applying cNMF to multiple datasets?

[erzakiev]

How would you recommend to proceed with multiple samples? In the original manuscript, you conclude that it is better to perform any kind of batch correction before applying cNMF, correct?

[Prathyusha-konda]

Hi, I have a related question - how to proceed with integrated and subsetted data from multiple samples? Thanks! :)

[dylkot]

Hi, sorry for slow response @erzakiev. Yes, that is right, we correct the data before cNMF currently. We have developed an approach recently to integrate multiple samples ahead of time without losing the non-negativity of the data and maintaining similar normalization. It is pretty heuristic but seems to be working well for us when batch is a problem. Hopefully we will publish an analysis using this in the not so distant future. We use harmony but with a hack to correct the data directly rather than the PCA. If you want, shoot me an e-mail at [email protected] and I can send you example code with the approach

[erzakiev]

Thanks for the response, Dylan!! I dropped you a message on gmail.

I tried to do this myself by calculating spectra for individual samples, following the instructions for handling individual samples, and then I hierarchically clustered the resulting spectra from each sample based on their cosine similarity; I then chose a cutoff for the tree to be cut into (see the red line in the plot above), and for the resulting clusters of spectra I just averaged values for each gene and sorted the resulting values in decreasing order, giving me a ranked set of genes that I took only top 50 of to form signatures.

This makeshift approach has provided me with some interesting signatures, some of whom are corroborated by other methods, and also the pathway enrichments are spot-on.

I'll certainly try out the example code you're going to send me, but in the meantime I wonder if that was an unreasonable approach?

[AAAapollo]

I have same question about multi-dataset. And I used SCTransform to remove batch effect. Could I directly use SCT-assay-counts matrix as input?

[dylkot]

Just an update on this topic that the method to batch correct the data prior to cNMF is in the development branch at this time with an example of its use in the Tutorials folder. Give it a try if you are interested.

[Christopher-Dall]

Hi, wanted to follow up on this. I have multiple samples (treated, untreated). Looked in the tutorial folder for this method to batch correct but didn't see anything. Any way to post the harmony pre-processing for combining datasets or another way to correct data prior to cNMF? Thank you in advance!

[dylkot]

Hey Christopher, are you looking in the development branch? GitHub - dylkot/cNMF at developmenthttps://github.com/dylkot/cNMF/tree/development

Cheers, Dylan

From: Christopher-Dall @.> Date: Tuesday, November 14, 2023 at 6:11 PM To: dylkot/cNMF @.> Cc: Kotliar, Dylan A @.>, Comment @.> Subject: Re: [dylkot/cNMF] applying cNMF to multiple datasets? (Issue #51)

Hi, wanted to follow up on this. I have multiple samples (treated, untreated). Looked in the tutorial folder for this method to batch correct but didn't see anything. Any way to post the harmony pre-processing for combining datasets or another way to correct data prior to cNMF? Thank you in advance!

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[Christopher-Dall]

That'll do it! Thank you!!

[dylkot]

Awesome. We plan to push this to the main branch soon. Cheers, Dylan

From: Christopher-Dall @.> Date: Wednesday, November 15, 2023 at 7:13 AM To: dylkot/cNMF @.> Cc: Kotliar, Dylan A @.>, Comment @.> Subject: Re: [dylkot/cNMF] applying cNMF to multiple datasets? (Issue #51)

That'll do it! Thank you!!

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