EMulate Therapeutics has developed a system for emulating the effects of solvated molecules via their magnetic field recordings. Recordings of magnetic field emissions of select small inhibitor RNAs (siRNAs;murine tar...EMulate Therapeutics has developed a system for emulating the effects of solvated molecules via their magnetic field recordings. Recordings of magnetic field emissions of select small inhibitor RNAs (siRNAs;murine targeting CTLA-4 and murine targeting PD-1) were tested on C57Bl/6 mice implanted subcutaneously with the GL261 murine tumor cell line. A signal composed of concatenated recordings of siRNA molecules targeting the murine CTLA-4 and PD-1 receptors (labeled A2) was used in immune competent C57Bl/6 mice. The mice were flank implanted with the murine glioblastoma cell line GL261. Mice were exposed to the signal continuously (24 hours a day) until tumor volumes reached the designated volume limit. Tumors were excised and analyzed via PAGE/Western blot for the expression of CTLA-4, PD-1, Ki67, Caspase 3, CD4 and CD8. Terminal blood draws were used for CBCs. We report the down regulation of the checkpoint inhibitors CTLA-4 in the exposed mice. Significant tumor volume reduction was observed in mice exposed to the siRNA signal compared to control mice;no adverse events were recorded. Cell blood counts (CBC) and protein expression patterns were observed to correlate with the expected function of protein expression inhibition of the targets.展开更多
BACKGROUND Less than 0.5%of intravenously injected drugs reach tumors,contributing to side effects.To limit damage to healthy cells,various delivery vectors have been formulated;yet,previously developed vectors suffer...BACKGROUND Less than 0.5%of intravenously injected drugs reach tumors,contributing to side effects.To limit damage to healthy cells,various delivery vectors have been formulated;yet,previously developed vectors suffer from poor penetration into solid tumors.This issue was resolved by the discovery of HN-1 peptide isolated via biopanning a phage-display library.HN-1 targets human head and neck squamous cell carcinoma(HNSCC)(breast,thyroid;potentially lung,cervix,uterine,colon cancer),translocates across the cell membrane,and efficiently infiltrates solid tumors.HN-1 peptide has been conjugated to various anticancer drugs and imaging agents though the identity of its receptor remained enigmatic.AIM To decipher the clues that pointed to retinoblastoma(Rb)-regulated discoidindomain receptor 1 as the putative receptor for HN-1 is described.METHODS HN-1 peptide was synthesized and purified using reverse-phase highperformance liquid chromatography and gel electrophoresis.The predicted mass was confirmed by mass spectroscopy.To image the 3-dimensional structure of HN-1 peptide,PyMOL was used.Molecular modeling was also performed with PEP-FOLD3 software via RPBS bioinformatics web portal(INSERM,France).The immunohistochemistry results of discoidin domain receptor 1(DDR1)protein were obtained from the publicly accessible database in the Human Protein Atlas portal,which contained the images of immunohistochemically labeled human cancers and the corresponding normal tissues.RESULTS The clues that led to DDR1 involved in metastasis as the putative receptor mediating HN-1 endocytosis are the following:(1)HN-1 is internalized in phosphate-buffered saline and its uptake is competitively inhibited;(2)HN-1(TSPLNIHNGQKL)exhibits similarity with a stretch of amino acids in alpha5 beta3 integrin(KLLITIHDRKEF).Aside from two identical residues(Ile-His)in the middle,the overall distribution of polar and nonpolar residues throughout the sequences is nearly identical.As HN-1 sequence lacks the Arg-Gly-Asp motif recognized by integrins,HN-1 may interact with an"integrin-like"molecule.The tertiary structure of both peptides showed similarity at the 3-dimensional level;(3)HN-1 is internalized by attached cells but not by suspended cells.As culture plates are typically coated with collagen,collagen-binding receptor(expressed by adherent but not suspended cells)may represent the receptor for HN-1;(4)DDR1 is highly expressed in head and neck cancer(or breast cancer)targeted by HN-1;(5)Upon activation by collagen,DDR1 becomes internalized and compartmentalized in endosomes consistent with the