Whole-cell SELEX aptamer-functionalised poly(ethyleneglycol)-

poly(ε-caprolactone) nanoparticles for enhanced targeted glioblastoma

therapy

Glioma was the most commonly diagnosed brain cancer at the time,
accounting for approximately 45%e50% of primary brain cancer cases [2]

Among all the types of cancer glioma was one of the most commonly diagnosed
brain cancer ,normally 45% to 50% cases were of glioma.[2]

. The poor prognosis associated with GBM is largely because of the poorly
defined tumour border of GBM, incomplete resection of the tumour and
tumour recurrence after surgical resection [4]. Chemotherapy is an
essential part of GBM therapy, but it is mainly ineffective due to the
small amount of drug accumulation in GBM cells, drug resistance and
serious side effects [4]

For the treatment of glioma chemotherapy is normally done but there are
many problems assosiated with chemotherapy includes ineffectiveness due
to low ammount of drug accumulation in GBM cells , many side effects and
drug resistance.[4]

There are several targeting strategies including receptor-mediated,

carrier-mediated, adsportive-mediated targeting and other physical
methods [5].

There are a number of targeting schemes which includes

adsportive-mediated ,receptor-mediated,carrier-mediated targeting and
other physical strategies.[5]

Aptamers are single stranded DNA, RNA or modified nucleic acids that can
specifically bind to their targets [6].

By definition aptamers are single stranded DNA RNA or altered nucliec

acids which have the ability to bind with a particular target.[6]

Specific aptamers can be selected from a library of random sequences

through systematic evolution of ligands by exponential enrichment (SELEX)
[6,7].
Particular aptamers can be chosen from a library of random sequence with
the help of systematic evolution of ligands by exponential enrichment
(SELEX)[6,7]

Docetaxel (DTX), a taxane derivative similar to paclitaxel, is an

inhibitor of microtubule depolymerisation [8] that is widely used for the
treatment of many cancers including brain cancer [9].

GMT8 aptamers were synthesised by the Sangon Biotech Co., Ltd (Shanghai,
China). Methoxy poly(ethyleneglycol)-poly(ε-caprolactone) (MPEG-PCL,
Mw: 3k15k) and R-carboxyl poly(ethylene glycol)-poly(ε-caprolactone)
(HOOC-PEG-PCL, Mw: 3.4k-15k) were synthesised as previously described
[10].

Targeted delivery by systems functionalised with specific ligands is a

promising strategy for the carriers, as they could have both the
anti-cancer property of the drugs and the active targeting effect of the
ligand at the same time. Several strategies were developed for this
purpose, which has shown promising results [26e28].

Carriers are the main components of targeted delivery systems. Many

carriers have been developed for drug delivery, including liposomes,
micelles, nanoparticles, polymersomes and dendrimers [29e33].

Patient safety, which is what researchers unceasingly aspire to achieve,

is a critical requirement for carriers [34].

Targeting ligands are the most important part of a targeted drug delivery
system. Many researchers focus on the newly emerged ligands for specific
receptors that are overexpressed in the target cells or on the redesign
of existing ligands, such as lactoferrin [36], angiopep-2 derived from
aprotinin [36], a 16-residue peptide (CDX) derived from candoxin [31] and
a 30-amino-acid peptide (leptin30) derived from an endogenic
hormone-leptin [37].

However, these ligands, mostly proteins or peptides, are related to

immunogenicity and competition with endogenous ligands, which may reduce
their targeting effect. Several methods have been established to select
better targeting ligands, including SELEX and phage display [6,38].

These methods can select specific short sequences of peptides or DNA from
a library that rules out the endogenous effect and immunogenicity; RGD
was the most successful example that has been demonstrated by several
groups [19,39e42].

