolike vector (LLV) and it’s therapeutic potentialities were extensively investigated in vitro and in vivo, from a biological and biochemical point of view. LLV shows the ability to: 1) evade the immune system; 2) circulate longer in the blood stream; 3) communicate with endothelial cells through receptor–ligand interactions increasing endothelium permeability; 4) transport and release a payload across inflamed endothelium; 5) accumulate in a tumour [4-7]. 3) Innovative microfluidic systems were developed to mimick blood capillary circulation, in order to study and to predict the in flow dynamics and the margination tendency of different types of MP, also in presence of blood cells, using real human blood as circulating solution [8]. 4) Enzyme-functionalized silica NPs were conceived and produced to digest tumor extracellular matrix, in vitro and in vivo achieving a better penetration in the tumoral tissue [9]. 5) pH-responsive hybrid nanoparticles (HNP) were conceived, produced, characterized and successfully used to achieve efficient siRNA delivery in cell culture and in mice models of human breast cancer. HNP are constituted by a shell of cationic hydrogel able to electrostatically bind siRNA and by a supporting nanostructured core of silica that provides mechanical stability to the system. HNP are able to escape from endolysosomal compartment through a proton sponge effect [5,10]. In the light of what has been briefly outlined, the following avenues will be pursued in the near future and will be discussed: (i) Production and evaluation of different types of MSV for cancer therapy (ii) Protein and peptide functionalization of NPs to enhance tumor recognition and penetration (iii) Production and evaluation of different types of NP for siRNA delivery (iv) Production of Organ-on-chip microfluidic devices able to study and predict the fluidic and biological performance of MP and NP. (v) Characterization of plasma Protein Corona (PC) [11] of MP and NP by proteomic approaches and correlations between PC composition and targeting of specific tissues. References [1] Tasciotti E, Liu X, Bhavane R, Plant K, Leonard AD, Price BK, Cheng MM, Decuzzi P, Tour JM, Robertson F, Ferrari M. Mesoporous silicon particles as a multistage delivery system for imaging and therapeutic applications. Nat Nanotechnol 3, 151-7 (2008). [2] Martinez JO, Evangelopoulos M, Karun V, Shegog E, Wang JA, Boada C, Liu X, Ferrari M, Tasciotti E. The effect of multistage nanovector targeting of VEGFR2 positive tumor endothelia on cell adhesion and local payload accumulation. Biomaterials 35, 9824-32 (2014). [3] Martinez JO, Evangelopoulos M, Bhavane R, Acciardo S, Salvatore F, Liu X, Ferrari M, Tasciotti E. Multistage Nanovectors Enhance the Delivery of Free and Encapsulated Drugs. Drug Targets [Epub ahead of print] (2014). [4] Parodi A, Quattrocchi N, van de Ven AL, Chiappini C, Evangelopoulos M, Martinez JO, Brown BS, Khaled SZ, Yazdi IK, Enzo MV, Isenhart L, Ferrari M, Tasciotti E. Synthetic nanoparticles functionalized with biomimetic leukocyte membranes possess cell-like functions. Nat Nanotechnol 8,61-8 (2013). [5] Parodi A, Corbo C, Cevenini A, Molinaro R, Palomba R, Pandolfi L, Agostini M, Salvatore F, Tasciotti E. Enabling cytoplasmic delivery and organelle targeting by surface modification of nanocarriers. Nanomedicine UK 10,1923-40 (2015). [6] Corbo C, Parodi A, Evangelopoulos M, Engler DA, Matsunami RK, Engler AC, Molinaro R, Scaria S, Salvatore F, Tasciotti E. Proteomic profiling of a biomimetic drug delivery platform. Curr Drug Targets [Epub ahead of print] (2014). [7] Palomba R, Parodi A, Evangelopoulos M, Acciardo S, Corbo C, De Rosa E, Yazdi I, Scaria S, Salvatore F, Tasciotti E. A leukolike vector increase the permeability of tumor vasculature. Submitted. [8] D'Apolito R, Tomaiuolo G, Taraballi F, Minardi S, Kirui D, Liu X, Cevenini A, Palomba R, Ferrari M, Salvatore F, Tasciotti E, Guido S. Red blood cells affect the margination of microparticles in synthetic microcapillaries and intravital microcirculation as a function of their size and shape. J Control Release [Epub ahead of print] (2015). [9] Parodi A, Haddix SG, Taghipour N, Scaria S, Taraballi F, Cevenini A, Yazdi IK, Corbo C, Palomba R, Khaled SZ, Martinez JO, Brown BS, Isenhart L, Tasciotti E. Bromelain surface modification increases the diffusion of silica nanoparticles in the tumor extracellular matrix. ACS Nano 8,9874-83 (2014). [10] Khaled SZ, Cevenini A, Yazdi IK, Parodi A, Evangelopoulos M, Corbo C, Scaria S, Hu Y, Chiappini C, Haddix SG, Corradetti B, Salvatore F, Tasciotti E. One-pot synthesis of pH-responsive hybrid nanogel particles for the intracellular delivery of small interfering RNA. Under review for pubblication in Biomaterials. [11] Corbo C, Molinaro R, Parodi A, Toldeno Furman NE, Salvatore F, Tasciotti E. The impact of the protein corona on nanoparticles and implications for toxicity, immunotoxicity and target drug delivery. Under review for publication in Nanomedicine UK.