determination of’energy-dependent clathrin-mediated endocytosis’as the HN-1 entry route and the identification of HN-1 entrapped vesicles as endosomes;and(6)DDR1 is essential for the development of mammary glands consistent with the common embryonic lineage rationale used to identify breast cancer as an additional target of HN-1.In summary,collagenactivated tyrosine kinase receptor DDR1 overexpressed in HNSCC assumes a critical role in metastasis.Further studies are warranted to assess HN-1 peptide’s interaction with DDR1 and the therapeutic potential of treating metastatic cancer.Additionally,advances in delivery(conformation,endocytic mechanism,repertoire of targeted cancers of HN-1 peptide),tracking(HN-1 conjugated imaging agents),and activity(HN-1 conjugated therapeutic agents)are described.CONCLUSION The discovery of DDR1 as HN-1 peptide’s putative receptor represents a significant advance as it enables identification of metastatic cancers or clinical application of previously developed therapeutics to block metastasis.展开更多
BACKGROUND Recurrent hepatocellular carcinoma(HCC)with inferior vena cava tumor thrombus is a great challenge for oncologists and has a poor prognosis.To date,the safety and efficacy of programmed cell death ligand 1(...BACKGROUND Recurrent hepatocellular carcinoma(HCC)with inferior vena cava tumor thrombus is a great challenge for oncologists and has a poor prognosis.To date,the safety and efficacy of programmed cell death ligand 1(PD-L1)inhibitors are still unknown.CASE SUMMARY A 59-year-old male was identified as having a tumor thrombus in the inferior vena cava 3 years after surgery.The patient underwent a second surgery and adjuvant chemotherapy.However,the level of alpha-fetoprotein was elevated after 2 mo,and lung metastases and mediastinal lymph node metastases were identified.The expression of PD-L1 in HCC and inferior vena cava tumor thrombus tissues was analyzed by immunohistochemistry.Then,the patient received atezolizumab immunotherapy.The level of alpha-fetoprotein dropped to normal,the mediastinal lymph node metastases decreased in size and the lung metastases disappeared after 3 mo of immunotherapy.The patient had no signs of recurrence at 21 mo of follow-up.A 60-year-old male underwent left hepatic tumor resection,inferior vena cava incision and thrombus removal,followed by regular chemotherapy.The patient developed lung and splenic metastases after surgery.Pembrolizumab was used for six courses,and the splenic metastasis shrank,after which splenectomy was performed.The patient continued to receive pembrolizumab for thirteen courses,and the lung metastases showed no progression.A 34-year-old male was diagnosed with liver cancer with inferior vena cava tumor thrombus.The patient underwent right hepatectomy and received tislelizumab for three courses.He is still receiving immunotherapy and in good condition.CONCLUSION Anti-PD-L1 therapy in HCC patients with inferior vena cava tumor thrombus and metastasis is associated with relatively good patient outcomes.展开更多
文摘EMulate Therapeutics has developed a system for emulating the effects of solvated molecules via their magnetic field recordings. Recordings of magnetic field emissions of select small inhibitor RNAs (siRNAs;murine targeting CTLA-4 and murine targeting PD-1) were tested on C57Bl/6 mice implanted subcutaneously with the GL261 murine tumor cell line. A signal composed of concatenated recordings of siRNA molecules targeting the murine CTLA-4 and PD-1 receptors (labeled A2) was used in immune competent C57Bl/6 mice. The mice were flank implanted with the murine glioblastoma cell line GL261. Mice were exposed to the signal continuously (24 hours a day) until tumor volumes reached the designated volume limit. Tumors were excised and analyzed via PAGE/Western blot for the expression of CTLA-4, PD-1, Ki67, Caspase 3, CD4 and CD8. Terminal blood draws were used for CBCs. We report the down regulation of the checkpoint inhibitors CTLA-4 in the exposed mice. Significant tumor volume reduction was observed in mice exposed to the siRNA signal compared to control mice;no adverse events were recorded. Cell blood counts (CBC) and protein expression patterns were observed to correlate with the expected function of protein expression inhibition of the targets.
