Page 26 - ASME_NEMB_2016_Program

Technical Program TRACK 2

Roberto Molinaro, Houston Methodist Research Institute, Houston, diseases) but share the same inflammatory background.
TX, United States, Dickson Kirui, Naval Medical Research Unit
San Antonio, San Antonio, TX, United States, Jonathan Martinez, 12:20pm Immunological Impact of Membrane-derived Biomi-
Michael Evangelopoulos, Enrica De Rosa, Claudia Corbo, Ales- metic Nanoparticles
sandro Parodi, Houston Methodist Research Institute, Houston, TX,
United States, Ennio Tasciotti, The Methodist Hospital Research Technical Presentation. NEMB2016-6047
Institute, Houston, TX, United States
Michael Evangelopoulos, Michael Evangelopoulos, Alessandro
A primary directive in nanotechnology is to develop drug delivery platforms Parodi, Claudia Corbo, Jonathan Martinez, Mauro Ferrari, Hous-
that effectively reduce systemic toxicity while retaining the activity of the ton Methodist Research Institute, Houston, TX, United States, Ennio
drug, thereby increasing the therapeutic index. The ideal delivery platform Tasciotti, The Methodist Hospital Research Institute, Houston, TX,
should: i) modulate drug bioavailability and pharmacokinetics, ii) protect United States
the payload from degradation/clearance in biological environments, and iii)
favor the biodistribution and targeting at the site of interest. These proper-
ties are particularly relevant for the delivery of drugs characterized by high Recent discoveries have demonstrated that biomimicry can be employed to
toxicity (chemotherapeutics) and of compounds with poor stability in harsh bestow synthetic carriers with surface modifications derived from biological
biological environments (siRNA). Several organic (lipids, polymers) and in- components, thereby providing robust physiological tolerance and biolog-
ical targeting. With the recent popularity of biomimetic delivery platforms,
organic (silicon, silica, gold and iron oxide) materials were chosen for their
biocompatibility and have been manipulated at the micro and nano scale to the evaluation of carrier biocompatibility continues to remain a major com-
synthesize a plethora of drug delivery systems. For both types of materials, ponent for the successful translation into the clinic. Our group previously
the addition of proper surface modifications represents a key step for the demonstrated cellular membranes of circulating leukocytes could be easily
achievement of high therapeutic efficacy. A majority of currently available isolated and transferred onto the surface of synthetic nanocarriers while
maintaining key biological functions. These carriers, referred to as Leuko-
nanotherapeutics must be equipped with a coating of hydrophilic polymers,
e.g. polyethylene glycol, to prevent surface interaction with opsonizing like Vectors (LLV) [1], are composed of a loadable, biodegradable [2], and
agents and delay early sequestration by the cells of the mononuclear biocompatible [3] multistage nanovector (MSV) [4] core with a proteolipid
phagocytic system (MPS). Similarly, to achieve selective delivery to the dis- outer shell purified from leukocyte cell membrane (CM). This outer layer was
eased tissue, it is necessary to functionalize the surface of the carrier with shown to retain membrane proteins [5] that are critical for increased adhe-
sion towards inflamed endothelium while also providing prolonged circula-
targeting molecules, e.g. aptamers, antibodies, folate, transferrin. In the ef-
forts to engineer such materials, the scientific community was compelled to tion time. Despite this, while the functionalization of drug delivery vectors
challenge the extraordinary ability of our body to recognize, label, seques- with cellular materials have been demonstrated to provide synthetic parti-
ter, and clear foreign objects. In this scenario, bio-inspired approaches have cles with unique biological properties, current literature lacks a comprehen-
emerged as a one-step solution to simultaneously evade MPS and negotiate sive analysis on the biological impact of these new platforms; in particular,
for what pertains the systemic inflammation and immune response that may
the transport across various biological barriers. In particular, the physiology
of circulating immune cells provides a valuable example for the develop- arise following particle administration. Herein, we comparatively analyzed
ment of injectable carriers. Leukocytes freely circulate in the bloodstream unmodified multistage nanovectors with particles functionalized with murine
and accumulate in the diseased tissue through the selective interaction with and human leukocyte cellular membranes and the immunological effects
