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Sequential delivery of an anticancer drug and combined immunomodulatory nanoparticles for efficient chemoimmunotherapy.

Heo MB, Kim SY, Yun WS, Lim YT - Int J Nanomedicine (2015)

Bottom Line: However, CpG ODNs also induced the secretion of interleukin-10 (IL-10) that reduces the Th1 response and enhances the T helper 2 (Th2) response.Treatment of BMDCs with both types of PLGA NPs increased the Th1/Th2 cytokine (IL-12/IL-10) expression ratio, which is important for the effective induction of an antitumor immune response.After primary injection with the HA/PTX complex, the tumor-associated antigen was generated and taken up by tumor-recruited BMDCs.

View Article: PubMed Central - PubMed

Affiliation: SKKU Advanced Institute of Nanotechnology (SAINT), School of Chemical Engineering, Sungkyunkwan University, Suwon, Republic of Korea ; Center for Nanosafety Metrology, Division of Convergence Technology, Korea Research Institute of Standards and Science, Daejeon, Republic of Korea.

ABSTRACT
Chemoimmunotherapy combines chemotherapy based on anticancer drugs with immunotherapy based on immune activators to eliminate or inhibit the growth of cancer cells. In this study, water-insoluble paclitaxel (PTX) was dispersed in water using hyaluronic acid (HA) to generate a tumor-associated antigen in the tumor microenvironment. Cytosine-phosphate-guanosine oligodeoxynucleotides (CpG ODNs) were used to enhance the T helper (Th) 1 immune response. However, CpG ODNs also induced the secretion of interleukin-10 (IL-10) that reduces the Th1 response and enhances the T helper 2 (Th2) response. Therefore, RNA interference was used to downregulate IL-10 secretion from bone marrow-derived den-dritic cells (BMDCs). For the combined immunomodulation of BMDCs, we fabricated two types of poly(lactic-co-glycolic acid) (PLGA) nanoparticles (NPs) containing CpG ODNs to activate BMDCs via Toll-like receptor 9 (CpG ODN-encapsulated PLGA NPs, PCNs) or a small interfering RNA to silence IL-10 (IL-10 small interfering RNA-encapsulated PLGA NPs, PINs). Treatment of BMDCs with both types of PLGA NPs increased the Th1/Th2 cytokine (IL-12/IL-10) expression ratio, which is important for the effective induction of an antitumor immune response. After primary injection with the HA/PTX complex, the tumor-associated antigen was generated and taken up by tumor-recruited BMDCs. After a secondary injection with immunomodulating PCNs and PINs, the BMDCs became activated and migrated to the tumor-draining lymph nodes. As a result, the combination of chemotherapy using the HA/PTX complex and immunotherapy using PCNs and PINs not only efficiently inhibited tumor growth but also increased the animal survival rate. Taken together, our results suggest that the sequential treatment of cancer cells with a chemotherapeutic agent and immunomodulatory nanomaterials represents a promising strategy for efficient cancer therapy.

No MeSH data available.


Related in: MedlinePlus

In vitro gene silencing effect of PLGA NPs.Notes: BMDCs (2×106 cells) were unmanipulated (no treatment) or transfected with empty PLGA NPs (mock) or with 100 nM, 300 nM, or 500 nM PINs for 24 hours. Then, the DCs were stimulated with 5 µg/mL PCNs for 24 hours. The cells were assessed for IL-10 mRNA expression via (A) RT-PCR and (B) real-time PCR (*P<0.05 vs the Mock group).Abbreviations: PLGA, poly(lactic-co-glycolic acid); NP, nanoparticle; BMDC, bone marrow-derived dendritic cell; IL, interleukin; mRNA, messenger RNA; RT-PCR, reverse transcriptase–polymerase chain reaction; siRNA, small interfering RNA; GAPDH, glyceraldehyde 3-phosphate dehydrogenase.
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f4-ijn-10-5981: In vitro gene silencing effect of PLGA NPs.Notes: BMDCs (2×106 cells) were unmanipulated (no treatment) or transfected with empty PLGA NPs (mock) or with 100 nM, 300 nM, or 500 nM PINs for 24 hours. Then, the DCs were stimulated with 5 µg/mL PCNs for 24 hours. The cells were assessed for IL-10 mRNA expression via (A) RT-PCR and (B) real-time PCR (*P<0.05 vs the Mock group).Abbreviations: PLGA, poly(lactic-co-glycolic acid); NP, nanoparticle; BMDC, bone marrow-derived dendritic cell; IL, interleukin; mRNA, messenger RNA; RT-PCR, reverse transcriptase–polymerase chain reaction; siRNA, small interfering RNA; GAPDH, glyceraldehyde 3-phosphate dehydrogenase.

Mentions: The specificity of siRNA for gene inhibition was investigated after transfecting BMDCs with PINs containing various concentrations of IL-10 siRNA. Immature BMDCs were transfected with empty PLGA NPs (mock) or with PINs encapsulating 100 nM, 300 nM, or 500 nM siRNA. After 24 hours, the BMDCs were matured via the addition of 5 µg/mL PCNs encapsulating CpG ODNs. After maturation in CpG ODNs for 24 hours, the in vitro gene-silencing effects of PINs were determined via RT-PCR (Figure 4A) and real-time PCR (Figure 4B) analyses. The level of knock down was dependent on the treated concentration of siRNA (100–500 nM). The PINs containing 500 nM IL-10 siRNA reduced the IL-10 gene expression level bŷ70% compared with the mock treatment.