文摘BACKGROUND Less than 0.5%of intravenously injected drugs reach tumors,contributing to side effects.To limit damage to healthy cells,various delivery vectors have been formulated;yet,previously developed vectors suffer from poor penetration into solid tumors.This issue was resolved by the discovery of HN-1 peptide isolated via biopanning a phage-display library.HN-1 targets human head and neck squamous cell carcinoma(HNSCC)(breast,thyroid;potentially lung,cervix,uterine,colon cancer),translocates across the cell membrane,and efficiently infiltrates solid tumors.HN-1 peptide has been conjugated to various anticancer drugs and imaging agents though the identity of its receptor remained enigmatic.AIM To decipher the clues that pointed to retinoblastoma(Rb)-regulated discoidindomain receptor 1 as the putative receptor for HN-1 is described.METHODS HN-1 peptide was synthesized and purified using reverse-phase highperformance liquid chromatography and gel electrophoresis.The predicted mass was confirmed by mass spectroscopy.To image the 3-dimensional structure of HN-1 peptide,PyMOL was used.Molecular modeling was also performed with PEP-FOLD3 software via RPBS bioinformatics web portal(INSERM,France).The immunohistochemistry results of discoidin domain receptor 1(DDR1)protein were obtained from the publicly accessible database in the Human Protein Atlas portal,which contained the images of immunohistochemically labeled human cancers and the corresponding normal tissues.RESULTS The clues that led to DDR1 involved in metastasis as the putative receptor mediating HN-1 endocytosis are the following:(1)HN-1 is internalized in phosphate-buffered saline and its uptake is competitively inhibited;(2)HN-1(TSPLNIHNGQKL)exhibits similarity with a stretch of amino acids in alpha5 beta3 integrin(KLLITIHDRKEF).Aside from two identical residues(Ile-His)in the middle,the overall distribution of polar and nonpolar residues throughout the sequences is nearly identical.As HN-1 sequence lacks the Arg-Gly-Asp motif recognized by integrins,HN-1 may interact with an"integrin-like"molecule.The tertiary structure of both peptides showed similarity at the 3-dimensional level;(3)HN-1 is internalized by attached cells but not by suspended cells.As culture plates are typically coated with collagen,collagen-binding receptor(expressed by adherent but not suspended cells)may represent the receptor for HN-1;(4)DDR1 is highly expressed in head and neck cancer(or breast cancer)targeted by HN-1;(5)Upon activation by collagen,DDR1 becomes internalized and compartmentalized in endosomes consistent with the determination of’energy-dependent clathrin-mediated endocytosis’as the HN-1 entry route and the identification of HN-1 entrapped vesicles as endosomes;and(6)DDR1 is essential for the development of mammary glands consistent with the common embryonic lineage rationale used to identify breast cancer as an additional target of HN-1.In summary,collagenactivated tyrosine kinase receptor DDR1 overexpressed in HNSCC assumes a critical role in metastasis.Further studies are warranted to assess HN-1 peptide’s interaction with DDR1 and the therapeutic potential of treating metastatic cancer.Additionally,advances in delivery(conformation,endocytic mechanism,repertoire of targeted cancers of HN-1 peptide),tracking(HN-1 conjugated imaging agents),and activity(HN-1 conjugated therapeutic agents)are described.CONCLUSION The discovery of DDR1 as HN-1 peptide’s putative receptor represents a significant advance as it enables identification of metastatic cancers or clinical application of previously developed therapeutics to block metastasis.
基金Supported by The Special Research Foundation of the National Nature Science Foundation of China,No.81972262 and No.81972255The Guangdong Basic and Applied Basic Research Foundation,No.2018A030313645,No.2020A1515010117 and No.2016A030313840+4 种基金Key Laboratory of Malignant Tumor Molecular Mechanism and Translational Medicine of Guangzhou Bureau of Science and Information Technology,No.[2013]163the Key Laboratory of Malignant Tumor Gene Regulation and Target Therapy of Guangdong Higher Education Institutes,No.KLB09001Guangdong Science and Technology Department,No.2015B050501004Guangdong Science and Technology Department,No.2017B030314026Sun Yat-sen University Clinical Research 5010 Program,No.2018008.
文摘BACKGROUND Recurrent hepatocellular carcinoma(HCC)with inferior vena cava tumor thrombus is a great challenge for oncologists and has a poor prognosis.To date,the safety and efficacy of programmed cell death ligand 1(PD-L1)inhibitors are still unknown.CASE SUMMARY A 59-year-old male was identified as having a tumor thrombus in the inferior vena cava 3 years after surgery.The patient underwent a second surgery and adjuvant chemotherapy.However,the level of alpha-fetoprotein was elevated after 2 mo,and lung metastases and mediastinal lymph node metastases were identified.The expression of PD-L1 in HCC and inferior vena cava tumor thrombus tissues was analyzed by immunohistochemistry.Then,the patient received atezolizumab immunotherapy.The level of alpha-fetoprotein dropped to normal,the mediastinal lymph node metastases decreased in size and the lung metastases disappeared after 3 mo of immunotherapy.The patient had no signs of recurrence at 21 mo of follow-up.A 60-year-old male underwent left hepatic tumor resection,inferior vena cava incision and thrombus removal,followed by regular chemotherapy.The patient developed lung and splenic metastases after surgery.Pembrolizumab was used for six courses,and the splenic metastasis shrank,after which splenectomy was performed.The patient continued to receive pembrolizumab for thirteen courses,and the lung metastases showed no progression.A 34-year-old male was diagnosed with liver cancer with inferior vena cava tumor thrombus.The patient underwent right hepatectomy and received tislelizumab for three courses.He is still receiving immunotherapy and in good condition.CONCLUSION Anti-PD-L1 therapy in HCC patients with inferior vena cava tumor thrombus and metastasis is associated with relatively good patient outcomes.