inflamed vasculature. that may arise in vitro and in vivo. Previously, LLV demonstrated an active
avoidance of opsonization and phagocytosis, in addition to superior target-
ing of inflammation and prolonged circulation. In this work, we performed a
In this study, we investigated how leukocyte membranes could be manipu-
lated and exploited as a proteolipid material to formulate a new generation comprehensive evaluation of the importance of the source of cellular mem-
of biomimetic drug delivery systems. These biomimetic nanovesicles (BVs) brane in increasing their systemic tolerance and minimizing an inflammatory
are a novel hybrid formulation that combines the leukocyte’s ability to es- response. Time-lapse microscopy revealed LLV developed using a cellular
coating derived from a syngeneic murine source resulted in the avoidance
cape immune surveillance, target the inflamed vasculature, and cross the
endothelial barrier with the liposome’s capacity to load, retain and release of macrophage uptake, decreased liver accumulation and no impact on he-
a cadre of different payloads in a controlled fashion. Particles’ ability to es- patic function. As biomimicry continues to develop, this work demonstrates
cape the MPS and target the inflamed endothelium has been investigated the necessity to consider the source of biological material in the develop-
both in vitro and in vivo. The in vitro exposure of BVs to macrophages was ment of drug delivery carriers. Our findings indicate the source of membrane
is critical in inhibiting cellular internalization and rapid clearance of the parti-
characterized by a significant reduction in phagocytosis, confirming the
stealth properties of the biomimetic proteolipid material. When systemically cles in vitro and in vivo, respectively, although less so in triggering an acute
injected, BVs exhibited a significant reduction in spleen and liver uptake at inflammatory response.
1 hour (10 and 50 fold decrease, respectively), compared to bare liposomes.
In addition, blood circulation studies demonstrated a 5-fold increase in pro- [1] Parodi A, Quattrocchi N, van de Ven AL, Chiappini C, Evangelopoulos M,
Martinez JO, et al. Synthetic nanoparticles functionalized with biomimetic
longed circulation. Using an in vitro flow chamber we tested the adhesion
of BVs under physiologically relevant shear stresses. Compared to conven- leukocyte membranes possess cell-like functions. Nature nanotechnology.
tional liposomes, BVs retained the targeting features of leukocytes and pref- 2013;8:61-8.
erentially recognized the inflamed endothelium. A similar comparison was [2] Martinez JO, Chiappini C, Ziemys A, Faust AM, Kojic M, Liu X, et al. En-
evaluated in vivo using intravital microscopy (IVM) on lipopolysaccharide-in- gineering multi-stage nanovectors for controlled degradation and tunable
release kinetics. Biomaterials. 2013;34:8469-77.
duced and tumor-associated models of inflammation. In both cases, BVs
exhibited an increased adhesion toward tumor-associated vasculature at 1 [3] Martinez JO, Boada C, Yazdi IK, Evangelopoulos M, Brown BS, Liu X, et
and 24 h after injection. Lastly, IVM analysis of the adhesion dynamics of BVs al. Short and long term, in vitro and in vivo correlations of cellular and tissue
to inflamed tumor endothelium revealed that while bare liposomes uniformly responses to mesoporous silicon nanovectors. Small. 2013;9:1722-33.
distributed across the vessel section, BVs exhibited substantial accumulation [4] Tasciotti E, Liu X, Bhavane R, Plant K, Leonard AD, Price BK, et al. Meso-
porous silicon particles as a multistage delivery system for imaging and ther-
at the vessel walls. This behavior remarkably mimics the innate ability of leu-
kocytes to selectively adhere to inflamed vessels prior to trans-endothelial apeutic applications. Nature nanotechnology. 2008;3:151-7.
migration. In conclusion, here we showed a proof of concept demonstration [5] Corbo C, Parodi A, Evangelopoulos M, Engler DA, Matsunami RK, Engler
of the use of proteolipid materials isolated from living cells to impart bio- AC, et al. Proteomic profiling of a biomimetic drug delivery platform. Current
logical properties and functions to synthetic liposome-like vesicles. Finally, drug targets. 2014.
26 this approach can be used as a technological platform for diagnostic and
therapeutic applications suitable to a broad range of disorders that have low 12:40pm Nanochannel platforms for tightly controlled thera-
therapeutic alternatives (e.g., rheumatoid arthritis, cancer, inflamed bowel peutic delivery and cell transplantation