23 research outputs found
Image_1_scRNA-seq characterizing the heterogeneity of fibroblasts in breast cancer reveals a novel subtype SFRP4+ CAF that inhibits migration and predicts prognosis.pdf
IntroductionCancer-associated fibroblasts (CAFs) are a diverse group of cells that significantly impact the tumor microenvironment and therapeutic responses in breast cancer (BC). Despite their importance, the comprehensive profile of CAFs in BC remains to be fully elucidated.MethodsTo address this gap, we utilized single-cell RNA sequencing (scRNA-seq) to delineate the CAF landscape within 14 BC normal-tumor paired samples. We further corroborated our findings by analyzing several public datasets, thereby validating the newly identified CAF subtype. Additionally, we conducted coculture experiments with BC cells to assess the functional implications of this CAF subtype.ResultsOur scRNA-seq analysis unveiled eight distinct CAF subtypes across five tumor and six adjacent normal tissue samples. Notably, we discovered a novel subtype, designated as SFRP4+ CAFs, which was predominantly observed in normal tissues. The presence of SFRP4+ CAFs was substantiated by two independent scRNA-seq datasets and a spatial transcriptomics dataset. Functionally, SFRP4+ CAFs were found to impede BC cell migration and the epithelial-mesenchymal transition (EMT) process by secreting SFRP4, thereby modulating the WNT signaling pathway. Furthermore, we established that elevated expression levels of SFRP4+ CAF markers correlate with improved survival outcomes in BC patients, yet paradoxically, they predict a diminished response to neoadjuvant chemotherapy in cases of triple-negative breast cancer.ConclusionThis investigation sheds light on the heterogeneity of CAFs in BC and introduces a novel SFRP4+ CAF subtype that hinders BC cell migration. This discovery holds promise as a potential biomarker for refined prognostic assessment and therapeutic intervention in BC.</p
MiRNA and mRNA-Controlled Double-Cascaded Amplifying Circuit Nanosensor for Accurate Discrimination of Breast Cancers in Living Cells, Animals, and Organoids
Metastasis is the leading cause of death in patients
with breast
cancer. Detecting high-risk breast cancer, including micrometastasis,
at an early stage is vital for customizing the right and efficient
therapies. In this study, we propose an enzyme-free isothermal cascade
amplification-based DNA logic circuit in situ biomineralization nanosensor,
HDNAzyme@ZIF-8, for simultaneous imaging of multidimensional biomarkers
in live cells. Taking miR-21 and Ki-67 mRNA as the dual detection
targets achieved sensitive logic operations and molecular recognition
through the cascade hybridization chain reaction and DNAzyme. The
HDNAzyme@ZIF-8 nanosensor has the ability to accurately differentiate
breast cancer cells and their subtypes by comparing their relative
fluorescence intensities. Of note, our nanosensor can also achieve
visualization within breast cancer organoids, faithfully recapitulating
the functional characteristics of parental tumor. Overall, the combination
of these techniques offers a universal strategy for detecting cancers
with high sensitivity and holds vast potential in clinical cancer
diagnosis
Table_1_scRNA-seq characterizing the heterogeneity of fibroblasts in breast cancer reveals a novel subtype SFRP4+ CAF that inhibits migration and predicts prognosis.xlsx
IntroductionCancer-associated fibroblasts (CAFs) are a diverse group of cells that significantly impact the tumor microenvironment and therapeutic responses in breast cancer (BC). Despite their importance, the comprehensive profile of CAFs in BC remains to be fully elucidated.MethodsTo address this gap, we utilized single-cell RNA sequencing (scRNA-seq) to delineate the CAF landscape within 14 BC normal-tumor paired samples. We further corroborated our findings by analyzing several public datasets, thereby validating the newly identified CAF subtype. Additionally, we conducted coculture experiments with BC cells to assess the functional implications of this CAF subtype.ResultsOur scRNA-seq analysis unveiled eight distinct CAF subtypes across five tumor and six adjacent normal tissue samples. Notably, we discovered a novel subtype, designated as SFRP4+ CAFs, which was predominantly observed in normal tissues. The presence of SFRP4+ CAFs was substantiated by two independent scRNA-seq datasets and a spatial transcriptomics dataset. Functionally, SFRP4+ CAFs were found to impede BC cell migration and the epithelial-mesenchymal transition (EMT) process by secreting SFRP4, thereby modulating the WNT signaling pathway. Furthermore, we established that elevated expression levels of SFRP4+ CAF markers correlate with improved survival outcomes in BC patients, yet paradoxically, they predict a diminished response to neoadjuvant chemotherapy in cases of triple-negative breast cancer.ConclusionThis investigation sheds light on the heterogeneity of CAFs in BC and introduces a novel SFRP4+ CAF subtype that hinders BC cell migration. This discovery holds promise as a potential biomarker for refined prognostic assessment and therapeutic intervention in BC.</p
Knockdown of BCRP/ABCG2 expression by V-BCRPi in JAR cells using immunofluorescence analysis (×100).