. GMT8 is a DNA aptamer selected from a library by whole-cell SELEX against

glioma cells [7]. GMT8 aptamers can bind with high specificity to U87 cells,
which is consistent with our results showing that GMT8 aptamers
significantly enhance the uptake of ApNP compared with unmodified NP by
U87 cells. These results suggest that GMT8 aptamers could serve as an
effective ligand for GBM therapy.[article]

We established a GBM-targeted delivery system utilising GMT8 aptamers as

the targeting ligand. In vitro cellular uptake and tumour spheroid uptake
demonstrated the target effect of ApNP; cell apoptosis and tumour spheroid
growth inhibition attested to its improved anti-tumour cell growth effect.
In vivo imaging and antiGBM experiments further demonstrated that ApNP
could effectively target GBM and have an anti-GBM effect.[article]

Reference

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[6] Shangguan D, Li Y, Tang Z, Cao ZC, Chen HW, Mallikaratchy P, et al.

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[7] Bayrac AT, Sefah K, Parekh P, Bayrac C, Gulbakan B, Oktem HA, et al.
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acid as a ligand of polyethylenimine/DNA nanoparticles for targeted gene
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[27] Pang Z, Feng L, Hua R, Chen J, Gao H, Pan S, et al.

Lactoferrin-conjugated biodegradable polymersome holding doxorubicin
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[28] Pang Z, Gao H, Yu Y, Guo L, Chen J, Pan S, et al. Enhanced

intracellular delivery and chemotherapy for glioma rats by
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[30] Tang R, Ji W, Panus D, Palumbo RN, Wang C. Block copolymer micelles

with acid-labile ortho ester side-chains: Synthesis, characterization,
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[31] Zhan C, Li B, Hu L, Wei X, Feng L, Fu W, et al. Micelle-based

brain-targeted drug delivery enabled by a nicotine acetylcholine receptor
ligand. Angew Chem Int Ed Engl; 2011.

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and doxorubicin co-delivery targeting to tumor using peptide-conjugated
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[37] Liu Y, Li J, Shao K, Huang R, Ye L, Lou J, et al. A leptin derived

30-amino-acid peptide modified pegylated poly-L-lysine dendrigraft for
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[38] Pasqualini R, Ruoslahti E. Organ targeting in vivo using phage

display peptide libraries. Nature 1996;380(6572):364e6.

[42] Tyler MA, Ulasov IV, Borovjagin A, Sonabend AM, Khramtsov A, Han Y,
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poly(ethylene glycol)-co-poly(lactic acid) micelle enhances paclitaxel
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Activatable Fluorescence/MRI Bimodal Platform for Tumor Cell Imaging via
MnO2 Nanosheet−Aptamer Nanoprobe

We report a novel dual-activatable fluorescence/MRI strategy for bimodal

cancer imaging using a nanoprobe based on the physisorption of Cy5-labeled
aptamers on redoxable MnO2 nanosheets (Scheme 1). To date, various
materials based on Mn, such as MnCl2, 12 manganese chelates,13 MnO nano
particles,14 and hybrid nanomaterials,15 have been used as contrast
agents.

A novel dual-activatable fluorescence/MRI bimodal platform is designed

for tumor cell imaging by using a redoxable manganese dioxide (MnO2)
nanosheet− aptamer nanoprobe. The redoxable MnO2 nanosheet acts as a DNA
nanocarrier, fluorescence quencher, and intracellular glutathione
(GSH)-activated MRI contrast agent. In the absence of target cells,
neither fluorescence signaling nor MRI contrast of the nanoprobe is
activated. In the presence of target cells, the binding of aptamers to
their targets weakens the adsorption of aptamers on the MnO2 nanosheets,
causing partial fluorescence recovery, illuminating the target cells, and
also facilitating the endocytosis of nanoprobes into target cells. After
endocytosis, the reduction of MnO2 nanosheets by GSH further activates
the fluorescence signals and generates large amounts of Mn2+ ions suitable
for MRI. This platform should facilitate the development of various
dualactivatable fluorescence/MRI bimodalities for use in cells or in
vivo.[16]

Reference

[12]Aoki, I.; Takahashi, Y.; Chuang, K. H.; Silva, A. C.; Igarashi, T.;
Tanaka, C.; Childs, R. W.; Koretsky, A. P. NMR Biomed. 2006, 19, 50.

