Last updated: 2023-04-10

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Knit directory: Cardiotoxicity/

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Rmd fa2cbeb reneeisnowhere 2023-04-10 monday end
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Rmd 1444a85 reneeisnowhere 2023-04-10 update push of new data
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Rmd 0f1f1da reneeisnowhere 2023-04-10 final run analysis

This starts the documentation of the RNA-seq cardiotoxicity analysis for my manuscript

library(edgeR)#
library(limma)#
library(RColorBrewer)
library(mixOmics)
library(gridExtra)#
library(reshape2)#
#library(devtools)
library(AnnotationHub)
library(tidyverse)
library(scales)
library(biomaRt)#
library(Homo.sapiens)
library(cowplot)#
library(ggrepel)#
library(corrplot)
library(Hmisc)
 ###now we add genenames to the geneid###
 
geneid <- rownames(mymatrix) ### pulls the names we have in the counts file
genes <- select(Homo.sapiens, keys=geneid, columns=c("SYMBOL"),
  keytype="ENTREZID")
genes <- genes[!duplicated(genes$ENTREZID),]
mymatrix$genes <- genes
[1] 28395    72
[1] 14086    72



###  Vehicle 



###  Daunorubicin 



###  Doxorubicin 



###  Epirubicin 



###  Mitoxantrone 



###  Trastuzumab 

        samplenames indv         drug time RIN group       PC1      PC2
Da.1.3h MCW_RM_R_11    1 Daunorubicin   3h 9.3     1 -18.53902 62.11914
Do.1.3h MCW_RM_R_12    1  Doxorubicin   3h 9.8     2 -12.37874 73.96424
Ep.1.3h MCW_RM_R_13    1   Epirubicin   3h 9.8     3 -11.19226 66.47554
Mi.1.3h MCW_RM_R_14    1 Mitoxantrone   3h  10     4 -10.24538 73.46900
Tr.1.3h MCW_RM_R_15    1  Trastuzumab   3h 9.6     5 -12.21578 79.97646
Ve.1.3h MCW_RM_R_16    1      Vehicle   3h 9.9     6 -15.01396 76.61739
              PC3        PC4       PC5       PC6
Da.1.3h 44.793907  -4.229684  25.33474 -35.96203
Do.1.3h 24.565270  -8.522490 -19.86738 -19.00810
Ep.1.3h 33.103318  -9.146883  18.17321 -43.12911
Mi.1.3h 19.048074 -14.540036  -9.00047 -24.33769
Tr.1.3h  2.646249 -17.004856 -34.18428 -11.83222
Ve.1.3h 12.636907  -4.140009 -39.84065 -17.16481