<p>The results for experimental groups 1 to 8 are shown: 1: mock cells with MoAb or 2: PBS or cells infected with 3: V-BCRP1i, 4: V-BCRP1i-c, 5: V-BCRP2i, 6: V-BCRP2i-c, 7: V-BCRP3i, or 8: V-BCRP3i-c. The fluorescence intensity of cells subjected to V-BCRP3i treatment was the lowest.</p
Changes in the apoptosis rate of transplantation tumors in nude mice injected with virus and 5-FU (x±s, n = 5).
<p>*: PBS+5-FU vs. V-BCRPi+5-FU, p<0.01.</p
Tumor bodies of hairless mice after injection of JAR cancer cells infected with V-BCRPi and treatment with 5-FU.
<p>A1: Tumor body injected with PBS alone; A2: Tumor body injected with PBS and 5-FU; B1: Tumor body injected with V-BCRPi alone; and B2: Tumor body injected with V-BCRPi and 5-FU. After injecting 5-FU, the tumors in the hairless mice injected with V-BCRPi were smaller than those in the un-injected hairless mice. The anti-tumor rate was approximately a factor of 10 (<i>P</i><0.01). It was concluded that V-BCRPi increases the inhibition effects of 5-FU on tumor growth.</p
Fenobody: A Ferritin-Displayed Nanobody with High Apparent Affinity and Half-Life Extension
Nanobodies
consist of a single domain variable fragment of a camelid
heavy-chain antibody. Nanobodies have potential applications in biomedical
fields because of their simple production procedures and low cost.
Occasionally, nanobody clones of interest exhibit low affinities for
their target antigens, which, together with their short half-life
limit bioanalytical or therapeutic applications. Here, we developed
a novel platform we named fenobody, in which a nanobody developed
against H5N1 virus is displayed on the surface of ferritin in the
form of a 24mer. We constructed a fenobody by substituting the fifth
helix of ferritin with the nanobody. TEM analysis showed that nanobodies
were displayed on the surface of ferritin in the form of 6 ×
4 bundles, and that these clustered nanobodies are flexible for antigen
binding in spatial structure. Comparing fenobodies with conventional
nanobodies currently used revealed that the antigen binding apparent
affinity of anti-H5N1 fenobody was dramatically increased (∼360-fold).
Crucially, their half-life extension in a murine model was 10-fold
longer than anti-H5N1 nanobody. In addition, we found that our fenobodies
are highly expressed in Escherichia coli, and are both soluble and thermo-stable nanocages that self-assemble
as 24-polymers. In conclusion, our results demonstrate that fenobodies
have unique advantages over currently available systems for apparent
affinity enhancement and half-life extension of nanobodies. Our fenobody
system presents a suitable platform for various large-scale biotechnological
processes and should greatly facilitate the application of nanobody
technology in these areas
Residual drug volumes after infection of JAR cells with V-BCRPi according to flow cytometry analysis.
<p>The results of experimental groups 1 to 9 are shown: 1: mock cells, 2: cells with Mit, 3: cells with Mit and Ko143, or cells with Mit and infected with 4: V-BCRP1i, 5: V-BCRP2i, 6: V-BCRP3i, 7: V-BCRP1i-c, 8: V-BCRP2i-c, or 9: V-BCRP3i-c. Each residual drug volume represents the mean value of three independent experiments. *<i>P</i><0.01.</p
Knockdown of BCRP/ABCG2 expression by V-BCRPi in tumor cell bodies, as shown by Western blot analysis.
<p>1: Tumor body injected with PBS alone; 2: Tumor body injected with PBS and 5-FU; 3: Tumor body injected with V-BCRPi alone; and 4: Tumor body injected with V-BCRPi and 5-FU. It was concluded from the experimental results that the expression of <i>BCRP/ABCG2</i> in tumors injected with V-BCRPi (with 5-FU treatment) was lower than that of the un-injected tumors (with PBS and 5-FU). There was no difference among the groups injected with the various V-BCRPi retroviruses and 5-FU.</p
Changes in tumor weight and volume and the anti-tumor rate in transplantation tumors in nude mice after virus infection and drug treatment (x±s, n = 15).
<p>*: PBS+5-FU vs. V-BCRPi+5-FU, p<0.01.</p