Sequential delivery of an anticancer drug and combined immunomodulatory nanoparticles for efficient chemoimmunotherapy.

Heo MB, Kim SY, Yun WS, Lim YT - Int J Nanomedicine (2015)

In vitro gene silencing effect of PLGA NPs.Notes: BMDCs (2×106 cells) were unmanipulated (no treatment) or transfected with empty PLGA NPs (mock) or with 100 nM, 300 nM, or 500 nM PINs for 24 hours. Then, the DCs were stimulated with 5 µg/mL PCNs for 24 hours. The cells were assessed for IL-10 mRNA expression via (A) RT-PCR and (B) real-time PCR (*P<0.05 vs the Mock group).Abbreviations: PLGA, poly(lactic-co-glycolic acid); NP, nanoparticle; BMDC, bone marrow-derived dendritic cell; IL, interleukin; mRNA, messenger RNA; RT-PCR, reverse transcriptase–polymerase chain reaction; siRNA, small interfering RNA; GAPDH, glyceraldehyde 3-phosphate dehydrogenase.
© Copyright Policy
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4590313&req=5

f4-ijn-10-5981: In vitro gene silencing effect of PLGA NPs.Notes: BMDCs (2×106 cells) were unmanipulated (no treatment) or transfected with empty PLGA NPs (mock) or with 100 nM, 300 nM, or 500 nM PINs for 24 hours. Then, the DCs were stimulated with 5 µg/mL PCNs for 24 hours. The cells were assessed for IL-10 mRNA expression via (A) RT-PCR and (B) real-time PCR (*P<0.05 vs the Mock group).Abbreviations: PLGA, poly(lactic-co-glycolic acid); NP, nanoparticle; BMDC, bone marrow-derived dendritic cell; IL, interleukin; mRNA, messenger RNA; RT-PCR, reverse transcriptase–polymerase chain reaction; siRNA, small interfering RNA; GAPDH, glyceraldehyde 3-phosphate dehydrogenase.
Mentions: The specificity of siRNA for gene inhibition was investigated after transfecting BMDCs with PINs containing various concentrations of IL-10 siRNA. Immature BMDCs were transfected with empty PLGA NPs (mock) or with PINs encapsulating 100 nM, 300 nM, or 500 nM siRNA. After 24 hours, the BMDCs were matured via the addition of 5 µg/mL PCNs encapsulating CpG ODNs. After maturation in CpG ODNs for 24 hours, the in vitro gene-silencing effects of PINs were determined via RT-PCR (Figure 4A) and real-time PCR (Figure 4B) analyses. The level of knock down was dependent on the treated concentration of siRNA (100–500 nM). The PINs containing 500 nM IL-10 siRNA reduced the IL-10 gene expression level bŷ70% compared with the mock treatment.

Bottom Line: However, CpG ODNs also induced the secretion of interleukin-10 (IL-10) that reduces the Th1 response and enhances the T helper 2 (Th2) response.Treatment of BMDCs with both types of PLGA NPs increased the Th1/Th2 cytokine (IL-12/IL-10) expression ratio, which is important for the effective induction of an antitumor immune response.After primary injection with the HA/PTX complex, the tumor-associated antigen was generated and taken up by tumor-recruited BMDCs.

View Article: PubMed Central - PubMed

Affiliation: SKKU Advanced Institute of Nanotechnology (SAINT), School of Chemical Engineering, Sungkyunkwan University, Suwon, Republic of Korea ; Center for Nanosafety Metrology, Division of Convergence Technology, Korea Research Institute of Standards and Science, Daejeon, Republic of Korea.

ABSTRACT
Chemoimmunotherapy combines chemotherapy based on anticancer drugs with immunotherapy based on immune activators to eliminate or inhibit the growth of cancer cells. In this study, water-insoluble paclitaxel (PTX) was dispersed in water using hyaluronic acid (HA) to generate a tumor-associated antigen in the tumor microenvironment. Cytosine-phosphate-guanosine oligodeoxynucleotides (CpG ODNs) were used to enhance the T helper (Th) 1 immune response. However, CpG ODNs also induced the secretion of interleukin-10 (IL-10) that reduces the Th1 response and enhances the T helper 2 (Th2) response. Therefore, RNA interference was used to downregulate IL-10 secretion from bone marrow-derived den-dritic cells (BMDCs). For the combined immunomodulation of BMDCs, we fabricated two types of poly(lactic-co-glycolic acid) (PLGA) nanoparticles (NPs) containing CpG ODNs to activate BMDCs via Toll-like receptor 9 (CpG ODN-encapsulated PLGA NPs, PCNs) or a small interfering RNA to silence IL-10 (IL-10 small interfering RNA-encapsulated PLGA NPs, PINs). Treatment of BMDCs with both types of PLGA NPs increased the Th1/Th2 cytokine (IL-12/IL-10) expression ratio, which is important for the effective induction of an antitumor immune response. After primary injection with the HA/PTX complex, the tumor-associated antigen was generated and taken up by tumor-recruited BMDCs. After a secondary injection with immunomodulating PCNs and PINs, the BMDCs became activated and migrated to the tumor-draining lymph nodes. As a result, the combination of chemotherapy using the HA/PTX complex and immunotherapy using PCNs and PINs not only efficiently inhibited tumor growth but also increased the animal survival rate. Taken together, our results suggest that the sequential treatment of cancer cells with a chemotherapeutic agent and immunomodulatory nanomaterials represents a promising strategy for efficient cancer therapy.

No MeSH data available.


Related in: MedlinePlus