Importance of components:
                           PC1     PC2      PC3      PC4      PC5      PC6
Standard deviation     63.9774 47.1102 34.19844 32.56486 28.23410 23.90465
Proportion of Variance  0.2906  0.1576  0.08303  0.07529  0.05659  0.04057
Cumulative Proportion   0.2906  0.4481  0.53117  0.60645  0.66304  0.70361
                            PC7      PC8      PC9     PC10     PC11     PC12
Standard deviation     21.54760 20.74762 19.52266 17.46914 16.25667 15.49063
Proportion of Variance  0.03296  0.03056  0.02706  0.02166  0.01876  0.01704
Cumulative Proportion   0.73657  0.76713  0.79419  0.81586  0.83462  0.85165
                           PC13     PC14     PC15     PC16     PC17    PC18
Standard deviation     14.02995 13.01851 11.78389 11.40612 10.15585 9.76592
Proportion of Variance  0.01397  0.01203  0.00986  0.00924  0.00732 0.00677
Cumulative Proportion   0.86563  0.87766  0.88752  0.89675  0.90408 0.91085
                          PC19    PC20    PC21   PC22    PC23    PC24    PC25
Standard deviation     8.79138 8.66072 8.09173 7.8730 7.46626 7.09040 6.61801
Proportion of Variance 0.00549 0.00533 0.00465 0.0044 0.00396 0.00357 0.00311
Cumulative Proportion  0.91633 0.92166 0.92631 0.9307 0.93466 0.93823 0.94134
                          PC26    PC27    PC28    PC29    PC30    PC31    PC32
Standard deviation     6.31712 6.05845 5.86317 5.72293 5.54398 5.49527 5.27437
Proportion of Variance 0.00283 0.00261 0.00244 0.00233 0.00218 0.00214 0.00197
Cumulative Proportion  0.94418 0.94678 0.94922 0.95155 0.95373 0.95587 0.95785
                          PC33    PC34    PC35    PC36    PC37    PC38    PC39
Standard deviation     5.18844 5.11708 4.86120 4.83415 4.71966 4.52799 4.50677
Proportion of Variance 0.00191 0.00186 0.00168 0.00166 0.00158 0.00146 0.00144
Cumulative Proportion  0.95976 0.96162 0.96330 0.96495 0.96654 0.96799 0.96943
                          PC40    PC41    PC42    PC43    PC44    PC45    PC46
Standard deviation     4.39472 4.34878 4.30170 4.23851 4.15240 4.12290 4.05711
Proportion of Variance 0.00137 0.00134 0.00131 0.00128 0.00122 0.00121 0.00117
Cumulative Proportion  0.97080 0.97215 0.97346 0.97474 0.97596 0.97717 0.97834
                          PC47   PC48    PC49    PC50    PC51    PC52    PC53
Standard deviation     4.01633 3.9421 3.90038 3.83199 3.81069 3.80580 3.76258
Proportion of Variance 0.00115 0.0011 0.00108 0.00104 0.00103 0.00103 0.00101
Cumulative Proportion  0.97948 0.9806 0.98166 0.98271 0.98374 0.98477 0.98577
                          PC54    PC55    PC56    PC57   PC58    PC59    PC60
Standard deviation     3.71747 3.69618 3.64557 3.59456 3.5595 3.51279 3.46169
Proportion of Variance 0.00098 0.00097 0.00094 0.00092 0.0009 0.00088 0.00085
Cumulative Proportion  0.98675 0.98772 0.98866 0.98958 0.9905 0.99136 0.99221
                          PC61    PC62    PC63    PC64    PC65   PC66    PC67
Standard deviation     3.45216 3.40460 3.34569 3.27664 3.21965 3.1371 3.12182
Proportion of Variance 0.00085 0.00082 0.00079 0.00076 0.00074 0.0007 0.00069
Cumulative Proportion  0.99305 0.99388 0.99467 0.99543 0.99617 0.9969 0.99756
                          PC68    PC69    PC70   PC71      PC72
Standard deviation     3.09899 3.05675 2.88458 2.6632 8.662e-14
Proportion of Variance 0.00068 0.00066 0.00059 0.0005 0.000e+00
Cumulative Proportion  0.99824 0.99891 0.99950 1.0000 1.000e+00

##correlation heatmap of counts matrix

###now to get the counts set for DEG!!

        V.DA  V.DX  V.EP  V.MT  V.TR V.DA24 V.DX24 V.EP24 V.MT24 V.TR24
Down     108     3    29    22     0   3542   3341   3111    430      0
NotSig 13569 14067 13876 14014 14086   7062   7431   7751  12964  14086
Up       409    16   181    50     0   3482   3314   3224    692      0

R version 4.2.2 (2022-10-31 ucrt)
Platform: x86_64-w64-mingw32/x64 (64-bit)
Running under: Windows 10 x64 (build 19044)

Matrix products: default

locale:
[1] LC_COLLATE=English_United States.utf8 
[2] LC_CTYPE=English_United States.utf8   
[3] LC_MONETARY=English_United States.utf8
[4] LC_NUMERIC=C                          
[5] LC_TIME=English_United States.utf8    

attached base packages:
[1] stats4    stats     graphics  grDevices utils     datasets  methods  
[8] base     

other attached packages:
 [1] Hmisc_4.8-0                            
 [2] Formula_1.2-5                          
 [3] survival_3.5-3                         
 [4] corrplot_0.92                          
 [5] ggrepel_0.9.3                          
 [6] cowplot_1.1.1                          
 [7] Homo.sapiens_1.3.1                     
 [8] TxDb.Hsapiens.UCSC.hg19.knownGene_3.2.2
 [9] org.Hs.eg.db_3.15.0                    
[10] GO.db_3.15.0                           
[11] OrganismDbi_1.38.1                     
[12] GenomicFeatures_1.48.4                 
[13] GenomicRanges_1.48.0                   
[14] GenomeInfoDb_1.32.4                    
[15] AnnotationDbi_1.58.0                   
[16] IRanges_2.30.1                         
[17] S4Vectors_0.34.0                       
[18] Biobase_2.56.0                         
[19] biomaRt_2.52.0                         
[20] scales_1.2.1                           
[21] lubridate_1.9.2                        
[22] forcats_1.0.0                          
[23] stringr_1.5.0                          
[24] dplyr_1.1.0                            
[25] purrr_1.0.1                            
[26] readr_2.1.4                            
[27] tidyr_1.3.0                            
[28] tibble_3.1.8                           
[29] tidyverse_2.0.0                        
[30] AnnotationHub_3.4.0                    
[31] BiocFileCache_2.4.0                    
[32] dbplyr_2.3.1                           
[33] BiocGenerics_0.42.0                    
[34] reshape2_1.4.4                         
[35] gridExtra_2.3                          
[36] mixOmics_6.20.0                        
[37] ggplot2_3.4.1                          
[38] lattice_0.20-45                        
[39] MASS_7.3-58.2                          
[40] RColorBrewer_1.1-3                     
[41] edgeR_3.38.4                           
[42] limma_3.52.4                           
[43] workflowr_1.7.0                        