[13] (a) Troughton, J. S.; Greenfield, M. T.; Greenwood, J. M.; Dumas,

S.; Wiethoff, A. J.; Wang, J.; Spiller, M.; McMurry, T. J.; Caravan, P.
Inorg. Chem. 2004, 43, 6313. (b) Loving, G. S.; Mukherjee, S.; Caravan,
P. J. Am. Chem. Soc. 2013, 135, 4620.

[14] Kim, T.; Momin, E.; Choi, J.; Yuan, K.; Zaidi, H.; Kim, J.; Park,
M.; Lee, N.; McMahon, M. T.; Quinones-Hinojosa, A.; Bulte, J. W.; Hyeon,
T.; Gilad, A. A. J. Am. Chem. Soc. 2011, 133, 2955

[15] Schladt, T. D.; Shukoor, M. I.; Schneider, K.; Tahir, M. N.; Natalio,
F.; Ament, I.; Becker, J.; Jochum, F. D.; Weber, S.; Kö hler, O.; Theato,
P.; Schreiber, L. M.; Sö nnichsen, C.; Schrö der, H. C.; Müller, W.
E.; Tremel, W. Angew. Chem., Int. Ed. 2010, 49, 3976

[16] Activatable Fluorescence/MRI Bimodal Platform for Tumor Cell Imaging

via MnO2 Nanosheet−Aptamer Nanoprobe
We described the development of integrin-targeting magnetic
nanoparticles (Aptavb3-MNPs) by modifying integrin avb3-targeting
aptamer (Aptavb3) to produce a particle with high ability to detect
integrin avb3 expression in cancer cells during angiogenesis using MR
imaging. Aptavb3-MNPs exhibited no cytotoxicity and excellent
tumor-targeting detection ability with high magnetic sensitivity,
thereby demonstrating the superb performance of Aptavb3 as a targeting
vector. Furthermore, the incorporation of other diagnostic agents and
therapeutic agents in Aptavb3-MNPs might provide more accurate tumor
diagnosis and therapy.[1]

Refernce

[1] Aptamer-conjugated magnetic nanoparticles enable efficient targeted

detection of integrin avb3 via magnetic resonance imaging

MUC1 aptamer conjugated to chitosan nanoparticles, an efficient targeted

carrier designed for anticancer SN38 delivery
Aptamer-Conjugated Nanoparticles for Cancer Cell Detection

The development of aptamers for molecular recognition addresses several

of these issues: aptamers possess a low molecular weight, exhibit easy
and reproducible synthesis, easy modification,6 fast tissue penetration,
low toxicity or immunogenicity,7,8 easy storage, and high binding
affinity and selectivity.9,10

The panel of aptamers generated is ideal for profiling and personalized

medicine, as different aptamers recognize different molecular targets on
the surface of the cell which could be subsequently be correlated to
advancing patient and treatment outcomes. Panels of aptamers targeting
leukemia cell lines11, lung cancer cell lines12, liver cancer cell lines13,
and lymphoma cell lines14 have previously been described.

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Aptamer-conjugated nanomaterials and their applications

By combining the inherent features of nanomaterials with the specific

recognition ability of aptamers, a range of nanomaterial- aptamer
conjugates have proven their utility in multiple areas. 6,7

Aptamers are single-stranded oligonucleotides, DNA or RNA, with the

ability to bind to non-nucleic acid target molecules, such as peptides,
proteins, drugs, organic and inorganic molecules, or even whole cells,
with high affinity and specificity.8-12 They are isolated and chemically
synthesized from 1012 –1015 combinatorial oligonucleotide libraries by
a process known as in vitro systematic evolution of ligands by exponential
enrichment (SELEX). 13,14 Over multiple rounds of selection (generally
6–18 rounds), quite large populations (>1013 different sequences) can
be sieved, and the few nucleic acid species with specificity to the target
can be isolated[15]

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