loaded via a namespace (and not attached):
  [1] backports_1.4.1               plyr_1.8.8                   
  [3] igraph_1.4.1                  splines_4.2.2                
  [5] BiocParallel_1.30.4           digest_0.6.31                
  [7] htmltools_0.5.4               fansi_1.0.4                  
  [9] checkmate_2.1.0               magrittr_2.0.3               
 [11] memoise_2.0.1                 cluster_2.1.4                
 [13] tzdb_0.3.0                    Biostrings_2.64.1            
 [15] matrixStats_0.63.0            rARPACK_0.11-0               
 [17] timechange_0.2.0              prettyunits_1.1.1            
 [19] jpeg_0.1-10                   colorspace_2.1-0             
 [21] blob_1.2.3                    rappdirs_0.3.3               
 [23] xfun_0.37                     callr_3.7.3                  
 [25] crayon_1.5.2                  RCurl_1.98-1.10              
 [27] jsonlite_1.8.4                graph_1.74.0                 
 [29] glue_1.6.2                    gtable_0.3.1                 
 [31] zlibbioc_1.42.0               XVector_0.36.0               
 [33] DelayedArray_0.22.0           pheatmap_1.0.12              
 [35] DBI_1.1.3                     Rcpp_1.0.10                  
 [37] htmlTable_2.4.1               xtable_1.8-4                 
 [39] progress_1.2.2                foreign_0.8-84               
 [41] bit_4.0.5                     htmlwidgets_1.6.1            
 [43] httr_1.4.5                    ellipsis_0.3.2               
 [45] farver_2.1.1                  pkgconfig_2.0.3              
 [47] XML_3.99-0.13                 nnet_7.3-18                  
 [49] deldir_1.0-6                  sass_0.4.5                   
 [51] locfit_1.5-9.7                utf8_1.2.3                   
 [53] labeling_0.4.2                tidyselect_1.2.0             
 [55] rlang_1.0.6                   later_1.3.0                  
 [57] munsell_0.5.0                 BiocVersion_3.15.2           
 [59] tools_4.2.2                   cachem_1.0.7                 
 [61] cli_3.6.0                     generics_0.1.3               
 [63] RSQLite_2.3.0                 evaluate_0.20                
 [65] fastmap_1.1.1                 yaml_2.3.7                   
 [67] processx_3.8.0                knitr_1.42                   
 [69] bit64_4.0.5                   fs_1.6.1                     
 [71] KEGGREST_1.36.3               RBGL_1.72.0                  
 [73] whisker_0.4.1                 mime_0.12                    
 [75] xml2_1.3.3                    compiler_4.2.2               
 [77] rstudioapi_0.14               filelock_1.0.2               
 [79] curl_5.0.0                    png_0.1-8                    
 [81] interactiveDisplayBase_1.34.0 statmod_1.5.0                
 [83] bslib_0.4.2                   stringi_1.7.12               
 [85] highr_0.10                    ps_1.7.2                     
 [87] RSpectra_0.16-1               Matrix_1.5-3                 
 [89] vctrs_0.5.2                   pillar_1.8.1                 
 [91] lifecycle_1.0.3               BiocManager_1.30.20          
 [93] jquerylib_0.1.4               data.table_1.14.8            
 [95] bitops_1.0-7                  corpcor_1.6.10               
 [97] httpuv_1.6.9                  rtracklayer_1.56.1           
 [99] latticeExtra_0.6-30           R6_2.5.1                     
[101] BiocIO_1.6.0                  promises_1.2.0.1             
[103] codetools_0.2-19              SummarizedExperiment_1.26.1  
[105] rprojroot_2.0.3               rjson_0.2.21                 
[107] withr_2.5.0                   GenomicAlignments_1.32.1     
[109] Rsamtools_2.12.0              GenomeInfoDbData_1.2.8       
[111] parallel_4.2.2                hms_1.1.2                    
[113] rpart_4.1.19                  grid_4.2.2                   
[115] rmarkdown_2.20                MatrixGenerics_1.8.1         
[117] git2r_0.31.0                  getPass_0.2-2                
[119] base64enc_0.1-3               shiny_1.7.4                  
[121] ellipse_0.4.3                 interp_1.1-3                 
[123] restfulr_0.0.15